WO2020069214A2

WO2020069214A2 - Optimized expression systems for producing cannabinoid synthase polypeptides, cannabinoids, and cannabinoid derivatives

Info

Publication number: WO2020069214A2
Application number: PCT/US2019/053292
Authority: WO
Inventors: Andrew HORWITZ; Leo D'ESPAUX; Jeff Wong; Rathin BECTOR; Anna K. HJELMELAND; Darren Platt; Jeff UBERSAX
Original assignee: Demetrix, Inc.
Priority date: 2018-09-26
Filing date: 2019-09-26
Publication date: 2020-04-02
Also published as: WO2020069214A3

Abstract

The present disclosure provides modified host cells that produce a cannabinoid or a cannabinoid derivative and modified host cells that express a cannabinoid synthase polypeptide. The present disclosure also provides methods of synthesizing a cannabinoid or a cannabinoid derivative. Further, the present disclosure provides novel cannabinoids and cannabinoid derivatives, compositions, including pharmaceutical compositions, as well as methods, uses, compounds for use, medicaments, and compositions (including pharmaceutical compositions) for use comprising said cannabinoid and cannabinoid derivatives.

Description

OPTIMIZED EXPRESSION SYSTEMS FOR PRODUCING CANNABINOID SYNTHASE POLYPEPTIDES, CANNABINOIDS, AND CANNABINOID DERIVATIVES

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Application No.

62/851,556, filed May 22, 2019, U.S. Provisional Application No. 62/750, 178, filed October 24, 2018, and U.S. Provisional Application No. 62/736,997, filed September 26, 2018, the contents of each of which are incorporated herein by reference in their entirety.

INTRODUCTION

[0002] Plants from the genus Cannabis have been used by humans for their medicinal properties for thousands of years. In modem times, the bioactive effects of Cannabis are attributed to a class of compounds termed“cannabinoids,” of which there are hundreds of structural analogs including tetrahydrocannabinol (THC) and cannabidiol (CBD). These molecules and preparations of Cannabis material have recently found application as therapeutics for chronic pain, multiple sclerosis, cancer-associated nausea and vomiting, weight loss, appetite loss, spasticity, seizures, and other conditions.

Cannabidiol / CBD Tetrahydrocannabinol / THC / Dronabinol / Marinol

[0003] The physiological effects of certain cannabinoids are thought to be mediated by their interaction with two cellular receptors found in humans and other animals.

Cannabinoid receptor type 1 (CB 1) is common in the brain, the reproductive system, and the eye. Cannabinoid receptor type 2 (CB2) is common in the immune system and mediates therapeutic effects related to inflammation in animal models. The discovery of cannabinoid receptors and their interactions with plant-derived cannabinoids predated the identification of endogenous ligands.

[0004] Besides THC and CBD, hundreds of other cannabinoids have been identified in Cannabis. However, many of these compounds exist at low levels and alongside more abundant cannabinoids, making it difficult to obtain pure samples from plants to study their therapeutic potential. Similarly, methods of chemically synthesizing these types of products have been cumbersome and costly, and tend to produce insufficient yield. Accordingly, additional methods of making pure cannabinoids or cannabinoid derivatives are needed.

[0005] One possible method is production via fermentation of engineered microbes, such as yeast. By engineering production of the relevant plant enzymes in microbes, it may be possible to achieve conversion of various feedstocks into a range of cannabinoids, potentially at much lower cost and with much higher purity than what is available from the plant. A key challenge to this effort is the difficulty of expressing plant enzymes in the microbe, particularly secreted enzymes such as the cannabinoid synthases, which must successfully traverse the microbe’s secretory pathway to fold and function properly. Specific challenges for processing heterologous secretory proteins in microbes may include 1) identification of plant enzyme secretory sequences which may require replacement with analogous microbe sequences, 2) optimization of codon usage to promote efficient translation, 3) support of protein folding and disulfide formation requirements, 4) support of co-factor requirements (e.g., covalently linked flavin adenine dinucleotide (FAD)), and 5) potential for microbe glycosylation patterns to interfere with protein folding or function. New methods and modified microbes are needed to address these challenges.

SUMMARY

[0006] The present disclosure provides modified host cells comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide. The modified host cells of the disclosure may be useful for producing cannabinoids or cannabinoid derivatives (e.g., non-naturally occurring cannabinoids) and/or for expressing a cannabinoid synthase polypeptide. The present disclosure also provides methods, nucleic acids, and modified host cells for the production of cannabinoids or cannabinoid derivatives. Additionally, the present disclosure provides methods, nucleic acids, and modified host cells for the expression of cannabinoid synthase polypeptides. Further, the present disclosure provides novel cannabinoids and cannabinoid derivatives, compositions, including pharmaceutical compositions, as well as methods, uses, compounds for use, medicaments, and compositions (including pharmaceutical compositions) for use comprising said cannabinoid and cannabinoid derivatives

[0007] One aspect of the disclosure relates to a nucleic acid comprising a codon- optimized nucleotide sequence having at least 85% sequence identity to SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8. In certain such embodiments, the codon-optimized nucleotide sequence encodes a cannabinoid synthase polypeptide. In certain such embodiments, the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[0008] Another aspect of the disclosure relates to a nucleic acid comprising a codon- optimized nucleotide sequence having at least 85% sequence identity to SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO:263.

In certain such embodiments, the codon-optimized nucleotide sequence encodes a cannabinoid synthase polypeptide. In certain such embodiments, the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[0009] Another aspect of the disclosure relates to a nucleic acid comprising a codon- optimized nucleotide sequence set forth in SEQ ID NO:389, SEQ ID NO:39l, SEQ ID NO:393, SEQ ID NO:395, SEQ ID N0 397, SEQ ID N0 399, SEQ ID NO:40l, SEQ ID NO: 403, SEQ ID NO: 405, SEQ ID NO: 407, SEQ ID NO: 409, SEQ ID NO:4l 1, SEQ ID NO:4l3, SEQ ID NO:4l5, SEQ ID NO:4l7, SEQ ID NO:4l9, SEQ ID NO:42l, SEQ ID NO: 423, SEQ ID NO: 425, SEQ ID NO 427, SEQ ID NO: 429, SEQ ID NO: 431, SEQ ID NO:433, SEQ ID NO:435, SEQ ID NO:437, SEQ ID N0 439, SEQ ID NO:44l, SEQ ID NO: 443, SEQ ID NO: 445, SEQ ID NO: 447, SEQ ID NO: 449, SEQ ID NO: 451, SEQ ID NO:453, SEQ ID NO:455, SEQ ID N0 457, SEQ ID N0 459, SEQ ID NO:46l, SEQ ID NO: 463, SEQ ID NO: 465, SEQ ID NO: 467, SEQ ID NO: 469, SEQ ID NO: 471, SEQ ID NO: 473, SEQ ID NO: 475, SEQ ID NO: 477, SEQ ID NO: 479, SEQ ID NO: 481, SEQ ID NO:483, SEQ ID NO:485, SEQ ID NO:487, SEQ ID N0 489, SEQ ID NO:49l, SEQ ID NO:493, SEQ ID NO:495, SEQ ID NO:497, SEQ ID NO:499, SEQ ID NO:50l, SEQ ID NO:503, SEQ ID NO: 505, SEQ ID NO:507, SEQ ID NO:509, SEQ ID NO:5l l, SEQ ID N0 513, SEQ ID NO: 515, SEQ ID N0 517, SEQ ID NO 519, SEQ ID N0 521, SEQ ID NO:523, SEQ ID NO:525, SEQ ID NO:527, SEQ ID NO:529, SEQ ID NO:53 l, SEQ ID NO:533, SEQ ID NO:535, SEQ ID NO:537, SEQ ID N0 539, SEQ ID NO:54l, SEQ ID NO:543, SEQ ID NO:545, SEQ ID N0 547, SEQ ID N0 549, SEQ ID NO:55l, SEQ ID NO:553, SEQ ID NO:555, SEQ ID NO:557, SEQ ID N0 559, SEQ ID NO:56l, SEQ ID NO:563, SEQ ID NO:565, SEQ ID NO:567, SEQ ID N0 569, SEQ ID NO:57l, SEQ ID NO:573, SEQ ID NO:579, SEQ ID N0 581, SEQ ID N0 583, SEQ ID NO:585, SEQ ID NO:587, SEQ ID NO:589, SEQ ID NO:59l, SEQ ID N0 593, SEQ ID NO:595, SEQ ID NO:597, or SEQ ID NO:599. In certain such embodiments, the codon-optimized nucleotide sequence encodes a cannabinoid synthase polypeptide. In certain such embodiments, the cannabinoid synthase polypeptide is a variant cannabidiolic acid synthase polypeptide. [0010] Another aspect of the disclosure relates to a nucleic acid comprising a codon- optimized nucleotide sequence set forth in SEQ ID NO:606, SEQ ID NO:608, SEQ ID NO:6lO, SEQ ID NO:6l2, SEQ ID NO:6l4, SEQ ID NO:6l6, SEQ ID NO:6l8, SEQ ID NO:620, SEQ ID NO:622, SEQ ID NO:624, SEQ ID N0 626, SEQ ID NO:628, SEQ ID

NO:630, SEQ ID NO:632, SEQ ID NO:634, SEQ ID N0 636, SEQ ID NO:638, SEQ ID

NO:640, SEQ ID NO:642, SEQ ID NO:644, SEQ ID NO:646, SEQ ID NO:648, SEQ ID NO:650, SEQ ID NO:652, SEQ ID NO:654, SEQ ID N0 656, SEQ ID NO:658, SEQ ID

NO:660, SEQ ID NO:662, SEQ ID NO:664, SEQ ID N0 666, SEQ ID NO:668, SEQ ID

NO:670, SEQ ID NO:672, SEQ ID NO:674, SEQ ID N0 676, SEQ ID NO:678, SEQ ID

NO:680, SEQ ID NO:682, SEQ ID NO:684, SEQ ID N0 686, SEQ ID NO:688, SEQ ID

NO:690, SEQ ID NO:692, SEQ ID N0 694, SEQ ID N0 696, SEQ ID NO:698, SEQ ID

NO:700, SEQ ID NO:702, SEQ ID NO:704, SEQ ID NO:706, SEQ ID NO:708, SEQ ID NO:7lO, SEQ ID NO:7l2, SEQ ID NO:7l4, SEQ ID NO:7l6, SEQ ID NO:7l8, SEQ ID NO:720, SEQ ID NO:722, SEQ ID N0 724, SEQ ID N0 726, SEQ ID NO:728, SEQ ID

NO:730, SEQ ID NO:732, SEQ ID NO:734, SEQ ID N0 736, SEQ ID NO:738, SEQ ID

NO:740, or SEQ ID NO:742. In certain such embodiments, the codon-optimized nucleotide sequence encodes a cannabinoid synthase polypeptide. In certain such embodiments, the cannabinoid synthase polypeptide is a variant tetrahydrocannabinolic acid synthase polypeptide.

[0011] One aspect of the disclosure relates to a nucleic acid comprising a codon- optimized nucleotide sequence having at least 85% sequence identity to SEQ ID NO:238, SEQ ID NO:27l, SEQ ID NO:272, SEQ ID NO:273, or SEQ ID NO:274. In certain such embodiments, the codon-optimized nucleotide sequence encodes a cannabinoid synthase polypeptide. In certain such embodiments, the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide.

[0012] Another aspect of the disclosure relates to a nucleic acid comprising a codon- optimized nucleotide sequence having at least 85% sequence identity to SEQ ID NO:240. In certain such embodiments, the codon-optimized nucleotide sequence encodes a cannabinoid synthase polypeptide. In certain such embodiments, the cannabinoid synthase polypeptide is a cannabichromenic acid synthase polypeptide.

[0013] One aspect of the disclosure relates to a nucleic acid comprising a codon- optimized nucleotide sequence having at least 85% sequence identity to SEQ ID NO:244. In certain such embodiments, the codon-optimized nucleotide sequence encodes a cannabinoid synthase polypeptide. In certain such embodiments, the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[0014] In some embodiments described herein, the nucleic acid comprises a nucleotide sequence encoding a signal sequence polypeptide. In some embodiments, the signal sequence polypeptide is a secretory signal sequence polypeptide. In some embodiments, the secretory signal sequence polypeptide is a native secretory signal sequence polypeptide. In some embodiments, the secretory signal sequence polypeptide is a synthetic secretory signal sequence polypeptide. In some embodiments, the secretory signal sequence polypeptide is an endoplasmic reticulum retention signal sequence polypeptide. In some embodiments, the secretory signal sequence polypeptide is a vacuolar localization signal sequence polypeptide. In certain such embodiments, the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide or a PRClt polypeptide. In certain such embodiments, the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide. In some embodiments, the secretory signal sequence polypeptide is a plasma membrane localization signal sequence polypeptide. In some embodiments, the secretory signal sequence polypeptide is a peroxisome targeting signal sequence polypeptide.

[0015] One aspect of the disclosure relates to a method of making a modified host cell for producing a cannabinoid or cannabinoid derivative, the method comprising introducing one or more nucleic acids described herein into a host cell.

[0016] Another aspect of the disclosure relates to a vector comprising one or more nucleic acids described herein.

[0017] One aspect of the disclosure relates to a method of making a modified host cell for producing a cannabinoid or a cannabinoid derivative, the method comprising introducing one or more vectors described herein into a host cell.

[0018] Another aspect of the disclosure relates to a modified host cell for producing a cannabinoid or cannabinoid derivative comprising one or more nucleic acids described herein. In some embodiments, the modified host cell is a eukaryotic cell. In certain such embodiments, the eukaryotic cell is a yeast cell. In certain such embodiments, the yeast cell is Saccharomyces cerevisiae. In certain such embodiments, the Saccharomyces cerevisiae is a protease-deficient strain of Saccharomyces cerevisiae. In some embodiments described herein, the one or more nucleic acids are integrated into the chromosome of the modified host cell. In some embodiments described herein, at least one of the one or more nucleic acids are maintained extrachromosomally (e.g., on a plasmid or artificial chromosome). In some embodiments described herein, at least one of the one or more nucleic acids are operably-linked to an inducible promoter. In some embodiments described herein, at least one of the one or more nucleic acids are operably-linked to a constitutive promoter.

[0019] One aspect of the disclosure relates to a modified host cell for producing a cannabinoid or cannabinoid derivative, wherein the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides or a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, or a combination of any of the foregoing, and wherein the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide. In certain such embodiments, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides. In certain such embodiments, the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin mononucleotide (FMN) synthetase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosidase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosyltransferase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more peptidyl-prolyl isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more fatty acid desaturase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein transport and trafficking polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more signal peptidase or signal peptidase complex polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR), one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in endoplasmic reticulum-associated degradation (ERAD), one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cell wall assembly, or one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in vacuolar protein sorting, or a combination of any of the foregoing. In some embodiments, the nucleotide sequences encoding the one or more secretory pathway polypeptides are codon- optimized.

[0020] In some embodiments described herein, the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide, a !EMl polypeptide, a LHSl polypeptide, a SIS1 polypeptide, a SSB1 polypeptide, a CNE1 polypeptide, a CNS1 polypeptide, a PFD2s polypeptide, a PFDl polypeptide, a SSA1 polypeptide, a YDJ1 polypeptide, a SIL1 polypeptide, a SCJ1 polypeptide, a ROT1 polypeptide, a TCP1 polypeptide, a CCT2 polypeptide, a CCT3 polypeptide, a CCT4 polypeptide, a CCT5 polypeptide, a CCT6 polypeptide, a CCT7 polypeptide, a CCT8 polypeptide, a DEDl polypeptide, a CPR5 polypeptide, or a FPRl polypeptide, or a combination of any of the foregoing. In certain such embodiments, the modified host cell comprises two or more heterologous nucleic acids comprising a nucleotide sequence encoding a KAR2 polypeptide. In certain such embodiments, the modified host cell comprises three or more heterologous nucleic acids comprising a nucleotide sequence encoding a KAR2 polypeptide.

[0021] In some embodiments described herein, the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response. In certain such embodiments, the one or more polypeptides involved in unfolded protein response comprise an IRE1 polypeptide, a HACls polypeptide, a DED1 polypeptide, or a PPQ1 polypeptide, or a combination of any of the foregoing. In some embodiments described herein, the one or more polypeptides involved in unfolded protein response comprise a transcription factor polypeptide or a lumenal sensor polypeptide that binds to an unfolded protein response element. In some embodiments described herein, the modified host cell comprises a synthetic polypeptide that binds to an unfolded protein response element.

[0022] In some embodiments described herein, the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum. In certain such embodiments, the one or more polypeptides involved in protein translocation into the endoplasmic reticulum comprise a KAR2 polypeptide, a SEC61 polypeptide, a SBH1 polypeptide, a SSS1 polypeptide, a SRP14 polypeptide, a SRP21 polypeptide, a SRP68 polypeptide, a SRP72 polypeptide, a SEC65 polypeptide, a SRP54 polypeptide, a SEC62 polypeptide, a SEC63 polypeptide, a SEC66 polypeptide, a SEC72 polypeptide, a SRP101 polypeptide, or a SRP102 polypeptide, or a combination of any of the foregoing. In some embodiments described herein, the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding SRP protein complex RNA component SCR1.

[0023] In some embodiments described herein, the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides. In certain such embodiments, the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FADl polypeptide.

[0024] In some embodiments described herein, the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin mononucleotide (FMN) synthetase polypeptides. In certain such embodiments, the one or more flavin mononucleotide (FMN) synthetase polypeptides comprise a FMNl polypeptide.

[0025] In some embodiments described herein, the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosidase polypeptides. In certain such embodiments, the one or more glycosidase polypeptides comprise a CWH41 polypeptide.

[0026] In some embodiments described herein, the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more fatty acid desaturase polypeptides. In certain such embodiments, the one or more fatty acid desaturase polypeptides comprise an OLE1 polypeptide.

[0027] In some embodiments described herein, the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides. In certain such embodiments, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide.

[0028] In some embodiments described herein, the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides. In certain such embodiments, the one or more thiol oxidase polypeptides comprise an EROl polypeptide or an ERV2 polypeptide, or a combination of any of the foregoing. In certain such embodiments, the modified host cell comprises two or more heterologous nucleic acids comprising a nucleotide sequence encoding an EROl polypeptide.

[0029] In some embodiments described herein, the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more signal peptidase or signal peptidase complex polypeptides. In certain such embodiments, the one or more signal peptidase or signal peptidase complex polypeptides comprise a KEX2 polypeptide, a SPC1 polypeptide, a SPC2 polypeptide, a SPC3 polypeptide, or an SEC11 polypeptide, or a combination of any of the foregoing.

[0030] In some embodiments described herein, the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein transport and trafficking polypeptides. In certain such embodiments, the one or more protein transport and trafficking polypeptides comprise a SEC23 polypeptide, a SEC24 polypeptide, a SEC 13 polypeptide, a BFR2 polypeptide, a SEC31 polypeptide, a SAR1 polypeptide, a SEC12 polypeptide, a SEC4 polypeptide, a SEC 16 polypeptide, an ERV29 polypeptide, a SEC7 polypeptide, a SEC5 polypeptide, a SEC22 polypeptide, a BOSl polypeptide, a BETl polypeptide, a BElGl polypeptide, a GRH1 polypeptide, a USOl polypeptide, a SEC17 polypeptide, a SEC18 polypeptide, a SSOl polypeptide, a SEC9 polypeptide, a SNC2 polypeptide, or a LDB17 polypeptide, or a combination of any of the foregoing.

[0031] In some embodiments described herein, the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in endoplasmic reticulum-associated degradation (ERAD). In certain such embodiments, the one or more polypeptides involved in endoplasmic reticulum-associated degradation (ERAD) comprise a PL4 polypeptide.

[0032] In some embodiments described herein, the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cell wall assembly. In certain such embodiments, the one or more polypeptides involved in cell wall assembly comprise a KRE1 polypeptide.

[0033] In some embodiments described herein, the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in vacuolar protein sorting. In certain such embodiments, the one or more polypeptides involved in vacuolar protein sorting comprise a PEPl polypeptide.

[0034] In some embodiments described herein, the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosyltransferase polypeptides.

[0035] In some embodiments described herein, the modified host cell comprises a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides. In certain such embodiments, the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more glycosidase polypeptides, one or more genes encoding one or more glycosyltransferase polypeptides, one or more genes encoding one or more polypeptides involved in lipid droplet assembly, one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism, or one or more genes encoding one or more vacuolar proteinase polypeptides, or a combination of any of the foregoing. [0036] In some embodiments described herein, the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more glycosidase polypeptides. In certain such embodiments, the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene or a MNS1 gene, or a combination of any of the foregoing.

[0037] In some embodiments described herein, the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more glycosyltransferase polypeptides. In certain such embodiments, the one or more genes encoding one or more glycosyltransferase polypeptides comprise an ALG12 gene.

[0038] In some embodiments described herein, the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more polypeptides involved in lipid droplet assembly. In certain such embodiments, the one or more genes encoding one or more polypeptides involved in lipid droplet assembly comprise a FLDl gene.

[0039] In some embodiments described herein, the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism. In certain such embodiments, the one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism comprise an OPI1 gene.

[0040] In some embodiments described herein, the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides. In certain such embodiments, the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene, a PRB1 gene, or a PRC1 gene, or a combination of any of the foregoing.

[0041] In some embodiments described herein, the modified host cell comprises two or more heterologous nucleic acids comprising a nucleotide sequence encoding the cannabinoid synthase polypeptide.

[0042] In some embodiments described herein, the nucleotide sequence encoding the cannabinoid synthase polypeptide is codon-optimized.

[0043] In some embodiments described herein, the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. In some embodiments described herein, the cannabinoid synthase polypeptide is a cannabichromenic acid synthase polypeptide. In some embodiments described herein, the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[0044] In some embodiments, the cannabinoid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 85% sequence identity to SEQ ID NO:242 or SEQ ID NO:244. In some embodiments, the cannabidiolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence set forth in SEQ ID NO: 242 or SEQ ID NO: 244.

[0045] In some embodiments described herein, the cannabidiolic acid synthase polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:243 or SEQ ID NO:245.

[0046] In some embodiments described herein, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide. In some embodiments, the signal sequence polypeptide is a secretory signal sequence polypeptide. In some embodiments, the secretory signal sequence polypeptide is a native secretory signal sequence polypeptide. In some embodiments, the secretory signal sequence polypeptide is a synthetic secretory signal sequence polypeptide. In some embodiments, the secretory signal sequence polypeptide is an endoplasmic reticulum retention signal sequence polypeptide. In some embodiments, the secretory signal sequence polypeptide is a vacuolar localization signal sequence polypeptide. In certain such embodiments, the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide or a PRClt polypeptide. In certain such embodiments, the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide. In some embodiments, the secretory signal sequence polypeptide is a plasma membrane localization signal sequence polypeptide. In some embodiments, the secretory signal sequence polypeptide is a peroxisome targeting signal sequence polypeptide.

[0047] In some embodiments of the modified host cell of the disclosure, the one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide comprise a nucleotide sequence encoding a signal sequence polypeptide.

[0048] In some embodiments described herein, the cannabidiolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 85% sequence identity to SEQ ID NO: l, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, or SEQ ID NO: 8. In some embodiments, the cannabidiolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence set forth in SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8.

[0049] In some embodiments described herein, the cannabidiolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 85% sequence identity to SEQ ID NO:256, SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO:263. In some embodiments, the cannabidiolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence set forth in SEQ ID NO:256, SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260,

SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO:263.

[0050] In some embodiments described herein, the cannabidiolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence set forth in SEQ ID

NO: 389, SEQ ID NO 391, SEQ ID NO:393, SEQ ID NO: 395, SEQ ID NO: 397, SEQ ID NO: 399, SEQ ID NO 401, SEQ ID NO:403, SEQ ID NO 405, SEQ ID NO: 407, SEQ ID NO: 409, SEQ ID NO 411, SEQ ID N0 413, SEQ ID N0 415, SEQ ID NO:4l7, SEQ ID NO:4l9, SEQ ID NO 421, SEQ ID NO:423, SEQ ID N0 425, SEQ ID NO: 427, SEQ ID NO: 429, SEQ ID NO 431, SEQ ID NO:433, SEQ ID N0 435, SEQ ID NO: 437, SEQ ID NO: 439, SEQ ID NO 441, SEQ ID N0 443, SEQ ID N0 445, SEQ ID NO: 447, SEQ ID NO: 449, SEQ ID NO 451, SEQ ID NO:453, SEQ ID N0 455, SEQ ID NO: 457, SEQ ID NO:459, SEQ ID NO 461, SEQ ID NO:463, SEQ ID N0 465, SEQ ID NO: 467, SEQ ID NO: 469, SEQ ID NO 471, SEQ ID N0 473, SEQ ID NO 475, SEQ ID NO: 477, SEQ ID NO: 479, SEQ ID NO 481, SEQ ID NO:483, SEQ ID N0 485, SEQ ID NO:487, SEQ ID NO:489, SEQ ID NO 491, SEQ ID NO:493, SEQ ID N0 495, SEQ ID NO: 497, SEQ ID NO: 499, SEQ ID NO 501, SEQ ID NO 503, SEQ ID NO 505, SEQ ID NO: 507, SEQ ID NO: 509, SEQ ID NO 511, SEQ ID NO:5 l3, SEQ ED N0 515, SEQ ID NO:5l7, SEQ ID NO:5l9, SEQ ID NO 521, SEQ ID NO:523, SEQ ID NO 525, SEQ ID NO: 527, SEQ ID NO: 529, SEQ ID NO 531, SEQ ID N0 533, SEQ ID N0 535, SEQ ID NO: 537, SEQ ID NO: 539, SEQ ID NO 541, SEQ ID NO:543, SEQ ID NO 545, SEQ ID NO: 547, SEQ ID NO: 549, SEQ ID NO 551, SEQ ID NO:553, SEQ ID N0 555, SEQ ID NO:557, SEQ ID NO:559, SEQ ID NO 561, SEQ ID N0 563, SEQ ID N0 565, SEQ ID NO: 567, SEQ ID NO: 569, SEQ ID NO 571, SEQ ID NO:573, SEQ ID NO 579, SEQ ID NO:58l, SEQ ID NO:583, SEQ ID NO 585, SEQ ID NO: 587, SEQ ID NO: 589, SEQ ID NO:59l, SEQ ID NO:593, SEQ ID NO 595, SEQ ID NO: 597, or SEQ ID NO:599. In certain such embodiments, the cannabidiolic acid synthase polypeptide is a variant cannabidiolic acid synthase polypeptide. [0051] In some embodiments described herein, the tetrahydrocannabinolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence set forth in SEQ ID NO:606, SEQ ID NO:608, SEQ ID NO:6lO, SEQ ID NO:6l2, SEQ ID NO:6l4, SEQ ID NO:6l6, SEQ ID NO:6l8, SEQ ID NO:620, SEQ ID N0 622, SEQ ID NO:624, SEQ ID NO:626, SEQ ID NO:628, SEQ ID NO:630, SEQ ID N0 632, SEQ ID NO:634, SEQ ID NO:636, SEQ ID NO:638, SEQ ID NO:640, SEQ ID N0 642, SEQ ID NO:644, SEQ ID NO:646, SEQ ID NO:648, SEQ ID NO:650, SEQ ID N0 652, SEQ ID NO:654, SEQ ID NO:656, SEQ ID NO:658, SEQ ID NO:660, SEQ ID N0 662, SEQ ID NO:664, SEQ ID NO:666, SEQ ID NO:668, SEQ ID NO:670, SEQ ID N0 672, SEQ ID NO:674, SEQ ID NO:676, SEQ ID NO:678, SEQ ID NO:680, SEQ ID N0 682, SEQ ID NO:684, SEQ ID NO:686, SEQ ID NO:688, SEQ ID NO 690, SEQ ID N0 692, SEQ ID NO:694, SEQ ID NO:696, SEQ ID NO:698, SEQ ID N0:700, SEQ ID NO:702, SEQ ID NO:704, SEQ ID NO:706, SEQ ID NO:708, SEQ ID NO:7lO, SEQ ID NO:7l2, SEQ ID NO:7l4, SEQ ID NO:7l6, SEQ ID NO:7l8, SEQ ID NO 720, SEQ ID N0 722, SEQ ID NO:724, SEQ ID NO:726, SEQ ID NO:728, SEQ ID NO:730, SEQ ID N0 732, SEQ ID NO:734, SEQ ID NO:736, SEQ ID NO:738, SEQ ID NO:740, or SEQ ID NO:742. In certain such embodiments, the tetrahydrocannabinolic acid synthase polypeptide is a variant tetrahydrocannabinolic acid synthase polypeptide.

[0052] In some embodiments described herein, the tetrahydrocannabinolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 85% sequence identity to SEQ ID NO:238, SEQ ID NO:271, SEQ ID NO:272, SEQ ID NO:273, or SEQ ID NO:274. In some embodiments, the tetrahydrocannabinolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence set forth in SEQ ID NO:238, SEQ ID NO:271, SEQ ID NO:272, SEQ ID NO:273, or SEQ ID NO:274.

[0053] In some embodiments described herein, the tetrahydrocannabinolic acid synthase polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:239.

[0054] In some embodiments described herein, the tetrahydrocannabinolic acid synthase polypeptide is a variant tetrahydrocannabinolic acid synthase polypeptide comprising an amino acid sequence set forth in SEQ ID NO:607, SEQ ID NO:609, SEQ ID NO:6l l, SEQ ID NO:6l3, SEQ ID NO:6l5, SEQ ID NO:6l7, SEQ ID NO:6l9, SEQ ID NO:62l, SEQ ID NO:623, SEQ ID NO:625, SEQ ID N0 627, SEQ ID NO:629, SEQ ID NO:63 l, SEQ ID NO:633, SEQ ID NO:635, SEQ ID NO:637, SEQ ID NO:639, SEQ ID NO:64l, SEQ ID NO:643, SEQ ID NO:645, SEQ ID N0 647, SEQ ID NO:649, SEQ ID NO:65l, SEQ ID NO:653, SEQ ID NO:655, SEQ ID N0 657, SEQ ID NO:659, SEQ ID NO:66l, SEQ ID NO:663, SEQ ID NO:665, SEQ ID N0 667, SEQ ID NO:669, SEQ ID

NO:67l, SEQ ID NO:673, SEQ ID NO:675, SEQ ID N0 677, SEQ ID NO:679, SEQ ID

NO:68l, SEQ ID NO:683, SEQ ID NO:685, SEQ ID N0 687, SEQ ID NO:689, SEQ ID

NO:69l, SEQ ID NO:693, SEQ ID NO:695, SEQ ID N0 697, SEQ ID NO:699, SEQ ID

NO:70l, SEQ ID NO:703, SEQ ID NO:705, SEQ ID NO:707, SEQ ID NO:709, SEQ ID N0:7l l, SEQ ID NO:7l3, SEQ ID NO:7l5, SEQ ID N0 717, SEQ ID NO:7l9, SEQ ID NO:72l, SEQ ID NO:723, SEQ ID NO:725, SEQ ID N0 727, SEQ ID NO:729, SEQ ID NO:73 l, SEQ ID NO:733, SEQ ID NO:735, SEQ ID N0 737, SEQ ID NO:739, SEQ ID NO:74l, or SEQ ID NO:743.

[0055] In some embodiments described herein, the cannabichromenic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 85% sequence identity to SEQ ID NO:240. In some embodiments, the cannabichromenic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence set forth in SEQ ID NO: 240.

[0056] In some embodiments described herein, the cannabichromenic acid synthase polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:241.

[0057] In some embodiments described herein, the cannabidiolic acid synthase polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:9. In some embodiments described herein, the cannabidiolic acid synthase polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID N0 257

[0058] In some embodiments described herein, the cannabidiolic acid synthase polypeptide is a variant cannabidiolic acid synthase polypeptide comprising an amino acid sequence set forth in SEQ ID NO: 390, SEQ ID NO: 392, SEQ ID NO: 394, SEQ ID NO: 396, SEQ ID NO:398, SEQ ID NO:400, SEQ ID NO:402, SEQ ID NO:404, SEQ ID NO:406,

SEQ ID NO:408, SEQ ID NO:410, SEQ ID NO:412, SEQ ID NO:4l4, SEQ ID NO:4l6,

SEQ ID NO:418, SEQ ID NO:420, SEQ ID NO:422, SEQ ID NO:424, SEQ ID NO:426,

SEQ ID NO:428, SEQ ID NO:430, SEQ ID NO:432, SEQ ID NO:434, SEQ ID NO:436,

SEQ ID NO:438, SEQ ID NO:440, SEQ ID NO:442, SEQ ID NO:444, SEQ ID NO:446,

SEQ ID NO:448, SEQ ID NO:450, SEQ ID NO:452, SEQ ID NO:454, SEQ ID NO:456,

SEQ ID NO:458, SEQ ID NO:460, SEQ ID NO:462, SEQ ID NO:464, SEQ ID NO:466,

SEQ ID NO:468, SEQ ID NO:470, SEQ ID NO:472, SEQ ID NO:474, SEQ ID NO:476, SEQ ID NO:478, SEQ ID NO:480, SEQ ID NO:482, SEQ ID NO:484, SEQ ID NO:486,

SEQ ID NO:488, SEQ ID NO:490, SEQ ID NO:492, SEQ ID NO:494, SEQ ID NO:496,

SEQ ID NO:498, SEQ ID NO:500, SEQ ID NO:502, SEQ ID NO:504, SEQ ID NO:506,

SEQ ID NO:508, SEQ ID NO:510, SEQ ID NO:512, SEQ ID NO:5l4, SEQ ID NO:5l6, SEQ ID NO:518, SEQ ID NO: 520, SEQ ID NO: 522, SEQ ID NO: 524, SEQ ID NO: 526,

SEQ ID NO:528, SEQ ID NO:530, SEQ ID NO:532, SEQ ID NO:534, SEQ ID NO:536,

SEQ ID NO:538, SEQ ID NO:540, SEQ ID NO:542, SEQ ID NO:544, SEQ ID NO:546,

SEQ ID NO:548, SEQ ID NO:550, SEQ ID NO:552, SEQ ID NO:554, SEQ ID NO:556,

SEQ ID NO:558, SEQ ID NO:560, SEQ ID NO:562, SEQ ID NO:564, SEQ ID NO:566,

SEQ ID NO:568, SEQ ID NO:570, SEQ ID NO:572, SEQ ID NO:574, SEQ ID NO:580,

SEQ ID NO:582, SEQ ID NO:584, SEQ ID NO:586, SEQ ID NO:588, SEQ ID NO:590,

SEQ ID NO:592, SEQ ID NO:594, SEQ ID NO:596, SEQ ID NO:598, or SEQ ID NO:600.

[0059] In some embodiments described herein, the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate: olivetolic acid geranyltransferase (GOT) polypeptide comprising an amino acid sequence having at least 85% sequence identity to SEQ ID NO:73 or SEQ ID NO:70.

[0060] In some embodiments described herein, the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a tetraketide synthase (TKS) polypeptide and one or more heterologous nucleic acids comprising a nucleotide sequence encoding an olivetolic acid cyclase (OAC) polypeptide. In certain such embodiments, the TKS polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:75. In certain such embodiments, the modified host cell comprises three or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide. In some embodiments, the OAC polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:77 or SEQ ID NO:225. In certain such embodiments, the modified host cell comprises three or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide.

[0061] In some embodiments described herein, the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acyl activating enzyme (AAE) polypeptide. In certain such embodiments, the AAE polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:79, SEQ ID NO:268, or SEQ ID NQ:270. In some embodiments, the modified host cell comprises two or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide. [0062] In some embodiments described herein, the modified host cell comprises one or more of the following: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMG-CoA synthase (HMGS) polypeptide; b) one or more

heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3- hydroxy-3-methyl-glutaryl-CoA reductase (tHMGR) polypeptide; c) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate kinase (MK) polypeptide; d) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a phosphomevalonate kinase (PMK) polypeptide; e) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate pyrophosphate decarboxylase (MVD1) polypeptide; or f) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a isopentenyl diphosphate isomerase (IDI1) polypeptide. In some embodiments, the IDI1 polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:8l . In some embodiments, the tHMGR polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:83. In some embodiments, the HMGS polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:85. In some embodiments, the MK polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:95. In some embodiments, the PMK polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:93. In some embodiments, the MVD1 polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:89.

[0063] In some embodiments described herein, the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide. In certain such embodiments, the acetoacetyl-CoA thiolase polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:87.

[0064] In some embodiments described herein, the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a pyruvate decarboxylase (PDC) polypeptide. In certain such embodiments, the PDC polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:9l .

[0065] In some embodiments described herein, the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate synthetase (GPPS) polypeptide. In certain such embodiments, the GPPS polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:97.

[0066] In some embodiments, the disclosure relates to a modified host cell for producing a cannabinoid or cannabinoid derivative, wherein the modified host cell comprises:

a) two or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide;

b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMG-CoA synthase (HMGS) polypeptide;

c) two or more heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3 -hydroxy-3 -methyl-glutaryl-CoA reductase (tHMGR) polypeptide;

d) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate kinase (MK) polypeptide;

e) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a phosphomevalonate kinase (PMK) polypeptide;

f) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate pyrophosphate decarboxylase (MVD1) polypeptide;

g) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a isopentenyl diphosphate isomerase (IDI1) polypeptide;

h) one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide;

i) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a pyruvate decarboxylase (PDC) polypeptide;

j) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate synthetase (GPPS) polypeptide;

k) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate:olivetolic acid geranyltransferase (GOT) polypeptide;

l) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise: 1) two or more heterologous nucleic acids comprising a nucleotide sequence encoding a thiol oxidase polypeptide, wherein the thiol oxidase polypeptide is an EROl polypeptide;

2) two or more heterologous nucleic acids comprising a nucleotide sequence encoding a chaperone or co-chaperone polypeptide, wherein the chaperone or co-chaperone polypeptide is a KAR2 polypeptide;

3) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a polypeptide involved in unfolded protein response, wherein the polypeptide involved in unfolded protein response is an IRE1 polypeptide; and

4) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a protein disulfide isomerase polypeptide, wherein the protein disulfide isomerase polypeptide is a PDI1 polypeptide; and

m) deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise:

1) a gene encoding a glycosidase polypeptide, wherein the gene encoding the glycosidase polypeptide is a ROT2 gene; and

2) a gene encoding a vacuolar proteinase polypeptide, wherein the gene encoding the vacuolar proteinase polypeptide is a PEP4 gene.

In certain such embodiments, the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide, one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide, and/or one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acyl-activating enzyme (AAE) polypeptide.

[0067] In some embodiments, the disclosure relates to a modified host cell for producing a cannabinoid or cannabinoid derivative, wherein the modified host cell comprises:

a) two heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide;

b) a heterologous nucleic acid comprising a nucleotide sequence encoding a HMG- CoA synthase (HMGS) polypeptide; c) two heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3-hydroxy-3-methyl-glutaryl-CoA reductase (tHMGR) polypeptide;

d) a heterologous nucleic acid comprising a nucleotide sequence encoding a mevalonate kinase (MK) polypeptide;

e) a heterologous nucleic acid comprising a nucleotide sequence encoding a phosphomevalonate kinase (PMK) polypeptide;

f) a heterologous nucleic acid comprising a nucleotide sequence encoding a mevalonate pyrophosphate decarboxylase (MVD1) polypeptide;

g) a heterologous nucleic acid comprising a nucleotide sequence encoding a isopentenyl diphosphate isomerase (IDI1) polypeptide;

h) a heterologous nucleic acid comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide;

i) a heterologous nucleic acid comprising a nucleotide sequence encoding a pyruvate decarboxylase (PDC) polypeptide;

j) a heterologous nucleic acid comprising a nucleotide sequence encoding a geranyl pyrophosphate synthetase (GPPS) polypeptide;

k) a heterologous nucleic acid comprising a nucleotide sequence encoding a geranyl pyrophosphate:olivetolic acid geranyltransferase (GOT) polypeptide;

l) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise:

1) two heterologous nucleic acids comprising a nucleotide sequence encoding a thiol oxidase polypeptide, wherein the thiol oxidase polypeptide is an EROl polypeptide;

2) two heterologous nucleic acids comprising a nucleotide sequence encoding a chaperone or co-chaperone polypeptide, wherein the chaperone or co chaperone polypeptide is a KAR2 polypeptide;

3) a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in unfolded protein response, wherein the polypeptide involved in unfolded protein response is an IRE1 polypeptide; and

4) a heterologous nucleic acid comprising a nucleotide sequence encoding a protein disulfide isomerase polypeptide, wherein the protein disulfide isomerase polypeptide is a PDIl polypeptide; and m) deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise:

[0068] In some embodiments, the disclosure relates to a modified host cell for producing a cannabinoid or cannabinoid derivative, wherein the modified host cell comprises:

h) one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide; i) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a pyruvate decarboxylase (PDC) polypeptide;

1) two or more heterologous nucleic acids comprising a nucleotide sequence encoding a chaperone or co-chaperone polypeptide, wherein the chaperone or co-chaperone polypeptide is a KAR2 polypeptide;

2) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a polypeptide involved in unfolded protein response, wherein the polypeptide involved in unfolded protein response is an IRE1 polypeptide; and

3) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a protein disulfide isomerase polypeptide, wherein the protein disulfide isomerase polypeptide is a PDI1 polypeptide; and

In certain such embodiments, the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide, one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide, and/or one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acyl-activating enzyme (AAE) polypeptide. [0069] In some embodiments, the disclosure relates to a modified host cell for producing a cannabinoid or cannabinoid derivative, wherein the modified host cell comprises:

b) a heterologous nucleic acid comprising a nucleotide sequence encoding a HMG- CoA synthase (HMGS) polypeptide;

c) two heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3-hydroxy-3-methyl-glutaryl-CoA reductase (tHMGR) polypeptide;

1) two heterologous nucleic acids comprising a nucleotide sequence encoding a chaperone or co-chaperone polypeptide, wherein the chaperone or co- chaperone polypeptide is a KAR2 polypeptide; 2) a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in unfolded protein response, wherein the polypeptide involved in unfolded protein response is an IRE1 polypeptide; and

3) a heterologous nucleic acid comprising a nucleotide sequence encoding a protein disulfide isomerase polypeptide, wherein the protein disulfide isomerase polypeptide is a PDIl polypeptide; and

[0070] In some embodiments, the disclosure relates to a modified host cell for producing a cannabinoid or cannabinoid derivative, wherein the modified host cell comprises:

e) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a phosphomevalonate kinase (PMK) polypeptide; f) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate pyrophosphate decarboxylase (MVD1) polypeptide;

1) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a chaperone or co-chaperone polypeptide, wherein the chaperone or co-chaperone polypeptide is a KAR2 polypeptide; and

2) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a protein disulfide isomerase polypeptide, wherein the protein disulfide isomerase polypeptide is a PDI1 polypeptide; and

[0071] In some embodiments, the disclosure relates to a modified host cell for producing a cannabinoid or cannabinoid derivative, wherein the modified host cell comprises:

l) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise: 1) a heterologous nucleic acid comprising a nucleotide sequence encoding a chaperone or co-chaperone polypeptide, wherein the chaperone or co chaperone polypeptide is a KAR2 polypeptide; and

2) a heterologous nucleic acid comprising a nucleotide sequence encoding a protein disulfide isomerase polypeptide, wherein the protein disulfide isomerase polypeptide is a PDIl polypeptide; and

[0072] In some embodiments, the disclosure relates to a modified host cell for producing a cannabinoid or cannabinoid derivative, wherein the modified host cell comprises:

a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide;

k) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate:olivetolic acid geranyltransferase (GOT) polypeptide; and

1) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a thiol oxidase polypeptide, wherein the thiol oxidase polypeptide is an EROl polypeptide;

2) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a chaperone or co-chaperone polypeptide, wherein the chaperone or co-chaperone polypeptide is a KAR2 polypeptide;

3) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a flavin adenine dinucleotide (FAD) synthetase polypeptide, wherein the flavin adenine dinucleotide (FAD) synthetase polypeptide is a FAD1 polypeptide; and

4) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a protein disulfide isomerase polypeptide, wherein the protein disulfide isomerase polypeptide is a PDI1 polypeptide.

[0073] In some embodiments, the disclosure relates to a modified host cell for producing a cannabinoid or cannabinoid derivative, wherein the modified host cell comprises:

a) a heterologous nucleic acid comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide;

k) a heterologous nucleic acid comprising a nucleotide sequence encoding a geranyl pyrophosphate:olivetolic acid geranyltransferase (GOT) polypeptide; and l) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise: 1) a heterologous nucleic acid comprising a nucleotide sequence encoding a thiol oxidase polypeptide, wherein the thiol oxidase polypeptide is an EROl polypeptide;

2) a heterologous nucleic acid comprising a nucleotide sequence encoding a chaperone or co-chaperone polypeptide, wherein the chaperone or co chaperone polypeptide is a KAR2 polypeptide;

3) a heterologous nucleic acid comprising a nucleotide sequence encoding a flavin adenine dinucleotide (FAD) synthetase polypeptide, wherein the flavin adenine dinucleotide (FAD) synthetase polypeptide is a FADl polypeptide; and

4) a heterologous nucleic acid comprising a nucleotide sequence encoding a protein disulfide isomerase polypeptide, wherein the protein disulfide isomerase polypeptide is a PDIl polypeptide.

[0074] In some embodiments, the disclosure relates to a modified host cell for producing a cannabinoid or cannabinoid derivative, wherein the modified host cell comprises:

k) two or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate:olivetolic acid geranyltransferase (GOT) polypeptide; and

[0075] In some embodiments, the disclosure relates to a modified host cell for producing a cannabinoid or cannabinoid derivative, wherein the modified host cell comprises:

k) two heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate:olivetolic acid geranyltransferase (GOT) polypeptide; and l) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise: 1) a heterologous nucleic acid comprising a nucleotide sequence encoding a thiol oxidase polypeptide, wherein the thiol oxidase polypeptide is an EROl polypeptide;

[0076] In some embodiments, the disclosure relates to a modified host cell for producing a cannabinoid or cannabinoid derivative, wherein the modified host cell comprises:

3) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a flavin adenine dinucleotide (FAD) synthetase polypeptide, wherein the flavin adenine dinucleotide (FAD) synthetase polypeptide is a FAD1 polypeptide;

4) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a protein disulfide isomerase polypeptide, wherein the protein disulfide isomerase polypeptide is a PDI1 polypeptide;

5) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a chaperone or co-chaperone polypeptide, wherein the chaperone or co-chaperone polypeptide is a SSB1 polypeptide; and 6) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a polypeptide involved in unfolded protein response, wherein the polypeptide involved in unfolded protein response is a HACls polypeptide.

[0077] In some embodiments, the disclosure relates to a modified host cell for producing a cannabinoid or cannabinoid derivative, wherein the modified host cell comprises:

k) a heterologous nucleic acid comprising a nucleotide sequence encoding a geranyl pyrophosphate:olivetolic acid geranyltransferase (GOT) polypeptide; and 1) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise:

1) a heterologous nucleic acid comprising a nucleotide sequence encoding a thiol oxidase polypeptide, wherein the thiol oxidase polypeptide is an EROl polypeptide;

3) a heterologous nucleic acid comprising a nucleotide sequence encoding a flavin adenine dinucleotide (FAD) synthetase polypeptide, wherein the flavin adenine dinucleotide (FAD) synthetase polypeptide is a FADl polypeptide;

4) a heterologous nucleic acid comprising a nucleotide sequence encoding a protein disulfide isomerase polypeptide, wherein the protein disulfide isomerase polypeptide is a PDIl polypeptide;

5) a heterologous nucleic acid comprising a nucleotide sequence encoding a chaperone or co-chaperone polypeptide, wherein the chaperone or co chaperone polypeptide is a SSB1 polypeptide; and

6) a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in unfolded protein response, wherein the polypeptide involved in unfolded protein response is a HACls polypeptide.

[0078] In some embodiments, the disclosure relates to a modified host cell for producing a cannabinoid or cannabinoid derivative, wherein the modified host cell comprises:

a) two or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMG-CoA synthase (HMGS) polypeptide;

2) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a polypeptide involved in unfolded protein response, wherein the polypeptide involved in unfolded protein response is a HACls polypeptide.

[0079] In some embodiments, the disclosure relates to a modified host cell for producing a cannabinoid or cannabinoid derivative, wherein the modified host cell comprises:

k) a heterologous nucleic acid comprising a nucleotide sequence encoding a geranyl pyrophosphate:olivetolic acid geranyltransferase (GOT) polypeptide; and l) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise: 1) a heterologous nucleic acid comprising a nucleotide sequence encoding a chaperone or co-chaperone polypeptide, wherein the chaperone or co chaperone polypeptide is a KAR2 polypeptide; and

2) a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in unfolded protein response, wherein the polypeptide involved in unfolded protein response is a HACls polypeptide.

[0080] In some embodiments described herein, the KAR2 polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:23. In some embodiments described herein, the PDI1 polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:55. In some embodiments described herein, the EROl polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:5l. In some embodiments described herein, the IRE1 polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:67 or SEQ ID NO:578. In some embodiments described herein, the HACls polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:65. In some embodiments described herein, the FAD1 polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:37. In some embodiments described herein, the SSB1 polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: l3.

[0081] In some embodiments described herein, the modified host cell is a eukaryotic cell. In certain such embodiments, the eukaryotic cell is a yeast cell. In certain such embodiments, the yeast cell is Saccharomyces cerevisiae. In certain such embodiments, the Saccharomyces cerevisiae is a protease-deficient strain of Saccharomyces cerevisiae.

In some embodiments described herein, at least one of the one or more heterologous nucleic acids are integrated into the chromosome of the modified host cell. In some embodiments described herein, at least one of the one or more heterologous nucleic acids are maintained extrachromosomally (e.g., on a plasmid or artificial chromosome). In some embodiments described herein, at least one of the one or more heterologous nucleic acids are operably- linked to an inducible promoter. In some embodiments described herein, at least one of the one or more heterologous nucleic acids are operably-linked to a constitutive promoter. In some embodiments, at least one of the one or more heterologous nucleic acids comprises a codon-optimized nucleotide sequence.

[0082] In some embodiments, the modified host cell can produce a cannabinoid or cannabinoid derivative when cultured in a defined medium. In some embodiments, the defined medium comprises galactose, glucose, ammonium sulfate, potassium phosphate, magnesium sulfate, succinate, D-biotin, Ca-D-pantothenate, nicotinic acid, myo-inositol, thiamine hydrochloride, pyridoxal hydrochloride, p-aminobenzoic acid, EDTA, zinc sulfate heptahydrate, manganese chloride dihydrate, cobalt chloride hexahydrate, copper sulfate pentahydrate, sodium molybdate dihydrate, calcium chloride dihydrate, and iron sulfate heptahydrate.

[0083] An aspect of the disclosure relates to a method of producing a cannabinoid or a cannabinoid derivative, the method comprising culturing a modified host cell disclosed herein in a culture medium. In certain such embodiments, the method comprises recovering the produced cannabinoid or cannabinoid derivative. In some embodiments, the culture medium comprises a carboxylic acid. In certain such embodiments, the carboxylic acid is an unsubstituted or substituted C3-C 18 carboxylic acid. In certain such embodiments, the unsubstituted or substituted C3-C 18 carboxylic acid is an unsubstituted or substituted hexanoic acid. In some embodiments, the carboxylic acid is selected from the group consisting of butyric acid, valeric acid, hexanoic acid, octanoic acid, 2-methylhexanoic acid, 3-methylhexanoic acid, 4-methylhexanoic acid, 5-methylhexanoic acid, 2-hexenoic acid, 3- hexenoic acid, 4-hexenoic acid, 5-hexenoic acid, heptanoic acid, 5-chlorovaleric acid, 5- (methylsulfanyl)valeric acid, 4-pentynoic acid, trans-2-pentenoic acid, 5-hexynoic acid, trans-2-hexenoic acid, 6-heptynoic acid, trans-2-octenoic acid, nonanoic acid, trans-2- nonenoic acid, decanoic acid, undecanoic acid, dodecanoic acid, myristic acid, 4- phenylbutyric acid, 5-phenylvaleric acid, 6-phenylhexanoic acid, 7-phenylheptanoic acid, isobutyric acid, fumaric acid, itaconic acid, malic acid, succinic acid, maleic acid, malonic acid, glutaric acid, glucaric acid, oxalic acid, adipic acid, pimelic acid, suberic acid, azelaic acid, sebacic acid, dodecandioic acid, glutaconic acid, ortho-phthalic acid, isophthalic acid, terephthalic acid, citric acid, isocitric acid, aconitic acid, tricarballylic acid, trimesic acid, 5- aminovaleric acid, 5-cyanovaleric acid, 5-hydroxyvaleric acid, and 2,3-dimethylhexanoic acid, or the carboxylic acid is a carboxylic acid of Formula (II):

wherein R is a C i-C ixalkyl group substituted with R^a or R is a C2-Cisalkenyl group substituted with R^a, wherein R^a is -S-Ci-C6alkyl or -S-C2-C6alkenyl. In some embodiments, the carboxylic acid is a compound of Formula (II). In some embodiments, the culture medium comprises olivetolic acid or an olivetolic acid derivative. In some embodiments, the culture medium comprises a fermentable sugar. In some embodiments, the culture medium comprises a pretreated cellulosic feedstock. In some embodiments, the culture medium comprises a non-fermentable carbon source. In certain such embodiments, the non- fermentable carbon source comprises ethanol. In some embodiments, the culture medium is defined medium. In some embodiments, the defined medium comprises galactose, glucose, ammonium sulfate, potassium phosphate, magnesium sulfate, succinate, D-biotin, Ca-D- pantothenate, nicotinic acid, myo-inositol, thiamine hydrochloride, pyridoxal hydrochloride, p-aminobenzoic acid, EDTA, zinc sulfate heptahydrate, manganese chloride dihydrate, cobalt chloride hexahydrate, copper sulfate pentahydrate, sodium molybdate dihydrate, calcium chloride dihydrate, and iron sulfate heptahydrate.

[0084] In some embodiments described herein, the cannabinoid is cannabigerolic acid, cannabigerol, A⁹-tetrahydrocannabinolic acid, A⁹-tetrahydrocannabinol, D⁸- tetrahydrocannabinolic acid, D⁸ -tetrahydrocannabinol, cannabidiolic acid, cannabidiol, cannabichromenic acid, cannabichromene, cannabinolic acid, cannabinol, cannabidivarinic acid, cannabidivarin, tetrahydrocannabivarinic acid, tetrahydrocannabivarin,

cannabichromevarinic acid, cannabichromevarin, cannabigerovarinic acid, cannabigerovarin, cannabicyclolic acid, cannabicyclol, cannabielsoinic acid, cannabielsoin, cannabicitranic acid, or cannabicitran.

[0085] In some embodiments described herein, the cannabinoid or cannabinoid derivative is produced in an amount of more than 100 mg/L culture medium. In some embodiments described herein, the cannabinoid or cannabinoid derivative is produced in an amount of more than 50 mg/L culture medium.

[0086] One aspect of the disclosure relates to a method of making a modified host cell, the method comprising introducing into a host cell: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and b) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides. In certain such embodiments, the method comprises introducing into the host cell a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides. [0087] Another aspect of the disclosure relates to a method of making a modified host cell, the method comprising introducing into a host cell: a) one or more vectors comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and b) one or more vectors comprising one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides. In certain such embodiments, the method comprises introducing into the host cell a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides.

[0088] One aspect of the disclosure relates to compounds of Formula (I):

or pharmaceutically acceptable salts thereof, wherein,

R¹ is a Ci-Cisalkyl group substituted with R^a or R¹ is a Cri-C ixalkcnyl group substituted with R^a, wherein R^a is -S-Ci-C6alkyl or -S-Cr-Cealkeriyl, or R¹ is selected from the group consisting of:

[0089] Another aspect of the disclosure relates to compositions, including pharmaceutical compositions, comprising a compound of Formula (I), or a pharmaceutically acceptable salt thereof, and an acceptable carrier. In certain embodiments, the compositions, including pharmaceutical compositions, may comprise a compound of Formula (I), or a pharmaceutically acceptable salt thereof, and an acceptable carrier. In some embodiments, the acceptable carrier is a pharmaceutically acceptable carrier.

[0090] One aspect of the disclosure relates to methods for treating a disorder in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.

[0091] Another aspect of the disclosure relates to a methods for treating a disorder in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of a pharmaceutical composition comprising a compound of Formula (I), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0092] One aspect of the disclosure relates to uses of compounds of Formula (I), or pharmaceutically acceptable salts thereof, in the manufacture of a medicament for treating a disorder in a subject in need thereof.

[0093] Another aspect of the disclosure relates to uses of pharmaceutical

compositions comprising a compound of Formula (I), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier in the manufacture of a medicament for treating a disorder in a subject in need thereof.

[0094] One aspect of the disclosure relates to uses of compounds of Formula (I), or pharmaceutically acceptable salts thereof, for treating a disorder in a subject in need thereof.

[0095] Another aspect of the disclosure relates to uses of pharmaceutical

compositions comprising a compound of Formula (I), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier for treating a disorder in a subject in need thereof.

[0096] One aspect of the disclosure relates to compounds of Formula (I), or pharmaceutically acceptable salts thereof, for use in methods of treating a disorder in a subject in need thereof.

[0097] Another aspect of the disclosure relates to a pharmaceutical composition comprising a compound of Formula (I), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier for use in a method of treating a disorder in a subject in need thereof.

[0098] In some embodiments of the methods, uses, compounds for use,

pharmaceutical compositions for use, and medicaments described herein, the disorder is mediated by cannabinoid receptor type 1 or cannabinoid receptor type 2. In some embodiments of the methods, uses, compounds for use, pharmaceutical compositions for use, and medicaments described herein, the disorder is chronic pain, multiple sclerosis, cancer-associated nausea and vomiting, weight loss, appetite loss, spasticity, or seizures. BRIEF DESCRIPTION OF THE DRAWINGS

[0099] FIGS. 1A, IB, and 1C depict expression constructs used in the production of the S29 strain. The expression constructs depicted in FIGS. 1 A, 1B, and 1C were also used in the production of the following strains: S34, S61, S76, S122, S 131 , S149, S 171, S 181 , S184, S185, SI 94, S196, S198, S200, S201, S206, S208, S209, S210, S211, S213, S220,

S222, S223, S224, S228, S230, S233, S241, S245, S253, S254, S255, S258, S259, S268,

S269, S270, S271, S272, S276, S277, S305, S306, S307, S308, S311, S312, S313, S314,

S315, S316, S317, S318, S319, S320, S321, S322, S323, S324, S325, S326, S327, S328,

S336, S337, S347, S348, S349, S350, S351, S353, S354, S355, S356, S357, S359, S360,

S361, S362, S369, S487, S494, S497, S499, S510, S542, S579, S699-S791, S935, S938, S940-S946, S951, S1000-S1059, S1072-1079, S1081, SI 100-S1120, and S1205-S1208. Throughout the figures, in addition to the specified coding sequences from Table 1, construct maps depict regulatory, non-coding and genomic cassette sequences described in Table 10. Construct maps also depict genes denoted with a preceding“m” (e.g., mERG13), which specify open reading frames from Table 1 with 200-250 base pairs (bp) of downstream regulatory (terminator) sequence. Arrows in construct maps indicate the directionality of certain DNA parts. The“!” preceding a part name is an output of the DNA design software used, is redundant with the arrow directionality, and can be ignored.

[00100] FIG. 2 depicts expression constructs used in the production of the S131 strain.

[00101] FIG. 3 depicts an expression construct used in the production of the S181 strain. The expression construct depicted in FIG. 3 was also used in the production of following strains: S220, S222, S233, S241, S245, S259, S268, S269, S270, S271, S272,

S327, S328, S347, S348, S349, S350, S351, S369, S487, S579, S699-S791, S935, S938, S940-S946, S951, S1000-S1059, S1072-1079, S1081, and S1205-S1208.

[00102] FIG. 4 depicts an expression construct used in the production of the S220 strain. The expression construct depicted in FIG. 4 was also used in the production of following strains: S233, S241, S245, S259, S268, S269, S270, S271, S272, S327, S328,

S347, S348, S349, S350, S351, S369, S487, S579, S699-S791, S935, S938, S940-S946,

S951, S1000-S1059, S1072-1079, S1081, and S1205-S1208.

[00103] FIGS. 5A and 5B depict expression constructs used in the production of the S233 strain. The expression constructs depicted in FIGS. 5A and 5B were also used in the production of the S259 strain.

[00104] FIG. 6 depicts expression constructs used in the production of the S241 strain. The expression constructs depicted in FIG. 6 were also used in the production of following strains: S268, S269, S270, S271, S272, S327, S328, S347, S348, S349, S350, S351, S369, S487, S579, S699-S791, S935, S938, S940-S946, S951, S1000-S1059, S1072-1079, S1081, and S1205-S1208.

[00105] FIGS. 7A and 7B depict expression constructs used in the production of the S259 strain.

[00106] FIG. 8 depicts expression constructs used in the production of the S268 strain.

[00107] FIG. 9 depicts expression constructs used in the production of the S271 strain.

[00108] FIG. 10 depicts expression constructs used in the production of the S272 strain.

[00109] FIG. 11 depicts a landing pad construct used in the production of the S61 strain. The construct depicted in FIG. 11 was also used in the production of the following strains: S122, S171, S181, S184, S185, S194, S196, S198, S200, S201, S208, S209, S210, S211, S213, S220, S222, S223, S224, S230, S233, S241, S245, S253, S258, S259, S268,

S269, S270, S271, S272, S276, S277, S305, S306, S307, S308, S327, S328, S336, S337,

S951, S1000-S1059, S1072-1079, S1081, and S1205-S1208.

[00110] FIG. 12 depicts expression constructs used in the production of the S122 strain. The expression constructs depicted in FIG. 12 were also used in the production of the following strains: S171, S181, S184, S185, S194, S196, S198, S200, S201, S208, S209, S210, S211, S213, S220, S222, S223, S224, S230, S233, S241, S245, S253, S258, S259,

S268, S269, S270, S271, S272, S276, S277, S305, S306, S307, S308, S327, S328, S336,

S337, S347, S348, S349, S350, S351, S369, S487, S579, S699-S791, S935, S938, S940- S946, S951, S1000-S1059, S1072-1079, S1081, and S1205-S1208.

[00111] FIG. 13 depicts an expression construct used in the production of the S171 strain. The expression construct depicted in FIG. 13 was also used in the production of the following strains: S181, S194, S196, S198, S200, S201, S208, S209, S210, S211, S213, S220, S222, S223, S224, S230, S233, S241, S245, S253, S258, S259, S268, S269, S270,

S271, S272, S276, S277, S305, S306, S307, S308, S327, S328, S336, S337, S347, S348,

S349, S350, S351, S369, S487, S579, S699-S791, S935, S938, S940-S946, S951, S1000- S1059, S1072-1079, S1081, and S1205-S1208.

[00112] FIG. 14 depicts expression constructs used in the production of the S201 strain. The expression constructs depicted in FIG. 14 were also used in the production of the following strains: S230, S253, S258, S276, S277, S305, S306, S307, S308, S336, and S337. [00113] FIG. 15 depicts expression constructs used in the production of the S213 strain.

[00114] FIGS. 16A and 16B depict expression constructs used in the production of the S230 strain. The expression constructs depicted in FIGS. 16A and 16B were also used in the production of the S253 strain.

[00115] FIGS. 17A and 17B depict expression constructs used in the production of the

5253 strain.

[00116] FIGS. 18A and 18B depict expression constructs used in the production of the S206 strain. The expression constructs depicted in FIGS. 18A and 18B were also used in the production of the following strains: S228, S254, S255, S318, S319, S320, S321, S322, S323, S324, S326, S359, S360, S361, S362, S494, S497, S499, S510, S542, and SI 100-SI 120.

[00117] FIGS. 19A and 19B depict expression constructs used in the production of the

5254 strain.

[00118] FIG. 20 depicts an expression construct used in the production of the S184 strain.

[00119] FIG. 21 depicts an expression construct used in the production of the S185 strain.

[00120] FIGS. 22A and 22B depict expression constructs used in the production of the S194 strain.

[00121] FIG. 23 depicts expression constructs used in the production of the S196 strain.

[00122] FIG. 24 depicts expression constructs used in the production of the SI 98 strain.

[00123] FIG. 25 depicts expression constructs used in the production of the S200 strain.

[00124] FIG. 26 depicts expression constructs used in the production of the S208 strain.

[00125] FIG. 27 depicts expression constructs used in the production of the S209 strain.

[00126] FIG. 28 depicts expression constructs used in the production of the S210 strain.

[00127] FIG. 29 depicts expression constructs used in the production of the S211 strain. [00128] FIG. 30 depicts expression constructs used in the production of the S223 strain.

[00129] FIG. 31 depicts expression constructs used in the production of the S224 strain.

[00130] FIG. 32 depicts expression constructs used in the production of the S222 strain.

[00131] FIG. 33 depicts expression constructs used in the production of the S245 strain.

[00132] FIG. 34 depicts expression constructs used in the production of the S258 strain.

[00133] FIG. 35 depicts expression constructs used in the production of the S269 strain.

[00134] FIG. 36 depicts expression constructs used in the production of the S270 strain. The expression constructs depicted in FIG. 36 were also used in the production of the following strains: S327, S328, S347, S348, S349, S350, S351, S369, S487, S579, S699- S791, S935, S938, S940-S946, S951, S1000-S1059, S1072-1079, S1081, and S1205-S1208.

[00135] FIG. 37 depicts expression constructs used in the production of the S305 strain.

[00136] FIG. 38 depicts expression constructs used in the production of the S306 strain.

[00137] FIG. 39 depicts expression constructs used in the production of the S307 strain.

[00138] FIG. 40 depicts expression constructs used in the production of the S308 strain.

[00139] FIG. 41 depicts expression constructs used in the production of the S228 strain. The expression constructs depicted in FIG. 41 were also used in the production of the following strains: S318, S319, S320, S321, S322, S323, S324, S326, S359, S360, S361, and S362.

[00140] FIG. 42 depicts an expression construct used in the production of the S311 strain.

[00141] FIG. 43 depicts an expression construct used in the production of the S312 strain.

[00142] FIG. 44 depicts an expression construct used in the production of the S313 strain. [00143] FIG. 45 depicts an expression construct used in the production of the S314 strain.

[00144] FIG. 46 depicts an expression construct used in the production of the S315 strain.

[00145] FIG. 47 depicts an expression construct used in the production of the S316 strain.

[00146] FIG. 48 depicts an expression construct used in the production of the S317 strain.

[00147] FIG. 49 depicts an expression construct used in the production of the S318 strain.

[00148] FIG. 50 depicts an expression construct used in the production of the S319 strain.

[00149] FIG. 51 depicts an expression construct used in the production of the S320 strain.

[00150] FIG. 52 depicts an expression construct used in the production of the S321 strain.

[00151] FIG. 53 depicts an expression construct used in the production of the S322 strain.

[00152] FIG. 54 depicts an expression construct used in the production of the S323 strain.

[00153] FIG. 55 depicts an expression construct used in the production of the S324 strain.

[00154] FIG. 56 depicts expression constructs used in the production of the S325 strain.

[00155] FIG. 57 depicts expression constructs used in the production of the S326 strain.

[00156] FIG. 58 depicts expression constructs used in the production of the S327 strain.

[00157] FIG. 59 depicts expression constructs used in the production of the S328 strain.

[00158] FIG. 60 depicts expression constructs used in the production of the S336 strain.

[00159] FIG. 61 depicts expression constructs used in the production of the S337 strain. [00160] FIG. 62 depicts spectral data of cannabidivarinic acid produced by the S253 strain after butanoic acid feeding.

[00161] FIG. 63 depicts spectral data of 6-butyl-2,4-dihydroxy-3- [(lR,6R)-3-methyl-6-(prop-l-en-2-yl)cyclohex-2-en-l-yl]benzoic acid produced by the S253 strain after pentanoic acid feeding.

[00162] FIG. 64 depicts spectral data of 6-hexyl-2,4-dihydroxy-3-[(lR,6R)-3-methyl- 6-(prop-l-en-2-yl)cyclohex-2-en-l-yl]benzoic acid produced by the S253 strain after heptanoic acid feeding.

[00163] FIG. 65 depicts expression constructs used in the production of the S276 strain.

[00164] FIG. 66 depicts expression constructs used in the production of the S277 strain. Genes shown in FIGS. 65 and 66 are genomic expression cassettes, including 500 bp of upstream (promoter) sequence, the open reading frame, and 250 bp of downstream (terminator) sequence.

[00165] FIG. 67 depicts expression constructs used in the production of the S369 strain. The expression constructs depicted in FIG. 67 were also used in the production of the following strains: S347, S348, S349, S350, and S351.

[00166] FIG. 68 depicts an expression construct used in the production of the S347 strain.

[00167] FIG. 69 depicts an expression construct used in the production of the S348 strain.

[00168] FIG. 70 depicts an expression construct used in the production of the S349 strain.

[00169] FIG. 71 depicts an expression construct used in the production of the S350 strain.

[00170] FIG. 72 depicts an expression construct used in the production of the S351 strain.

[00171] FIG. 73 depicts an expression construct used in the production of the S353 strain.

[00172] FIG. 74 depicts an expression construct used in the production of the S354 strain.

[00173] FIG. 75 depicts an expression construct used in the production of the S355 strain. [00174] FIG. 76 depicts an expression construct used in the production of the S356 strain.

[00175] FIG. 77 depicts an expression construct used in the production of the S357 strain.

[00176] FIG. 78 depicts microscopy images of the secretory engineering-naive strain S29 expressing CBDASco5-GFP (left panel = S325) and the secretory engineered strain S228 expressing CBDASco5-GFP (right panel = S326). Green fluorescent protein (GFP) fluorescence appears in white against gray/black background. Small amounts of GFP in sharp foci are visible in the left panel, compared to diffuse and distributed GFP in the right panel.

[00177] FIG. 79 depicts a graphical representation of Table 5 data. CBDA titer (y- axis) versus GFP polypeptide expression level (x-axis) of strains expressing various codon optimizations of CBDAS fused via T2A linker to GFP. Data in the left panel are from strains S311-S317; right panel from S318-S324.

[00178] FIG. 80 depicts an expression construct used in the production of the S359 strain.

[00179] FIG. 81 depicts an expression construct used in the production of the S360 strain.

[00180] FIG. 82 depicts an expression construct used in the production of the S361 strain.

[00181] FIG. 83 depicts expression constructs used in the production of the S362 strain.

[00182] FIG. 84 depicts expression constructs used in the production of the S34 strain. The expression constructs depicted in FIG. 84 were also used in the production of the following strains: S76 and S149.

[00183] FIG. 85 depicts an expression construct used in the production of the S76 strain. The expression construct depicted in FIG. 85 was also used in the production of the SI 49 strain.

[00184] FIG. 86 depicts an expression construct used in the production of the S149 strain.

[00185] FIG. 87 depicts an expression construct used in the production of the S255 strain. The expression construct depicted in FIG. 87 was also used in the production of the following strains: S494, S497, S499, and S542. [00186] FIG. 88 depicts expression constructs used in the production of the S494 strain.

[00187] FIG. 89 depicts expression constructs used in the production of the S497 strain.

[00188] FIG. 90 depicts expression constructs used in the production of the S499 strain.

[00189] FIG. 91 depicts expression constructs used in the production of the S542 strain.

[00190] FIG. 92 depicts expression constructs used in the production of the S487 strain. The expression constructs depicted in FIG. 92 were also used in the production of the following strains: S579, S699-S791, S935, S938, S940-S946, S951, S1000-S1059, S1072- 1079, S 1081 , and S1205-S1208.

[00191] FIG. 93 depicts an expression construct used in the production of the S579 and S1100 strains.

[00192] FIG. 94 depicts an expression construct used in the production of the S699- S791, S935, S938, S940-S946, Sl 101-1120, and S1205-S1208 strains.

[00193] FIG. 95 depicts an expression construct used in the production of the SI 042 strain.

[00194] FIG. 96 depicts an expression construct used in the production of the following strains: S951, S1000-S1041, S1043-S1059, S1072-1079, and S1081.

[00195] FIG. 97 depicts an expression construct used in the production of the S510 strain. The expression construct depicted in FIG. 97 was also used in the production of the following strains: S1100-S1120.

DETAILED DESCRIPTION

[00196] Synthetic biology allows for the engineering of industrial host organisms— e.g., microbes— to convert simple sugar feedstocks into medicines. This approach includes identifying genes that produce the target molecules and optimizing their activities in the industrial host. Microbial production can be significantly cost-advantaged over agriculture and chemical synthesis, less variable, and allow tailoring of the target molecule. However, reconstituting or creating a pathway to produce a target molecule in an industrial host organism can require significant engineering of both the pathway genes and the host. The present disclosure provides modified host cells comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide. The modified host cells of the disclosure may be useful for producing cannabinoids or cannabinoid derivatives (e g., non-naturally occurring cannabinoids) and/or for expressing a cannabinoid synthase polypeptide. The present disclosure also provides methods, nucleic acids, and modified host cells for producing cannabinoids or cannabinoid derivatives.

Additionally, the present disclosure provides methods, nucleic acids, and modified host cells for expressing a cannabinoid synthase polypeptide. Further, the present disclosure provides novel cannabinoids and cannabinoid derivatives, compositions, including pharmaceutical compositions, as well as methods, uses, compounds for use, medicaments, and compositions (including pharmaceutical compositions) for use comprising said cannabinoid and cannabinoid derivatives.

[00197] Cannabinoid synthase polypeptides, such as tetrahydrocannabinolic acid synthase, cannabichromenic acid synthase, or cannabidiolic acid synthase polypeptides, play an important role in the biosynthesis of cannabinoids. However, reconstituting their activity in a modified host cell has proven challenging, hampering progress in the production of cannabinoids or cannabinoid derivatives.

[00198] Herein, nucleic acids comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide are disclosed. Surprisingly, expression of the cannabinoid synthase polypeptides in a host cell modified with one or more nucleic acids comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide did not significantly decrease the growth or viability of the modified host cell compared to an unmodified host cell. Commonly, expression of cannabinoid synthase polypeptides by host cells modified to express these polypeptides is toxic to the host cell, reducing viability and resulting in cell death. Aggregation of the cannabinoid synthase polypeptides within the modified host cell is often observed. Nucleic acids disclosed herein comprising a codon-optimized nucleotide sequence encoding said cannabinoid synthase polypeptides are useful in the methods and modified host cells of the disclosure for producing cannabinoids or cannabinoid derivatives and for expressing cannabinoid synthase polypeptides.

[00199] The disclosure also provides for modification of the secretory pathway of a host cell modified with one or more nucleic acids (e.g., heterologous nucleic acids) comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide. In some embodiments, the nucleotide sequence encoding a cannabinoid synthase polypeptide is codon-optimized. Modification of the secretory pathway in the host cell may improve expression and solubilization of cannabinoid synthase polypeptides, as these polypeptides are processed through the secretory pathway. Reconstituting the activity of polypeptides processed through the secretory pathway, such as cannabinoid synthase polypeptides, in a modified host cell, such as a modified yeast cell, can be challenging and unreliable. Often the expressed polypeptides are misfolded or mislocalized, resulting in low expression, expressed polypeptides lacking activity, polypeptide aggregation, reduced host cell viability, and/or cell death. Additionally, a backlog of misfolded or mislocalized expressed polypeptides can induce metabolic stress within the modified host cell, harming the modified host cell. The expressed polypeptides may lack necessary posttranslational modifications for folding and activity, such as disulfide bonds, glycosylation and trimming, and cofactors, affording inactive polypeptides or polypeptides with reduced enzymatic activity. Moreover, the addition of a signal sequence polypeptide to the cannabinoid synthase polypeptides, such as a secretory signal sequence polypeptide, may facilitate proper transport of cannabinoid synthase polypeptides through the secretory pathway, also possibly improving host cell viability and expression, solubilization, folding, and enzymatic activity of the cannabinoid synthase polypeptides.

[00200] The methods of the disclosure may include using microorganisms engineered (e.g., modified host cells) to produce naturally-occurring and non-naturally occurring cannabinoids. Naturally-occurring cannabinoids and non-naturally occurring cannabinoids (e g., cannabinoid derivatives) are challenging to produce using chemical synthesis due to their complex structures. The methods of the disclosure enable the construction of metabolic pathways inside living cells to produce bespoke cannabinoids or cannabinoid derivatives from simple precursors such as sugars and carboxylic acids. One or more nucleic acids (e.g., heterologous nucleic acids) disclosed herein encoding one or more polypeptides disclosed herein can be introduced into host microorganisms allowing for the stepwise conversion of inexpensive feedstocks, e.g., sugar, into final products: cannabinoids or cannabinoid derivatives. These products can be specified by the choice and construction of expression constructs or vectors comprising one or more nucleic acids (e.g., heterologous nucleic acids) disclosed herein, allowing for the efficient bioproduction of chosen cannabinoids, such as THC or CBD and less common cannabinoid species found at low levels in Cannabis ; or cannabinoid derivatives. Bioproduction also enables synthesis of cannabinoids or cannabinoid derivatives with defined stereochemistries, which is challenging to do using chemical synthesis. To produce cannabinoids or cannabinoid derivatives and create biosynthetic pathways within modified host cells, modified host cells comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide may express or overexpress combinations of heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cannabinoid or cannabinoid precursor (e g., geranylpyrophosphate (GPP), prenyl phosphates, olivetolic acid, or hexanoyl-CoA) biosynthesis. In some embodiments, the nucleotide sequences encoding the one or more polypeptides involved in cannabinoid or cannabinoid precursor biosynthesis are codon-optimized.

[00201] The modified host cell of the disclosure may be a modified yeast cell. Yeast cells may be reasonable host cells for protein expression as well as for cannabinoid and cannabinoid derivative production because they are cultured using known conditions, grow rapidly, and are generally regarded as safe. Yeast cells contain the secretory pathway common to all eukaryotes. As disclosed herein, manipulation of that secretory pathway in yeast host cells modified with one or more nucleic acids (e.g., heterologous nucleic acids) comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide may improve expression, folding, and enzymatic activity of the cannabinoid synthase polypeptide as well as viability of the modified yeast host cell, such as modified Saccharomyces cerevisiae. Further, addition of a signal sequence polypeptide to the cannabinoid synthase polypeptides, such as a secretory signal sequence polypeptide, and/or use of codon- optimized nucleotide sequences encoding cannabinoid synthase polypeptides, may improve expression and activity of the cannabinoid synthase polypeptide and viability of modified yeast host cells, such as modified Saccharomyces cerevisiae.

[00202] Besides allowing for the production of desired cannabinoids or cannabinoid derivatives, the present disclosure provides a more reliable and economical process than agriculture-based production. Microbial fermentations can be completed in days versus the months necessary for an agricultural crop, are not affected by climate variation or soil contamination (e.g., by heavy metals), and can produce pure products at high titer.

[00203] The present disclosure also provides a platform for the economical production of high-value cannabinoids including THC and CBD, as well as derivatives thereof. It also provides for the production of different cannabinoids or cannabinoid derivatives for which no viable method of production exists. Using the methods and modified host cells disclosed herein, cannabinoids and cannabinoid derivatives may be produced in an amount of over 100 mg per liter of culture medium, over 1 g per liter of culture medium, over 10 g per liter of culture medium, or over 100 g per liter of culture medium. [00204] Additionally, the disclosure provides methods, modified host cells, and nucleic acids to produce cannabinoids or cannabinoid derivatives in vivo or in vitro from simple precursors. Nucleic acids (e.g., heterologous nucleic acids) disclosed herein can be introduced into microorganisms (e.g., modified host cells), resulting in expression or overexpression of one or more polypeptides, which can then be utilized in vitro or in vivo for the production of cannabinoids or cannabinoid derivatives. In some embodiments, the in vitro methods are cell-free.

Cannabinoid Biosynthesis

[00205] In addition to one or more nucleic acids (e.g., heterologous nucleic acids) encoding a cannabinoid synthase polypeptide, one or more nucleic acids (e.g., heterologous nucleic acids) encoding one or more polypeptides having at least one activity of a polypeptide present in the cannabinoid or cannabinoid precursor (e.g., geranylpyrophosphate (GPP), prenyl phosphates, olivetolic acid, or hexanoyl-CoA) biosynthetic pathway may be useful in the methods and modified host cells for the synthesis of cannabinoids or cannabinoid derivatives. In some embodiments, the nucleotide sequences encoding the one or more polypeptides involved in cannabinoid or cannabinoid precursor biosynthesis are codon-optimized.

[00206] In Cannabis, cannabinoids are produced from the common metabolite precursors geranylpyrophosphate (GPP) and hexanoyl-CoA by the action of three polypeptides. Hexanoyl-CoA and malonyl-CoA are combined to afford a 12-carbon tetraketide intermediate by a tetraketide synthase (TKS) polypeptide. This tetraketide intermediate is then cyclized by an olivetolic acid cyclase (OAC) polypeptide to produce olivetolic acid. Olivetolic acid is then prenylated with the common isoprenoid precursor GPP by a geranyl pyrophosphate: olivetolic acid geranyltransferase (GOT) polypeptide (e.g., a CsPT4 polypeptide) to produce cannabigerolic acid (CBGA), the cannabinoid also known as the“mother cannabinoid.” Different cannabinoid synthase polypeptides then convert CBGA into other cannabinoids, e.g., a tetrahydrocannabinolic acid (THCA) synthase polypeptide produces THCA, a CBDA synthase polypeptide produces CBDA, etc. In the presence of heat or light, the acidic cannabinoids can undergo decarboxylation, e.g., THCA producing THC or CBDA producing CBD.

[00207] GPP and hexanoyl-CoA can be generated through several pathways. One or more nucleic acids (e.g., heterologous nucleic acids) encoding one or more polypeptides having at least one activity of a polypeptide present in these pathways can be useful in the methods and modified host cells for the synthesis of cannabinoids or cannabinoid derivatives.

[00208] Polypeptides that generate GPP or are part of a biosynthetic pathway that generates GPP may be one or more polypeptides having at least one activity of a polypeptide present in the mevalonate (MEV) pathway (e.g., one or more MEV pathway polypeptides). The term“mevalonate pathway” or“MEV pathway,” as used herein, may refer to the biosynthetic pathway that converts acetyl-CoA to isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP). The mevalonate pathway comprises polypeptides that catalyze the following steps: (a) condensing two molecules of acetyl-CoAto generate acetoacetyl-CoA (e.g., by action of an acetoacetyl-CoAthiolase polypeptide); (b) condensing acetoacetyl-CoA with acetyl-CoA to form hydroxymethylglutaryl-CoA (HMG-CoA) (e.g., by action of a HMG-CoA synthase (HMGS) polypeptide); (c) converting HMG-CoA to mevalonate (e.g., by action of a HMG-CoA reductase (HMGR) polypeptide); (d) phosphorylating mevalonate to mevalonate 5-phosphate (e.g., by action of a mevalonate kinase (MK) polypeptide); (e) converting mevalonate 5-phosphate to mevalonate 5- pyrophosphate (e.g., by action of a phosphomevalonate kinase (PMK) polypeptide); (f) converting mevalonate 5-pyrophosphate to isopentenyl pyrophosphate (e.g., by action of a mevalonate pyrophosphate decarboxylase (MVD1) polypeptide); and (g) converting isopentenyl pyrophosphate (IPP) to dimethylallyl pyrophosphate (DMAPP) (e.g., by action of an isopentenyl pyrophosphate isomerase (IDI1) polypeptide) A geranyl pyrophosphate synthetase (GPPS) polypeptide then acts on IPP and/or DMAPP to generate GPP.

[00209] Polypeptides that generate hexanoyl-CoA may include polypeptides that generate acyl-CoA compounds or acyl-CoA compound derivatives (e.g., a hexanoyl-CoA synthase (HCS) polypeptide, an acyl-activating enzyme polypeptide, a fatty acyl-CoA synthetase polypeptide, or a fatty acyl-CoA ligase polypeptide). Hexanoyl CoA derivatives, acyl-CoA compounds, or acyl-CoA compound derivatives may also be formed via such polypeptides.

[00210] GPP and hexanoyl-CoA may also be generated through pathways comprising polypeptides that condense two molecules of acetyl-CoA to generate acetoacetyl-CoA and pyruvate decarboxylase polypeptides that generate acetyl-CoA from pyruvate via acetaldehyde. Hexanoyl CoA derivatives, acyl-CoA compounds, or acyl-CoA compound derivatives may also be formed via such pathways.

Secretory Pathway

[00211] Cannabinoids are produced via polypeptides, e.g., cannabinoid synthase polypeptides, processed through the secretory pathway— a diverse set of transport and modification routes for cargo polypeptides that can be different for different polypeptides and organisms. Secretory processing typically includes initial transport into the endoplasmic reticulum (ER), where a cargo polypeptide can undergo folding and refolding, disulfide- bond formation and isomerization, proline isomerization, flavin cofactor binding, various forms of glycosylation and trimming, and other modifications. From the ER, secretory cargo polypeptides can be targeted to various organelles or extracellular localizations where they can undergo additional processing. For many polypeptides, processing along the secretory pathway is crucial for their maturation. Functionally reconstituting the activity of cannabinoid synthase polypeptides in a heterologous host cell can require engineering the host cell’s secretory pathway and tuning the expression of the target polypeptide(s).

[00212] In eukaryotic cells, including yeast, polypeptides destined for the ER, the Golgi, the vacuole and the extracellular space are processed and sorted through the secretory pathway (reviewed extensively in Barlowe and Miller, 2013; Delic et al. 2013). The process begins in the cytoplasm where translation of secretory polypeptide mRNAs is initiated. The polypeptide is then translocated to the ER, either co-translationally (via interaction between the ribosome-bound native polypeptide and the signal recognition particle (SRP) complex), or post-translationally (via the SEC63 complex with recruitment by cytosolic chaperones). In both cases, the polypeptide enters the ER through the SEC61 pore complex. As the polypeptide enters the ER, it interacts first with the chaperone KAR2, a luminal HSP70 ATPase that assists with the translocation and folding of the polypeptide. Signal sequence processing by the Signal Peptidase Complex (SPC) and the attachment of asparagine-linked (N-linked) glycosylations by the oligosaccharyltransferase (OST) enzymes also occur during translocation.

[00213] Once inside the ER, further maturation of the polypeptide occurs. Some polypeptides undergo O-linked glycosylation on serine and threonine residues and some are modified on their carboxy-terminus by attachment of a lipid-anchored

glycosylphosphatidylinositol (GPI) moiety. Proper disulfide bond formation is catalyzed by disulfide isomerases such as PDI1, which require oxidizing equivalents from polypeptides such as EROl or ERV2. KAR2 continues to play a chaperone role in maturation and peptidyl-prolyl isomerases such as CPR5 are another class of chaperone that assists with polypeptide folding at this stage. Processing of the initial l4-residue N-linked glycosylations by glucosidases such as ROT2 plays an important role in quality control, and the polypeptide may enter a calnexin (CNEl)-dependent folding cycle, where improperly folded

polypeptides are modified such that they remain bound to CNE1, while properly folded substrates acquire a specific glycosylation pattern that allows exit from the cycle. In the case of terminally misfolded polypeptides, the glycosylations are trimmed by the mannosidases MNS1 and HTM1, allowing interaction with the ER lectin YOS9 and targeting for ER- associated degradation (ERAD). ERAD comprises a series of pathways that remove misfolded polypeptides for ubquitination and degradation in the proteasome.

[00214] These complex ER functions and capabilities are regulated by a

transcriptional program known as the Unfolded Protein Response (UPR). Accumulation of unfolded polypeptides is initially sensed by association between the lumenal sensor polypeptides IRE1 and KAR2, formation of oligomers and activation of the kinase and ribonuclease activity of IRE1, which splices HAC1 mRNA into the mature form. The transcription factor HAC1 is a strong activator of genes containing UPR elements in their regulatory regions, and in response to ER stress it upregulates functions that save as well as destroy misfolded polypeptides.

[00215] Following maturation in the ER, secretory forward traffic involves assembly (via interaction with SEC24) into COPII-protein complex coated transport vesicles that cross the cytoplasm to fuse with the Golgi via the activity of SNARE complexes. Inside the Golgi, further processing of the glycosylation chain and/or proteolytic processing may occur.

Polypeptides bearing an ER retention signal (HDEL or KDEL signal) may be returned to the ER in COPI-coated vesicles, a process known as retrograde transport. Polypeptides destined for the vacuole may be routed to the endosome by the sorting receptor PEP1. Finally, polypeptides destined for the extracellular space move to the trans-face of the Golgi and are tethered to the plasma membrane by the exocyst, where SNARE proteins are once again involved in membrane fusion and release of the secreted polypeptide.

[00216] In the Cannabis plant, cannabinoid synthase polypeptides are processed through the secretory pathway and secreted into the glandular trichome. These polypeptides contain a disulfide bridge and have multiple glycosylation sites, features that pose a challenge for expression in yeast. As disclosed herein, engineering of the yeast secretory pathway may achieve substantial cannabinoid synthase polypeptide function, expression, and solubilization in yeast.

General Information

[00217] In certain aspects, the practice of the present disclosure will employ, unless otherwise indicated, conventional techniques of molecular biology (including recombinant techniques), microbiology, cell biology, biochemistry, and immunology, which are within the skill of the art. Such techniques are explained fully in the literature:“ Molecular

Cloning: A Laboratory Manual," second edition (Sambrook et al., 1989);“ Oligonucleotide Synthesis’’ (M. J. Gait, ed., 1984);“ Animal Cell Culture” (R. I. Freshney, ed., 1987);

“ Methods in Enzymology” (Academic Press, Inc.);“Current Protocols in Molecular

Biology” (F. M. Ausubel et al., eds., 1987, and periodic updates);“PCR: The Polymerase Chain Reaction (Mullis et al., eds., 1994). Singleton et al., Dictionary of Microbiology and Molecular Biology 2nd ed., J. Wiley & Sons (New York, N.Y. 1994), and March, Advanced Organic Chemistry Reactions, Mechanisms and Structure 4th ed., John Wiley & Sons (New York, N.Y. 1992), provide one skilled in the art with a general guide to many of the terms used in the present application.

[00218] “Cannabinoid” or“cannabinoid compound” as used herein may refer to a member of a class of unique meroterpenoids found until now only in Cannabis sativa.

Cannabinoids may include, but are not limited to, cannabichromene (CBC) type (e.g., cannabichromenic acid), cannabigerol (CBG) type (e.g., cannabigerolic acid), cannabidiol (CBD) type (e.g., cannabidiolic acid), A⁹-trans-tetrahydrocannabinol (D⁹ -THC) type (e.g., A⁹-tetrahydrocannabinolic acid), A⁸-trans-tetrahydrocannabinol (D⁸ -THC) type,

cannabicyclol (CBL) type, cannabielsoin (CBE) type, cannabinol (CBN) type, cannabinodiol (CBND) type, cannabitriol (CBT) type, cannabigerolic acid (CBGA), cannabigerolic acid monomethylether (CBGAM), cannabigerol (CBG), cannabigerol monomethylether

(CBGM), cannabigerovarinic acid (CBGYA), cannabigerovarin (CBGV), cannabichromenic acid (CBCA), cannabichromene (CBC), cannabichromevarinic acid (CBCVA),

cannabichromevarin (CBCV), cannabidiolic acid (CBDA), cannabidiol (CBD), cannabidiol monomethylether (CBDM), cannabidiol-Cri (CBD-C4), cannabidivarinic acid (CBDVA), cannabidivarin (CBDV), cannabidiorcol (CBD-Ci), D⁹ -tetrahydrocannabinolic acid A (THCA-A), D⁹ -tetrahydrocannabinolic acid B (THCA-B), D⁹ -tetrahydrocannabinol (THC), D⁹ -tetrahydrocannabinolic acid-Cri (THCA-C4), D⁹ -tetrahydrocannabinol-C₄ (THC-C4), D⁹ -tetrahydrocannabivarinic acid (THCVA), D⁹ -tetrahydrocannabivarin (THCV), D⁹ - tetrahydrocannabiorcolic acid (THCA-Ci), D⁹ -tetrahydrocannabiorcol (THC-Ci), D⁷ -cis- iso-tetrahydrocannabivarin, D⁸ -tetrahydrocannabinolic acid (A⁸ -THCA), A⁸ - tetrahydrocannabinol (A⁸ -THC), cannabicyclolic acid (CBLA), cannabicyclol (CBL), cannabicyclovarin (CBLV), cannabielsoic acid A (CBEA-A), cannabielsoic acid B (CBEA- B), cannabielsoin (CBE), cannabielsoinic acid, cannabicitranic acid, cannabinolic acid (CBNA), cannabinol (CBN), cannabinol methylether (CBNM), cannabinol-C4, (CBN-C₄), cannabivarin (CBV), cannabinol-C2 (CNB-C2), cannabiorcol (CBN-Ci), cannabinodiol (CBND), cannabinodivarin (CBVD), cannabitriol (CBT), 10-ethyoxy-9-hydroxy-delta-6a- tetrahydrocannabinol, 8,9-dihydroxyl-delta-6a-tetrahydrocannabinol, cannabitriolvarin (CBTVE), dehydrocannabifuran (DCBF), cannabifuran (CBF), cannabichromanon (CBCN), cannabicitran (CBT), l0-oxo-delta-6a-tetrahydrocannabinol (OTHC), delta-9-cis- tetrahydrocannabinol (cis-THC), 3,4,5,6-tetrahydro-7-hydroxy-alpha-alpha-2-trimethyl-9-n- propyl-2,6-methano-2H-l-benzoxocin-5-methanol (OH-iso-HHCV), cannabiripsol (CBR), and trihydroxy-delta-9-tetrahydrocannabinol (triOH-THC).

[00219] An acyl-CoA compound as detailed herein may include compounds with the

O

following structure

wherein R may be an unsubstituted fatty acid side chain or a fatty acid side chain substituted with or comprising one or more functional and/or reactive groups as disclosed herein (i.e., an acyl-CoA compound derivative).

[00220] As used herein, a hexanoyl CoA derivative, an acyl-CoA compound derivative, a cannabinoid derivative, or an olivetolic acid derivative may refer to hexanoyl CoA, an acyl-CoA compound, a cannabinoid, or olivetolic acid substituted with or comprising one or more functional and/or reactive groups. Functional groups may include, but are not limited to, azido, halo (e.g., chloride, bromide, iodide, fluorine), methyl, alkyl (including branched and straight chain alkyl groups), alkynyl, alkenyl, methoxy, alkoxy, acetyl, amino, carboxyl, carbonyl, oxo, ester, hydroxyl, thio (e g., thiol), cyano, aryl, heteroaryl, cycloalkyl, cycloalkenyl, cycloalkylalkenyl, cycloalkylalkynyl,

cycloalkenylalkyl, cycloalkenylalkenyl, cycloalkenylalkynyl, heterocyclylalkenyl, heterocyclylalkynyl, heteroarylalkenyl, heteroarylalkynyl, arylalkenyl, arylalkynyl, heterocyclyl, spirocyclyl, heterospirocyclyl, thioalkyl (or alkylthio), arylthio, heteroarylthio, sulfone, sulfonyl, sulfoxide, amido, alkylamino, dialkylamino, arylamino, alkylarylamino, diarylamino, N-oxide, imide, enamine, imine, oxime, hydrazone, nitrile, aralkyl, cycloalkylalkyl, haloalkyl, heterocyclylalkyl, heteroaryl alkyl, nitro, thioxo, and the like. Suitable reactive groups may include, but are not necessarily limited to, azide, carboxyl, carbonyl, amine (e.g., alkyl amine (e.g., lower alkyl amine), aryl amine), halide, ester (e.g., alkyl ester (e.g., lower alkyl ester, benzyl ester), aryl ester, substituted aryl ester), cyano, thioester, thioether, sulfonyl halide, alcohol, thiol, succinimidyl ester, isothiocyanate, iodoacetamide, maleimide, hydrazine, alkynyl, alkenyl, and the like. A reactive group may facilitate covalent attachment of a molecule of interest. Suitable molecules of interest may include, but are not limited to, a detectable label; imaging agents; a toxin (including cytotoxins); a linker; a peptide; a drug (e.g., small molecule drugs); a member of a specific binding pair; an epitope tag; ligands for binding by a target receptor; tags to aid in purification; molecules that increase solubility; molecules that enhance bioavailability; molecules that increase in vivo half-life; molecules that target to a particular cell type;

molecules that target to a particular tissue; molecules that provide for crossing the blood- brain barrier; molecules to facilitate selective attachment to a surface; and the like.

Functional and reactive groups may be unsubstituted or substituted with one or more functional or reactive groups.

[00221] A cannabinoid derivative or olivetolic acid derivative may also refer to a compound lacking one or more chemical moieties found in naturally-occurring cannabinoids or olivetolic acid, yet retains the core structural features (e.g., cyclic core) of a naturally- occurring cannabinoid or olivetolic acid. Such chemical moieties may include, but are not limited to, methyl, alkyl, alkenyl, methoxy, alkoxy, acetyl, carboxyl, carbonyl, oxo, ester, hydroxyl, and the like. In some embodiments, a cannabinoid derivative or olivetolic acid derivative may also comprise one or more of any of the functional and/or reactive groups described herein. Functional and reactive groups may be unsubstituted or substituted with one or more functional or reactive groups.

[00222] The term“nucleic acid” or“nucleic acids” used herein, may refer to a polymeric form of nucleotides of any length, either ribonucleotides or deoxynucleotides. Thus, this term may include, but is not limited to, single-, double-, or multi-stranded DNA or RNA, genomic DNA, cDNA, genes, synthetic DNA or RNA, DNA-RNA hybrids, or a polymer comprising purine and pyrimidine bases or other naturally-occurring, chemically or biochemically modified, non- naturally-occurring, or derivatized nucleotide bases.

[00223] The terms“peptide,”“polypeptide,” and“protein” may be used

interchangeably herein, and may refer to a polymeric form of amino acids of any length, which can include coded and non-coded amino acids and chemically or biochemically modified or derivatized amino acids. The polypeptides disclosed herein may include full- length polypeptides, fragments of polypeptides, truncated polypeptides, fusion polypeptides, or polypeptides having modified peptide backbones. The polypeptides disclosed herein may also be variants differing from a specifically recited“reference” polypeptide (e.g., a wild- type polypeptide) by amino acid insertions, deletions, mutations, and/or substitutions.

[00224] As used herein, the term“heterologous” may refer to what is not normally found in nature. The term“heterologous nucleotide sequence” or the term“heterologous nucleic acid” may refer to a nucleic acid or nucleotide sequence not normally found in a given cell in nature. A heterologous nucleotide sequence may be: (a) foreign to its host cell (i.e., is“exogenous” to the cell); (b) naturally found in the host cell (i.e.,“endogenous”) but present at an unnatural quantity in the cell (i.e., greater or lesser quantity than naturally found in the host cell); (c) be naturally found in the host cell but positioned outside of its natural locus; or (d) be naturally found in the host cell, but with introns removed or added. A heterologous nucleic acid may be: (a) foreign to its host cell (i.e., is“exogenous” to the cell); (b) naturally found in the host cell (i.e.,“endogenous”) but present at an unnatural quantity in the cell (i.e., greater or lesser quantity than naturally found in the host cell); or (c) be naturally found in the host cell but positioned outside of its natural locus. In some embodiments, a heterologous nucleic acid may comprise a codon-optimized nucleotide sequence. A codon-optimized nucleotide sequence may be an example of a heterologous nucleotide sequence. In some embodiments, the heterologous nucleic acids disclosed herein may comprise nucleotide sequences that encode a polypeptide disclosed herein, such as a cannabinoid synthase polypeptide, but do not comprise nucleotide sequences that do not encode the polypeptide disclosed herein (e g , vector sequences, promoters, enhancers, upstream or downstream elements). In some embodiments, the heterologous nucleic acids disclosed herein may comprise nucleotide sequences encoding a polypeptide disclosed herein, such as a cannabinoid synthase polypeptide, along with nucleotide sequences that do not encode the polypeptide disclosed herein (e.g., vector sequences, promoters, enhancers, upstream or downstream elements). In some embodiments, the nucleic acids disclosed herein are heterologous.

[00225] The term“heterologous enzyme” or“heterologous polypeptide” may refer to an enzyme or polypeptide that is not normally found in a given cell in nature. The term encompasses an enzyme or polypeptide that is: (a) exogenous to a given cell (i.e., encoded by a nucleic acid that is not naturally present in the host cell or not naturally present in a given context in the host cell); or (b) naturally found in the host cell (e.g., the enzyme or polypeptide is encoded by a nucleic acid that is endogenous to the cell) but that is produced in an unnatural amount (e.g., greater or lesser than that naturally found) in the host cell. For example, a heterologous polypeptide may include a mutated version of a polypeptide naturally occurring in a host cell.

[00226] As used herein, the term“one or more heterologous nucleic acids” or“one or more heterologous nucleotide sequences” may refer to heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more polypeptides. In some embodiments, the one or more heterologous nucleic acids may comprise a nucleotide sequence encoding one polypeptide. In other embodiments, the one or more heterologous nucleic acids may comprise nucleotide sequences encoding more than one polypeptide. In certain such embodiments, the nucleotide sequences encoding the more than one polypeptide may be present on the same heterologous nucleic acid or on different heterologous nucleic acids, or combinations thereof. In some embodiments, the one or more heterologous nucleic acids may comprise nucleotide sequences encoding multiple copies of the same polypeptide. In certain such embodiments, the nucleotide sequences encoding the multiple copies of the same polypeptide may be present on the same heterologous nucleic acid or on different heterologous nucleic acids, or combinations thereof. In some embodiments, the one or more heterologous nucleic acids may comprise nucleotide sequences encoding multiple copies of different polypeptides. In certain such embodiments, the nucleotide sequences encoding the multiple copies of the different polypeptides may be present on the same heterologous nucleic acid or on different heterologous nucleic acids, or combinations thereof.

[00227] “Operably linked” may refer to an arrangement of elements wherein the components so described are configured so as to perform their usual function. Thus, control sequences operably linked to a coding sequence are capable of effecting the expression of the coding sequence. The control sequences need not be contiguous with the coding sequence, so long as they function to direct the expression thereof. Thus, for example, intervening untranslated yet transcribed sequences can be present between a promoter sequence and the coding sequence and the promoter sequence can still be considered “operably linked” to the coding sequence.

[00228] ‘Isolated” may refer to polypeptides or nucleic acids that are substantially or essentially free from components that normally accompany them in their natural state.

An isolated polypeptide or nucleic acid may be other than in the form or setting in which it is found in nature. Isolated polypeptides and nucleic acids therefore may be distinguished from the polypeptides and nucleic acids as they exist in natural cells. An isolated nucleic acid or polypeptide may be purified from one or more other components in a mixture with the isolated nucleic acid or polypeptide, if such components are present. [00229] A“modified host cell” (also may be referred to as a“recombinant host cell”) may refer to a host cell into which has been introduced a nucleic acid (e.g., a heterologous nucleic acid), e.g., an expression vector or construct. For example, a modified eukaryotic host cell may be produced through introduction into a suitable eukaryotic host cell of a nucleic acid (e.g., a heterologous nucleic acid).

[00230] As used herein, a“cell-free system” may refer to a cell lysate, cell extract or other preparation in which substantially all of the cells in the preparation have been disrupted or otherwise processed so that all or selected cellular components, e.g., organelles, proteins, nucleic acids, the cell membrane itself (or fragments or components thereof), or the like, are released from the cell or resuspended into an appropriate medium and/or purified from the cellular milieu. Cell-free systems can include reaction mixtures prepared from purified and/or isolated polypeptides and suitable reagents and buffers.

[00231] In some embodiments, conservative substitutions may be made in the amino acid sequence of a polypeptide without disrupting the three-dimensional structure or function of the polypeptide. Conservative substitutions may be accomplished by the skilled artisan by substituting amino acids with similar hydrophobicity, polarity, and R-chain length for one another. Additionally, by comparing aligned sequences of homologous proteins from different species, conservative substitutions may be identified by locating amino acid residues that have been mutated between species without altering the basic functions of the encoded proteins. The term“conservative amino acid substitution” may refer to the interchangeability in proteins of amino acid residues having similar side chains. For example, a group of amino acids having aliphatic side chains may consist of glycine, alanine, valine, leucine, and isoleucine; a group of amino acids having aliphatic-hydroxyl side chains may consist of serine and threonine; a group of amino acids having amide containing side chains may consist of asparagine and glutamine; a group of amino acids having aromatic side chains may consist of phenylalanine, tyrosine, and tryptophan; a group of amino acids having basic side chains may consist of lysine, arginine, and histidine; a group of amino acids having acidic side chains may consist of glutamate and aspartate; and a group of amino acids having sulfur containing side chains may consist of cysteine and methionine.

Exemplary conservative amino acid substitution groups are: valine-leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine-valine, and asparagine-glutamine.

[00232] A polynucleotide or polypeptide has a certain percent“sequence identity” to another polynucleotide or polypeptide, meaning that, when aligned, that percentage of bases or amino acids are the same, and in the same relative position, when comparing the two sequences. Sequence identity can be determined in a number of different manners. To determine sequence identity, sequences can be aligned using various methods and computer programs (e.g., BLAST, T-COFFEE, MUSCLE, MAFFT, etc.), available over the world wide web at sites including ncbi.nlm.nili.gov/BLAST, ebi.ac.uk/Tools/msa/tcoffee/ebi.ac.uk/ Tools/msa/muscle/mafft.cbrc.jp/alignment/software/. See, e.g., Altschul et al. (1990), J. Mol. Biol. 215:403-10.

[00233] Before the present disclosure is further described, it is to be understood that this disclosure is not limited to particular embodiments described, as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.

[00234] Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limit of that range and any other stated or intervening value in that stated range, is encompassed within the disclosure. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges, and are also

encompassed within the disclosure, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the disclosure.

[00235] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present disclosure, the preferred methods and materials are now described. All publications mentioned herein are incorporated herein by reference to disclose and describe the methods and/or materials in connection with which the publications are cited.

[00236] It must be noted that as used herein and in the appended claims, the singular forms“a,”“an,” and“the” may include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to“a cannabinoid compound” or“cannabinoid” may include a plurality of such compounds and reference to“the modified host cell” may include reference to one or more modified host cells and equivalents thereof known to those skilled in the art, and so forth. It is further noted that the claims may be drafted to exclude any optional element. As such, this statement is intended to serve as antecedent basis for use of such exclusive terminology as“solely,”“only” and the like in connection with the recitation of claim elements, or use of a“negative” limitation. [00237] It is appreciated that certain features of the disclosure, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the disclosure, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable sub-combination. All combinations of the embodiments pertaining to the disclosure are specifically embraced by the present disclosure and are disclosed herein just as if each and every combination was individually and explicitly disclosed. In addition, all sub combinations of the various embodiments and elements thereof are also specifically embraced by the present disclosure and are disclosed herein just as if each and every such sub-combination was individually and explicitly disclosed herein.

Codon-Optimized Variants of Nucleotide Sequences Encoding Cannabinoid Synthase Polypeptides

[00238] Herein, nucleic acids comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide are disclosed. These new nucleic acids of the present disclosure can be used to modify host cells to express cannabinoid synthase polypeptides. In turn, these modified host cells can be useful for producing cannabinoids and cannabinoid derivatives. Surprisingly, expression of the cannabinoid synthase polypeptides in a host cell modified with one or more nucleic acids comprising a codon- optimized nucleotide sequence encoding a cannabinoid synthase polypeptide did not significantly decrease the growth or viability of the modified host cell compared to an unmodified host cell. In some embodiments, a culture of modified host cells of the disclosure comprising one or more nucleic acids comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide has a cell density that is at least 25% or greater, at least 30% or greater, at least 35% or greater, at least 40% or greater, at least 45% or greater, at least 50% or greater, at least 55% or greater, at least 60% or greater, at least 65% or greater, at least 70% or greater, at least 75% or greater, at least 80% or greater, at least 85% or greater at least 90% or greater, at least 95% or greater, at least 100% or greater, at least 110% or greater, at least 120% or greater, at least 130% or greater, at least 140% or greater, or at least 150% or greater than the cell density of a culture of unmodified control host cells grown for the same period, in the same culture medium, and under the same culture conditions. Commonly, expression of cannabinoid synthase polypeptides by host cells modified to express these polypeptides is toxic to the host cell, reducing viability and resulting in cell death. Aggregation of the cannabinoid synthase polypeptides within the modified host cell is often observed and can be toxic to the modified host cell.

[00239] Without wishing to be bound by theory, slowing translation of the mRNA resulting from the codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide in certain regions may improve expression yields and activity of the cannabinoid synthase polypeptide and improve viability of the modified host cell. Translation of the mRNA resulting from the codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide may be optimized through use of optimal or non-optimal codons at specific regions of the mRNA. Improving recruitment of the messenger ribonucleoprotein (mRNP) to the secretory pathway may be achieved by favoring translational pause sites after the signal sequence has been translated, giving greater time for the signal recognition particle (SRP) to bind the mRNP and recruit the latter to the ER— thus disfavoring runaway cytoplasmic translation and misfolding.

[00240] Without wishing to be bound by theory, slowing translation of the resulting mRNA transcribed from the nucleotide sequence encoding a cannabinoid synthase polypeptide may improve translocation of the nascent cannabinoid synthase polypeptide chain to the endoplasmic reticulum (ER), assisting in delivery of the nascent cannabinoid synthase polypeptide chain to the secretory pathway. Slowing translation of the mRNA resulting from the nucleotide sequence encoding a cannabinoid synthase polypeptide through use of a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide may prevent saturation of the secretory pathway leading to possible protein degradation and/or modified host cell death or a loss of modified host cell viability.

[00241] In some embodiments, a codon-optimized nucleotide sequence may be optimized for expression in a yeast cell. In certain such embodiments, the yeast cell is Saccharomyces cerevisiae.

Nucleic Acids Comyrisine a Codon-Oytimized Nucleotide Sequence Encoding a

Cannabinoid Synthase Polwevtide

[00242] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, such as a tetrahydrocannabinolic acid synthase polypeptide, a cannabichromenic acid synthase polypeptide, or a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabichromenic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabidiolic acid synthase polypeptide.

[00243] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO: l, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence is set forth in SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO: l, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8, or a codon degenerate nucleotide sequence of any of the foregoing, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO: 8, or a codon degenerate nucleotide sequence of any of the foregoing, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO: l, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO: l, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID N0:7, or SEQ ID NO:8, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00244] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO: l, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO: l, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO: 6, SEQ ID NO: 7, or SEQ ID NO: 8, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO: l, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide

[00245] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO: 8, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO: 8, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO: 8, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO: 8, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00246] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO: l, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence is set forth in SEQ ID NO: 1, or a codon degenerate nucleotide sequence thereof, wherein the cannabinoid synthase polypeptide is a

cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO: 1, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO: 1, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO: l, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00247] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO: 1, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO: 1, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO: 1, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO: 1, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00248] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO:2, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence is set forth in SEQ ID NO:2, or a codon degenerate nucleotide sequence thereof, wherein the cannabinoid synthase polypeptide is a

cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:2, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:2, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:2, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00249] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:2, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:2, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:2, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:2, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00250] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO:3, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence is set forth in SEQ ID NO:3, or a codon degenerate nucleotide sequence thereof, wherein the cannabinoid synthase polypeptide is a

cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:3, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:3, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:3, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00251] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:3, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:3, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:3, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:3, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00252] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO:4, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence is set forth in SEQ ID NO:4, or a codon degenerate nucleotide sequence thereof, wherein the cannabinoid synthase polypeptide is a

cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:4, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:4, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:4, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00253] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:4, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:4, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:4, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:4, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00254] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO:5, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence is set forth in SEQ ID NO:5, or a codon degenerate nucleotide sequence thereof, wherein the cannabinoid synthase polypeptide is a

cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:5, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:5, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:5, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00255] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO: 5, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO: 5, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO: 5, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO: 5, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00256] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO:6, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence is set forth in SEQ ID NO: 6, or a codon degenerate nucleotide sequence thereof, wherein the cannabinoid synthase polypeptide is a

cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:6, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:6, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:6, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00257] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:6, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:6, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:6, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:6, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00258] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO:7, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence is set forth in SEQ ID NO: 7, or a codon degenerate nucleotide sequence thereof, wherein the cannabinoid synthase polypeptide is a

cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:7, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:7, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:7, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00259] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:7, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:7, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:7, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:7, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00260] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO: 8, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence is set forth in SEQ ID NO: 8, or a codon degenerate nucleotide sequence thereof, wherein the cannabinoid synthase polypeptide is a

cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:8, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:8, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:8, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00261] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO: 8, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO: 8, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO: 8, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO: 8, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00262] The disclosure provides nucleic acids comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide lacking a stop codon. Nucleic acids comprising a codon-optimized nucleotide sequence encoding cannabinoid synthase polypeptides lacking the stop codon may be useful for expressing said polypeptides in a construct comprising T2A elements. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO:256 (corresponding to SEQ ID NO: 1 but lacking the stop codon), SEQ ID NO:258 (corresponding to SEQ ID NO: 2 but lacking the stop codon), SEQ ID NO: 259

(corresponding to SEQ ID NO: 3 but lacking the stop codon), SEQ ID NO: 260

(corresponding to SEQ ID NO: 4 but lacking the stop codon), SEQ ID NO: 261

(corresponding to SEQ ID NO: 5 but lacking the stop codon), SEQ ID NO: 262

(corresponding to SEQ ID NO: 7 but lacking the stop codon), or SEQ ID NO: 263

(corresponding to SEQ ID NO: 8 but lacking the stop codon), wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO:263, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO:256, SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO: 260, SEQ ID NO: 261, SEQ ID NO: 262, or SEQ ID NO:263, or a codon degenerate nucleotide sequence of any of the foregoing, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO:263, or a codon degenerate nucleotide sequence of any of the foregoing, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:256, SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO:263, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:261, SEQ ID NO:262, or SEQ ID NO:263, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:256, SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO:263, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:258, SEQ ID N0 259, SEQ ID NO 260, SEQ ID N0 261, SEQ ID N0 262, or SEQ ID N0 263, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:256, SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO:263, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:258, SEQ ID NO:259,

SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO:263, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00263] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:256, SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO 260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO:263, wherein the cannabinoid synthase polypeptide is a

cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:256, SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO:263, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:256, SEQ ID NO:258, SEQ ID N0 259, SEQ ID NO 260, SEQ ID N0 261, SEQ ID N0 262, or SEQ ID N0 263, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:256, SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO:263, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00264] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID N0 261, SEQ ID NO:262, or SEQ ID NO:263, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID N0 261, SEQ ID NO:262, or SEQ ID NO:263, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID N0 261, SEQ ID NO:262, or SEQ ID NO:263, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID N0 261, SEQ ID NO:262, or SEQ ID NO:263, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00265] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO:256, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence is set forth in SEQ ID NO:256, or a codon degenerate nucleotide sequence thereof, wherein the cannabinoid synthase polypeptide is a

cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:256, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:256, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:256, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00266] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:256, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:256, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:256, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:256, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00267] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO:258, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence is set forth in SEQ ID NO:258, or a codon degenerate nucleotide sequence thereof, wherein the cannabinoid synthase polypeptide is a

cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:258, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:258, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:258, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00268] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:258, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:258, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:258, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:258, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00269] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO:259, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence is set forth in SEQ ID NO:259, or a codon degenerate nucleotide sequence thereof, wherein the cannabinoid synthase polypeptide is a

cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:259, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:259, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:259, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00270] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:259, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:259, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:259, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:259, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00271] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO:260, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence is set forth in SEQ ID NO:260, or a codon degenerate nucleotide sequence thereof, wherein the cannabinoid synthase polypeptide is a

cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:260, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:260, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:260, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00272] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:260, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:260, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:260, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:260, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00273] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO:26l, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence is set forth in SEQ ID NO:26l, or a codon degenerate nucleotide sequence thereof, wherein the cannabinoid synthase polypeptide is a

cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:26l, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:26l, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:26l, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00274] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:26l, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:26l, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:26l, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:26l, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00275] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO:262, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence is set forth in SEQ ID NO:262, or a codon degenerate nucleotide sequence thereof, wherein the cannabinoid synthase polypeptide is a

cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:262, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:262, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:262, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00276] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:262, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:262, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:262, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:262, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00277] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO:263, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence is set forth in SEQ ID NO:263, or a codon degenerate nucleotide sequence thereof, wherein the cannabinoid synthase polypeptide is a

cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:263, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:263, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:263, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00278] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:263, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:263, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:263, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:263, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00279] The disclosure provides nucleic acids comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide with an N-terminal truncation to remove the native signal sequence polypeptide. Nucleic acids comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide with an N-terminal truncation to remove the native signal sequence polypeptide may be useful for expressing cannabinoid synthase polypeptides comprising a signal sequence polypeptide different than that of the native signal sequence polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO:242 or SEQ ID NO:244, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide with an N- terminal truncation to remove the native signal sequence polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO: 242 or SEQ ID NO: 244, or a codon degenerate nucleotide sequence of any of the foregoing, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:242 or SEQ ID NO:244, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:242 or SEQ ID NO:244, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:242 or SEQ ID NO:244, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide.

[00280] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:242 or SEQ ID NO:244, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide with an N-terminal truncation to remove the native signal sequence polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:242 or SEQ ID NO:244, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:242 or SEQ ID NO:244, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:242 or SEQ ID NO:244, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide.

[00281] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO:242, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide with an N-terminal truncation to remove the native signal sequence polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence is set forth in SEQ ID NO:242, or a codon degenerate nucleotide sequence thereof, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:242, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:242, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:242, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide.

[00282] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:242, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide with an N-terminal truncation to remove the native signal sequence polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:242, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:242, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:242, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide.

[00283] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO:244, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide with an N-terminal truncation to remove the native signal sequence polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence is set forth in SEQ ID NO:244, or a codon degenerate nucleotide sequence thereof, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:244, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:244, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:244, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide.

[00284] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:244, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide with an N-terminal truncation to remove the native signal sequence polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:244, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:244, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:244, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide.

[00285] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO:389, SEQ ID NO:391,

SEQ ID NO:393, SEQ ID NO:395, SEQ ID NO: 397, SEQ ID NO :399, SEQ ID NO:401, SEQ ID NO:403, SEQ ID NO:405, SEQ ID NO:407, SEQ ID NO :409, SEQ ID NO:4l 1, SEQ ID NO:413, SEQ ID NO:415, SEQ ID NO :417, SEQ ID NO :4l9, SEQ ID NO:421, SEQ ID NO:423, SEQ ID NO:425, SEQ ID NO:427, SEQ ID NO :429, SEQ ID NO: 431, SEQ ID NO:433, SEQ ID NO:435, SEQ ID NO:437, SEQ ID NO :439, SEQ ID NO:441, SEQ ID NO:443, SEQ ID NO:445, SEQ ID NO:447, SEQ ID NO :449, SEQ ID NO:451, SEQ ID NO:453, SEQ ID NO:455, SEQ ID NO:457, SEQ ID NO :459, SEQ ID NO:461, SEQ ID NO:463, SEQ ID NO:465, SEQ ID NO:467, SEQ ID NO :469, SEQ ID NO:471, SEQ ID NO:473, SEQ ID NO:475, SEQ ID NO:477, SEQ ID NO :479, SEQ ID NO:481, SEQ ID NO:483, SEQ ID NO:485, SEQ ID NO:487, SEQ ID NO :489, SEQ ID NO:491, SEQ ID NO:493, SEQ ID NO:495, SEQ ID NO 497, SEQ ID NO :499, SEQ ID NO 501, SEQ ID NO:503, SEQ ID NO:505, SEQ ID NO: 507, SEQ ID NO :509, SEQ ID NO:5l 1, SEQ ID NO:513, SEQ ID NO:515, SEQ ID NO:517, SEQ ID NO :5l9, SEQ ID NO:521, SEQ ID NO:523, SEQ ID N0 525, SEQ ID NO 527, SEQ ID NO 529, SEQ ID NO: 531, SEQ ID NO:533, SEQ ID NO:535, SEQ ID NO:537, SEQ ID NO :539, SEQ ID NO:541, SEQ ID NO:543, SEQ ID NO:545, SEQ ID NO: 547, SEQ ID NO :549, SEQ ID NO:551, SEQ ID NO:553, SEQ ID NO:555, SEQ ID NO:557, SEQ ID NO :559, SEQ ID NO:561, SEQ ID NO:563, SEQ ID NO:565, SEQ ID NO: 567, SEQ ID NO :569, SEQ ID NO: 571, SEQ ID NO:573, SEQ ID NO:579, SEQ ID NO:58l, SEQ ID NO :583, SEQ ID NO:585, SEQ ID NO: 587, SEQ ID NO: 589, SEQ ID NO:59l, SEQ ID NO :593, SEQ ID NO:595, SEQ ID NO: 597, or SEQ ID NO:599. In certain such embodiments, the cannabinoid synthase polypeptide is a variant cannabidiolic acid synthase polypeptide.

[00286] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO:389, SEQ ID NO:391, SEQ ID NO:393, SEQID NO:395, SEQID NO:397, SEQID NO:399, SEQ ID NO:401, SEQ ID NO:403, SEQID NO:405, SEQID NO:407, SEQID NO:409, SEQ ID NO:411, SEQ ID NO:413, SEQID NO:415, SEQID NO:417, SEQID NO:419, SEQ ID NO:421, SEQ ID NO:423, SEQID NO:425, SEQID NO:427, SEQID NO:429, SEQ ID NO:431, SEQ ID NO:433, SEQID NO:435, SEQID NO:437, SEQID NO:439, SEQ ID NO:441, SEQ ID NO:443, SEQID NO:445, SEQID NO:447, SEQID NO:449, SEQ ID NO:451, SEQ ID NO:453, SEQID NO:455, SEQID NO:457, SEQID NO:459, SEQ ID NO:461, SEQ ID NO:463, SEQID NO:465, SEQID NO:467, SEQID NO:469, SEQ ID NO:471, SEQ ID NO:473, SEQID NO:475, SEQID NO:477, SEQID NO:479, SEQ ID NO:481, SEQ ID NO:483, SEQID NO:485, SEQID NO:487, SEQID NO:489, SEQ ID NO:491, SEQ ID NO:493, SEQID NO:495, SEQID NO:497, SEQID NO:499, SEQ ID NO:501, SEQ ID NO:503, SEQID NO:505, SEQID NO:507, SEQID NO:509, SEQ ID NO:511, SEQ ID NO:513, SEQID NO:515, SEQID NO:517, SEQID NO:519, SEQ ID NO:521, SEQ ID NO:523, SEQID NO:525, SEQID NO:527, SEQID NO: 529, SEQ ID NO:531, SEQ ID NO:533, SEQID NO:535, SEQID NO:537, SEQID NO:539, SEQ ID NO:541, SEQ ID NO:543, SEQID NO:545, SEQID NO:547, SEQID NO: 549, SEQ ID NO:551, SEQ ID NO:553, SEQID NO:555, SEQID NO:557, SEQID NO:559, SEQ ID NO:561, SEQ ID NO:563, SEQID NO:565, SEQID NO:567, SEQID NO:569, SEQ ID NO:571, SEQ ID NO:573, SEQID NO:579, SEQID NO:581, SEQID NO:583, SEQ ID NO:585, SEQ ID NO: 587, SEQID NO:589, SEQID NO:591, SEQID NO:593, SEQ ID N0595, SEQ ID NO: 597, or SEQ ID NO: 599, or a codon degenerate nucleotide sequence of any of the foregoing. In certain such embodiments, the cannabinoid synthase polypeptide is a variant cannabidiolic acid synthase polypeptide.

[00287] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO:606, SEQ ID NO:608, SEQ ID NO:610, SEQIDNO:612, SEQIDNO:614, SEQIDNO:616, SEQIDNO:618, SEQ ID NO:620, SEQ ID NO:622, SEQ ID NO:624, SEQ ID NO:626, SEQ ID NO:628, SEQ ID NO:630, SEQ ID NO:632, SEQ ID NO:634, SEQ ID NO:636, SEQ ID NO:638, SEQ ID NO:640, SEQ ID NO:642, SEQ ID NO:644, SEQ ID NO:646, SEQ ID NO:648, SEQ ID NO:650, SEQ ID NO:652, SEQ ID NO:654, SEQ ID NO:656, SEQ ID NO:658, SEQ ID NO:660, SEQ ID NO:662, SEQ ID NO:664, SEQ ID NO:666, SEQ ID NO:668, SEQ ID NO:670, SEQ ID NO:672, SEQ ID NO:674, SEQ ID NO:676, SEQ ID NO:678, SEQ ID NO:680, SEQ ID NO:682, SEQ ID NO:684, SEQ ID NO:686, SEQ ID NO:688, SEQ ID NO:690, SEQ ID NO:692, SEQ ID NO:694, SEQ ID NO:696, SEQ ID NO:698,

SEQ ID NO:700, SEQ ID NO:702, SEQ ID NO:704, SEQ ID NO:706, SEQ ID NO:708,

SEQ ID NO:710, SEQ ID NO:712, SEQ ID NO:714, SEQ ID NO:7l6, SEQ ID NO:7l8, SEQ ID NO:720, SEQ ID NO:722, SEQ ID NO:724, SEQ ID NO:726, SEQ ID NO:728,

SEQ ID NO:730, SEQ ID NO:732, SEQ ID NO:734, SEQ ID NO:736, SEQ ID NO:738,

SEQ ID NO:740, or SEQ ID NO:742. In certain such embodiments, the cannabinoid synthase polypeptide is a variant tetrahydrocannabinolic acid synthase polypeptide.

[00288] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO:606, SEQ ID NO:608, SEQ ID NO:610, SEQ ID NO:612, SEQ ID NO:614, SEQ ID NO:6l6, SEQ ID NO:6l8, SEQ ID NO:620, SEQ ID NO:622, SEQ ID NO:624, SEQ ID NO:626, SEQ ID NO:628,

SEQ ID NO:630, SEQ ID NO:632, SEQ ID NO:634, SEQ ID NO:636, SEQ ID NO:638,

SEQ ID NO:640, SEQ ID NO:642, SEQ ID NO:644, SEQ ID NO:646, SEQ ID NO:648,

SEQ ID NO:650, SEQ ID NO:652, SEQ ID NO:654, SEQ ID NO:656, SEQ ID NO:658,

SEQ ID NO:660, SEQ ID NO:662, SEQ ID NO:664, SEQ ID NO:666, SEQ ID NO:668,

SEQ ID NO:670, SEQ ID NO:672, SEQ ID NO:674, SEQ ID NO:676, SEQ ID NO:678,

SEQ ID NO:680, SEQ ID NO:682, SEQ ID NO:684, SEQ ID NO:686, SEQ ID NO:688,

SEQ ID NO:690, SEQ ID NO:692, SEQ ID NO:694, SEQ ID NO:696, SEQ ID NO:698,

SEQ ID NO:700, SEQ ID NO:702, SEQ ID NO:704, SEQ ID NO:706, SEQ ID NO:708,

SEQ ID NO 730, SEQ ID N0 732, SEQ ID N0 734, SEQ ID N0 736, SEQ ID N0 738,

SEQ ID NO:740, or SEQ ID NO:742, or a codon degenerate nucleotide sequence of any of the foregoing. In certain such embodiments, the cannabinoid synthase polypeptide is a variant tetrahydrocannabinolic acid synthase polypeptide.

[00289] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO:238, SEQ ID NO:271, SEQ ID NO:272, SEQ ID NO:273, or SEQ ID NO:274, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO:238, SEQ ID NO:27l, SEQ ID NO:272, SEQ ID NO:273, or SEQ ID NO:274, or a codon degenerate nucleotide sequence of any of the foregoing, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:238, SEQ ID NO:271, SEQ ID NO:272, SEQ ID NO:273, or SEQ ID NO:274, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:238, SEQ ID NO:271, SEQ ID NO:272, SEQ ID NO:273, or SEQ ID NO:274, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:238, SEQ ID NO:271,

SEQ ID NO:272, SEQ ID NO:273, or SEQ ID NO:274, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide.

[00290] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:238, SEQ ID NO:27l, SEQ ID N0 272, SEQ ID N0 273, or SEQ ID NO:274, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:238, SEQ ID NO:271, SEQ ID NO:272, SEQ ID NO:273, or SEQ ID NO:274, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:238, SEQ ID NO:271, SEQ ID NO:272, SEQ ID NO:273, or SEQ ID NO:274, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:238, SEQ ID NO:271, SEQ ID NO:272, SEQ ID NO:273, or SEQ ID NO:274, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide.

[00291] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO:238, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence is set forth in SEQ ID NO:238, or a codon degenerate nucleotide sequence thereof, wherein the cannabinoid synthase polypeptide is a

tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:238, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:238, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:238, wherein the cannabinoid synthase polypeptide is a

tetrahydrocannabinolic acid synthase polypeptide.

[00292] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:238, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:238, wherein the cannabinoid synthase polypeptide is a

tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:238, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:238, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide.

[00293] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO:27l, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence is set forth in SEQ ID NO:27l, or a codon degenerate nucleotide sequence thereof, wherein the cannabinoid synthase polypeptide is a

tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:271, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:271, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:27l, wherein the cannabinoid synthase polypeptide is a

tetrahydrocannabinolic acid synthase polypeptide.

[00294] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:27l, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:27l, wherein the cannabinoid synthase polypeptide is a

tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:27l, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:27l, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide.

[00295] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO:272, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence is set forth in SEQ ID NO:272, or a codon degenerate nucleotide sequence thereof, wherein the cannabinoid synthase polypeptide is a

tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:272, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:272, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:272, wherein the cannabinoid synthase polypeptide is a

tetrahydrocannabinolic acid synthase polypeptide.

[00296] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:272, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:272, wherein the cannabinoid synthase polypeptide is a

tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:272, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:272, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide.

[00297] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO:273, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence is set forth in SEQ ID NO:273, or a codon degenerate nucleotide sequence thereof, wherein the cannabinoid synthase polypeptide is a

tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:273, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:273, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:273, wherein the cannabinoid synthase polypeptide is a

tetrahydrocannabinolic acid synthase polypeptide.

[00298] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:273, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:273, wherein the cannabinoid synthase polypeptide is a

tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:273, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:273, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide.

[00299] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO:274, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence is set forth in SEQ ID NO:274, or a codon degenerate nucleotide sequence thereof, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:274, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:274, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:274, wherein the cannabinoid synthase polypeptide is a

tetrahydrocannabinolic acid synthase polypeptide.

[00300] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:274, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:274, wherein the cannabinoid synthase polypeptide is a

tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:274, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:274, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide.

[00301] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence is set forth in SEQ ID NO:240, wherein the cannabinoid synthase polypeptide is a cannabichromenic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence is set forth in SEQ ID NO:240, or a codon degenerate nucleotide sequence thereof, wherein the cannabinoid synthase polypeptide is a

cannabichromenic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:240, wherein the cannabinoid synthase polypeptide is a cannabichromenic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:240, wherein the cannabinoid synthase polypeptide is a cannabichromenic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:240, wherein the cannabinoid synthase polypeptide is a cannabichromenic acid synthase polypeptide.

[00302] Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:240, wherein the cannabinoid synthase polypeptide is a cannabichromenic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:240, wherein the cannabinoid synthase polypeptide is a cannabichromenic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:240, wherein the cannabinoid synthase polypeptide is a cannabichromenic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:240, wherein the cannabinoid synthase polypeptide is a cannabichromenic acid synthase polypeptide.

[00303] The disclosure provides for nucleic acids comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide and a nucleotide sequence encoding a signal sequence polypeptide. In certain such embodiments, after transcription and translation, the resulting cannabinoid synthase polypeptide is modified with the signal sequence polypeptide. In some embodiments, the signal sequence polypeptide is a secretory signal sequence polypeptide. In some embodiments, the secretory signal sequence polypeptide is a native secretory signal sequence polypeptide. In some embodiments, the secretory signal sequence polypeptide is a synthetic secretory signal sequence polypeptide.

In some embodiments, the secretory signal sequence polypeptide is an endoplasmic reticulum retention signal sequence polypeptide. In certain such embodiments, the endoplasmic reticulum retention signal sequence polypeptide is a HDEL polypeptide or a KDEL polypeptide. In some embodiments, the secretory signal sequence polypeptide is a vacuolar localization signal sequence polypeptide. In certain such embodiments, the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide or a PRClt polypeptide. In certain such embodiments, the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide. In some embodiments, the secretory signal sequence polypeptide is a plasma membrane localization signal sequence polypeptide. In some embodiments, the secretory signal sequence polypeptide is a mitochondrial targeting signal sequence polypeptide. In some embodiments, the secretory signal sequence polypeptide is a Golgi targeting signal sequence polypeptide. In some embodiments, the secretory signal sequence polypeptide is a peroxisome targeting signal sequence polypeptide. In some embodiments, the peroxisome targeting signal sequence polypeptide is a PEX8 polypeptide. In some embodiments, the secretory signal sequence polypeptide is a mating factor secretory signal sequence polypeptide (e.g., a MF polypeptide or an evolved MF polypeptide (MFev)).

[00304] In some embodiments of the nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide and a nucleotide sequence encoding a signal sequence polypeptide, the codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide is as set forth in SEQ ID NO:242, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide. In some embodiments of the nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide and a nucleotide sequence encoding a signal sequence polypeptide, the codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide is as set forth in SEQ ID NO:242, or a codon degenerate nucleotide sequence thereof, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide. In some embodiments of the nucleic acid comprising a codon- optimized nucleotide sequence encoding a cannabinoid synthase polypeptide and a nucleotide sequence encoding a signal sequence polypeptide, the codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:242, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide. In some embodiments of the nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide and a nucleotide sequence encoding a signal sequence polypeptide, the codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide have at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:242, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide. In some embodiments of the nucleic acid comprising a codon- optimized nucleotide sequence encoding a cannabinoid synthase polypeptide and a nucleotide sequence encoding a signal sequence polypeptide, the codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide have at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:242, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide. In some embodiments, the signal sequence polypeptide is a secretory signal sequence polypeptide. In certain such embodiments, the secretory signal sequence polypeptide is a vacuolar localization signal sequence polypeptide, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide.

[00305] Some embodiments of the disclosure relate to a nucleic acid comprising a nucleotide sequence encoding a signal sequence polypeptide and a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:242, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a nucleotide sequence encoding a signal sequence polypeptide and a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:242, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a nucleotide sequence encoding a signal sequence polypeptide and a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:242, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a nucleotide sequence encoding a signal sequence polypeptide and a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:242, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide. In some embodiments, the signal sequence polypeptide is a secretory signal sequence polypeptide. In certain such embodiments, the secretory signal sequence polypeptide is a vacuolar localization signal sequence polypeptide, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide.

[00306] In some embodiments of the nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide and a nucleotide sequence encoding a signal sequence polypeptide, the codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide is as set forth in SEQ ID NO:244, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide. In some embodiments of the nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide and a nucleotide sequence encoding a signal sequence polypeptide, the codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide is as set forth in SEQ ID NO:244, or a codon degenerate nucleotide sequence thereof, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide. In some embodiments of the nucleic acid comprising a codon- optimized nucleotide sequence encoding a cannabinoid synthase polypeptide and a nucleotide sequence encoding a signal sequence polypeptide, the codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:244, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide. In some embodiments of the nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide and a nucleotide sequence encoding a signal sequence polypeptide, the codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:244, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide. In some embodiments of the nucleic acid comprising a codon- optimized nucleotide sequence encoding a cannabinoid synthase polypeptide and a nucleotide sequence encoding a signal sequence polypeptide, the codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:244, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide. In some embodiments, the signal sequence polypeptide is a secretory signal sequence polypeptide. In certain such embodiments, the secretory signal sequence polypeptide is a vacuolar localization signal sequence polypeptide, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide.

[00307] Some embodiments of the disclosure relate to a nucleic acid comprising a nucleotide sequence encoding a signal sequence polypeptide and a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:244, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a nucleotide sequence encoding a signal sequence polypeptide and a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:244, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a nucleotide sequence encoding a signal sequence polypeptide and a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:244, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide. Some embodiments of the disclosure relate to a nucleic acid comprising a nucleotide sequence encoding a signal sequence polypeptide and a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the codon- optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:244, wherein the cannabinoid synthase polypeptide is a truncated cannabidiolic acid synthase polypeptide. In some embodiments, the signal sequence polypeptide is a secretory signal sequence polypeptide. In certain such embodiments, the secretory signal sequence polypeptide is a vacuolar localization signal sequence polypeptide, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide.

[00308] In some embodiments, the nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide comprises a nucleotide sequence encoding an AGA2t polypeptide. In certain such embodiments, after transcription and translation, the resulting cannabinoid synthase polypeptide is fused to the AGA2t polypeptide (e g., a fusion polypeptide).

[00309] In some embodiments, the nucleic acid comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide comprises a nucleotide sequence encoding a green fluorescent protein (GFP) polypeptide. In certain such embodiments, after transcription and translation, the resulting cannabinoid synthase polypeptide is fused to the GFP polypeptide (e.g., a fusion polypeptide).

[00310] In some embodiments, the nucleic acid disclosed herein is isolated. In some embodiments, the nucleic acid disclosed herein is purified. In some embodiments, the nucleic acid disclosed herein is isolated and/or purified.

[00311] Further included are nucleic acids that hybridize to the nucleic acids disclosed herein. Hybridization conditions may be stringent in that hybridization will occur if there is at least a 90%, 95%, or 97% sequence identity with the nucleotide sequence present in the nucleic acid encoding the polypeptides disclosed herein. The stringent conditions may include those used for known Southern hybridizations such as, for example, incubation overnight at 42 °C in a solution having 50% formamide, 5xSSC (150 mM NaCl, 15 mM trisodium citrate), 50 mM sodium phosphate (pH 7.6), 5><Denhardf s solution, 10% dextran sulfate, and 20 micrograms/milliliter denatured, sheared salmon sperm DNA, following by washing the hybridization support in 0.1 xSSC at about 65 °C. Other known hybridization conditions are well known and are described in Sambrook et al., Molecular Cloning: A Laboratory Manual, Third Edition, Cold Spring Harbor, N.Y. (2001).

[00312] The length of the nucleic acids disclosed herein may depend on the intended use. For example, if the intended use is as a primer or probe, for example for PCR amplification or for screening a library, the length of the nucleic acid will be less than the full length sequence, for example, 15-50 nucleotides In certain such embodiments, the primers or probes may be substantially identical to a highly conserved region of the nucleotide sequence or may be substantially identical to either the 5’ or 3’ end of the nucleotide sequence. In some cases, these primers or probes may use universal bases in some positions so as to be“substantially identical” but still provide flexibility in sequence recognition. It is of note that suitable primer and probe hybridization conditions are well known in the art.

[00313] Some embodiments of the disclosure relate to a vector comprising one or more nucleic acids disclosed herein. Some embodiments of the disclosure relate to an expression construct comprising one or more nucleic acids disclosed herein.

Modified Host Cells for Expressing Cannabinoid Synthase Polypeptides and for Producing Cannabinoids and Cannabinoid Derivatives

[00314] The present disclosure provides modified host cells comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide, such as tetrahydrocannabinolic acid synthase, cannabichromenic acid synthase, or cannabidiolic acid synthase polypeptides. In certain such embodiments, the modified host cells of the disclosure are for expressing a cannabinoid synthase polypeptide and/or for producing a cannabinoid or a cannabinoid derivative. In some embodiments, the nucleotide sequence encoding a cannabinoid synthase polypeptide is codon-optimized.

[00315] The disclosure also provides nucleic acids (e.g., heterologous nucleic acids), which can be introduced into microorganisms (e.g., modified host cells), resulting in expression or overexpression of one or more polypeptides, which can then be utilized in vitro (e.g., cell-free) or in vivo for the production of cannabinoids or cannabinoid derivatives. In some embodiments, these nucleic acids comprise a codon-optimized nucleotide sequence.

[00316] To produce cannabinoids or cannabinoid derivatives and create biosynthetic pathways within modified host cells, modified host cells comprising one or more

heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide may express or overexpress combinations of heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cannabinoid or cannabinoid precursor (e.g., geranylpyrophosphate (GPP), prenyl phosphates, olivetolic acid, or hexanoyl-CoA) biosynthesis. In some embodiments, the nucleotide sequences encoding the one or more polypeptides involved in cannabinoid or cannabinoid precursor biosynthesis are codon-optimized. In some embodiments, the nucleotide sequence encoding a cannabinoid synthase polypeptide is codon-optimized. In some embodiments, the modified host cells of the disclosure for producing cannabinoid or cannabinoid derivatives comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide comprise one or more modifications to modulate the expression of one or more secretory pathway polypeptides. The one or more modifications to modulate the expression of one or more secretory pathway polypeptides may include introducing into a host cell one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and/or deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides in a host cell. In some embodiments, a modified host cell of the present disclosure for producing cannabinoids or cannabinoid derivatives comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, resulting in expression or overexpression of the one or more secretory pathway polypeptides. In some embodiments, the nucleotide sequences encoding the one or more secretory pathway polypeptides are codon-optimized. In some embodiments, the modified host cell for producing cannabinoids or cannabinoid derivatives comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide comprises a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, reducing or eliminating the expression of the one or more secretory pathway polypeptides. In certain such embodiments, the modified host cells comprise a deletion of one or more genes encoding one or more secretory pathway polypeptides. In some embodiments, the modified host cells comprise a downregulation of one or more genes encoding one or more secretory pathway polypeptides.

[00317] In some embodiments of the modified host cell for producing cannabinoids or cannabinoid derivatives, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide, such as a secretory signal sequence polypeptide. In some embodiments of the modified host cell for producing cannabinoids or cannabinoid derivatives, the cannabinoid synthase polypeptide is a fusion polypeptide with an AGA2t polypeptide. In some embodiments of the modified host cell for producing cannabinoids or cannabinoid derivatives, the cannabinoid synthase polypeptide is a fusion polypeptide with a GFP polypeptide.

[00318] In some embodiments, culturing of a modified host cell for producing cannabinoids or cannabinoid derivatives in a culture medium provides for synthesis of the cannabinoid or the cannabinoid derivative.

[00319] To express cannabinoid synthase polypeptides, the modified host cells may express or overexpress heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide. In some embodiments, the nucleotide sequence encoding the cannabinoid synthase polypeptide is codon-optimized. In some embodiments, the modified host cells of the disclosure for expressing a cannabinoid synthase polypeptide comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide comprise one or more modifications to modulate the expression of one or more secretory pathway polypeptides. The one or more modifications to modulate the expression of one or more secretory pathway polypeptides may include introducing into a host cell one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and/or deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides in a host cell. In some embodiments, a modified host cell of the present disclosure for expressing a cannabinoid synthase polypeptide comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, resulting in expression or overexpression of the one or more secretory pathway polypeptides. In some embodiments, the nucleotide sequences encoding the one or more secretory pathway polypeptides are codon-optimized. In some embodiments, the modified host cell for expressing a cannabinoid synthase polypeptide comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide comprises a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, reducing or eliminating the expression of the one or more secretory pathway polypeptides. In certain such embodiments, the modified host cells comprise a deletion of one or more genes encoding one or more secretory pathway polypeptides. In some embodiments, the modified host cells comprise a downregulation of one or more genes encoding one or more secretory pathway polypeptides.

[00320] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cannabinoid or cannabinoid precursor biosynthesis. In some embodiments, the nucleotide sequences encoding the one or more polypeptides involved in cannabinoid or cannabinoid precursor biosynthesis are codon-optimized.

[00321] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide, such as a secretory signal sequence polypeptide. In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the cannabinoid synthase polypeptide is a fusion polypeptide with an AGA2t polypeptide. In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the cannabinoid synthase polypeptide is a fusion polypeptide with a GFP polypeptide.

Secretory Pathway Modifications

[00322] Secretory pathway polypeptides with modulated expression in the modified host cells of the disclosure may include, but are not limited to: chaperone or co-chaperone polypeptides, flavin adenine dinucleotide (FAD) synthetase polypeptides, flavin

mononucleotide (FMN) synthetase polypeptides, glycosidase polypeptides,

glycosyltransferase polypeptides, peptidyl-prolyl isomerase polypeptides, protein disulfide isomerase polypeptides, thiol oxidase polypeptides, fatty acid desaturase polypeptides, protein transport and trafficking polypeptides, signal peptidase or signal peptidase complex polypeptides, polypeptides involved in unfolded protein response (UPR), polypeptides involved in endoplasmic reticulum-associated degradation (ERAD), polypeptides involved in protein translocation into the endoplasmic reticulum, polypeptides involved in cell wall assembly, polypeptides involved in vacuolar protein sorting (including vacuolar proteinase polypeptides), polypeptides involved in lipid droplet assembly, and polypeptides involved in regulation of lipid metabolism. Expression of secretory pathway polypeptides may be modulated by introducing into a host cell one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and/or deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides in a host cell. In some embodiments, the nucleotide sequences encoding the one or more secretory pathway polypeptides are codon-optimized.

[00323] In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more of the following genes: a ROT2 gene, a MNS1 gene, an ALG12 gene, a FLD1 gene, an OPI1 gene, a PEP4 gene, a PRC1 gene, or a PRB1 gene. In some embodiments, the modified host cells of the disclosure comprise a deletion of one or more of the following genes: a ROT2 gene, a MNS1 gene, an ALG12 gene, a FLD1 gene, an OPI1 gene, a PEP4 gene, a PRC1 gene, or a PRB 1 gene. In some embodiments, the modified host cells of the disclosure comprise a downregulation of one or more of the following genes: a ROT2 gene, a MNS1 gene, an ALG12 gene, a FLD1 gene, an OPI1 gene, a PEP4 gene, a PRC1 gene, or a PRB1 gene.

[00324] The secretory pathway polypeptides and the nucleotide sequences encoding the secretory pathway polypeptides may be derived from any suitable source, for example, bacteria, yeast, fungi, algae, human, plant, or mouse. In some embodiments, the secretory pathway polypeptides and the nucleotide sequences encoding the pathway polypeptides may be derived from Pichia pastoris (now known as Komagataella phaffii ), Pichia finlandica , Pichia trehalophila, Pichia koclamae, Pichia membranaefaciens , Pichia opuntiae , Pichia thermotolerans, Pichia saliciaria, Pichia guercuum, Pichia pijperi , Pichia stiptis, Pichia methanolica , Pichia sp., Saccharomyces cerevisiae , Saccharomyces sp., Hansenula polymorpha (now known as Pichia angusta), Yarrowia lipolytica , Kluyveromyces sp., Kluyveromyces lactis, Kluyveromyces marxianus, Schizosaccharomyces pombe ,

Scheffersomyces stipites, Dekkera bruxellensis , Blastobotrys adeninivorans (formerly Arxula adeninivorans ), Candida albicans , Aspergillus nidulans , Aspergillus niger, Aspergillus oryzae, Trichoderma reesei, Chrysosporium lucknowense, Fusarium sp., Fusarium gramineum , Fusarium venenatum , Neurospora crassa , and the like. In some embodiments, the disclosure also encompasses orthologous genes encoding the secretory pathway polypeptides disclosed herein. Exemplary secretory pathway polypeptides disclosed herein may also include a full-length secretory pathway polypeptide, a fragment of a secretory pathway polypeptide, a variant of a secretory pathway polypeptide, a truncated secretory pathway polypeptide, or a fusion polypeptide that has at least one activity of a secretory pathway polypeptide. The disclosure also provides for nucleotide sequences encoding secretory pathway polypeptides, such as, a full-length secretory pathway polypeptide, a fragment of a secretory pathway polypeptide, a variant of a secretory pathway polypeptide, a truncated secretory pathway polypeptide, or a fusion polypeptide that has at least one activity of a secretory pathway polypeptide. In some embodiments, the nucleotide sequences encoding the secretory pathway polypeptides are codon-optimized.

Chaperone and Co-Chaperone Polypeptides

[00325] Modified host cells of the disclosure may comprise one or more modifications to modulate the expression of one or more chaperone or co-chaperone polypeptides. The one or more modifications to modulate the expression of one or more chaperone or co-chaperone polypeptides may include introducing into a host cell one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides and/or deletion or downregulation of one or more genes encoding one or more chaperone or co-chaperone polypeptides in a host cell. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, resulting in expression or overexpression of the one or more chaperone or co chaperone polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more chaperone or co-chaperone polypeptides, reducing or eliminating the expression of the one or more chaperone or co-chaperone polypeptides.

[00326] Exemplary chaperone or co-chaperone polypeptides disclosed herein may also include a full-length chaperone or co-chaperone polypeptide, a fragment of a chaperone or co-chaperone polypeptide, a variant of a chaperone or co-chaperone polypeptide, a truncated chaperone or co-chaperone polypeptide, or a fusion polypeptide that has at least one activity of a chaperone or co-chaperone polypeptide. [00327] Modified host cells of the disclosure may comprise one or more modifications to modulate the expression of one or more chaperone or co-chaperone polypeptides including, but not limited to, cytoplasmic chaperone or co-chaperone polypeptides (e.g., a SSA1 polypeptide, a SSB1 polypeptide, a PFD2s polypeptide, or a CNS1 polypeptide), ribosome assembly factor polypeptides (e.g., a DED1 polypeptide), a peptidyl-prolyl isomerase polypeptide (e.g., a CPR5 polypeptide or a FPR1 polypeptide), polypeptides that are members of the chaperonin containing T-complex (e.g., a TCP1 polypeptide, a CCT2 polypeptide, a CCT3 polypeptide, a CCT4 polypeptide, a CCT5 polypeptide, a CCT6 polypeptide, a CCT7 polypeptide, or a CCT8 polypeptide), and ER chaperone polypeptides (e.g., protein folding chaperone polypeptides, a KAR2 polypeptide, a LHSl polypeptide, a JEM1 polypeptide, a CNS1 polypeptide, a CNE1 polypeptide, a SCJ1 polypeptide, a ROTl polypeptide, or a SIL1 polypeptide). In some embodiments, the one or more modifications to modulate the expression of one or more chaperone or co-chaperone polypeptides may improve modified host cell viability. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a DED1 polypeptide. Improving modified host cell viability may improve the industrial fermentation process.

[00328] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, the one or more chaperone or co chaperone polypeptides are selected from the group consisting of: a KAR2 polypeptide, a JEM1 polypeptide, a LHSl polypeptide, a SIS1 polypeptide, a SSB1 polypeptide, a CNE1 polypeptide, a CNS1 polypeptide, a PFD2s polypeptide, a PFD1 polypeptide, a SSA1 polypeptide, a YDI1 polypeptide, a SIL1 polypeptide, a SCI1 polypeptide, a ROTl polypeptide, a TCP1 polypeptide, a CCT2 polypeptide, a CCT3 polypeptide, a CCT4 polypeptide, a CCT5 polypeptide, a CCT6 polypeptide, a CCT7 polypeptide, a CCT8 polypeptide, a DED1 polypeptide, a FPR1 polypeptide, and a CPR5 polypeptide.

[00329] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, the one or more chaperone or co chaperone polypeptides are selected from the group consisting of chaperone or co-chaperone polypeptides comprising the amino acid sequences set forth in SEQ ID NO:23 (a KAR2 polypeptide), SEQ ID NO:25 (a IEM1 polypeptide), SEQ ID NO:27 (a LHS1 polypeptide), SEQ ID NO:29 (a SIS1 polypeptide), SEQ ID NO: l3 (a SSB1 polypeptide), SEQ ID NO:3 l (a CNE1 polypeptide), SEQ ID NO: 17 (a CNS1 polypeptide), SEQ ID NO: 19 (a PFD2s polypeptide), SEQ ID NO:99 (a PFD1 polypeptide), SEQ ID NO: 11 (a SSA1 polypeptide), SEQ ID NO: l5 (a YDJ1 polypeptide), SEQ ID NO:33 (a SIL1 polypeptide), SEQ ID NO:35 (a SCJ1 polypeptide), SEQ ID NO:4l (a ROT1 polypeptide), SEQ ID NO:l01 (a TCP1 polypeptide), SEQ ID NO: 103 (a CCT2 polypeptide), SEQ ID NO: 105 (a CCT3

polypeptide), SEQ ID NO: 107 (a CCT4 polypeptide), SEQ ID NO: 109 (a CCT5 polypeptide), SEQ ID NO: 111 (a CCT6 polypeptide), SEQ ID NO: 113 (a CCT7

polypeptide), SEQ ID NO: 115 (a CCT8 polypeptide), SEQ ID NO:20l (a DED1 polypeptide), SEQ ID NO:49 (a FPR1 polypeptide), and SEQ ID NO:47 (a CPR5 polypeptide).

[00330] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, the one or more chaperone or co chaperone polypeptides are selected from the group consisting of chaperone or co-chaperone polypeptides comprising the amino acid sequences set forth in SEQ ID NO:23 (a KAR2 polypeptide), SEQ ID NO:25 (a JEM1 polypeptide), SEQ ID NO:27 (a LHS1 polypeptide), SEQ ID NO:29 (a SIS1 polypeptide), SEQ ID NO: l3 (a SSB1 polypeptide), SEQ ID NO:3 l (a CNE1 polypeptide), SEQ ID NO: 17 (a CNS1 polypeptide), SEQ ID NO: 19 (a PFD2s polypeptide), SEQ ID NO:99 (a PFD1 polypeptide), SEQ ID NO: 11 (a SSA1 polypeptide), SEQ ID NO: l5 (a YDJ1 polypeptide), SEQ ID NO:33 (a SIL1 polypeptide), SEQ ID NO:35 (a SCI1 polypeptide), SEQ ID NO:41 (a ROT1 polypeptide), SEQ ID NO:101 (a TCP1 polypeptide), SEQ ID NO: 103 (a CCT2 polypeptide), SEQ ID NO: 105 (a CCT3

polypeptide), SEQ ID NO: 115 (a CCT8 polypeptide), SEQ ID NO:201 (a DED1 polypeptide), SEQ ID NO:49 (a FPR1 polypeptide), and SEQ ID NO:47 (a CPR5 polypeptide), or a conservatively substituted amino acid sequence of any of the foregoing.

[00331] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, the one or more chaperone or co chaperone polypeptides are selected from the group consisting of chaperone or co-chaperone polypeptides comprising amino acid sequences having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:23 (a KAR2 polypeptide), SEQ ID NO:25 (a JEM1 polypeptide), SEQ ID NO:27 (a LHS1 polypeptide), SEQ ID NO:29 (a SIS1 polypeptide), SEQ ID NO: 13 (a SSB1 polypeptide), SEQ ID NO:31 (a CNE1 polypeptide), SEQ ID NO: 17 (a CNS1 polypeptide), SEQ ID NO: 19 (a PFD2s polypeptide), SEQ ID NO: 99 (a PFD1 polypeptide), SEQ ID NO: 11 (a SSA1 polypeptide), SEQ ID NO: 15 (a YDJ1 polypeptide), SEQ ID NO:33 (a SIL1 polypeptide), SEQ ID NO:35 (a SCI1 polypeptide), SEQ ID NO:4l (a ROT1 polypeptide), SEQ ID NO: 101 (a TCP1 polypeptide), SEQ ID NO: 103 (a CCT2 polypeptide), SEQ ID NO: 105 (a CCT3 polypeptide), SEQ ID NO: 107 (a CCT4 polypeptide), SEQ ID NO: 109 (a CCT5 polypeptide), SEQ ID NO: 111 (a CCT6 polypeptide), SEQ ID NO: 113 (a CCT7 polypeptide), SEQ ID NO: 115 (a CCT8 polypeptide), SEQ ID NO:201 (a DED1 polypeptide), SEQ ID NO:49 (a FPR1 polypeptide), and SEQ ID NO:47 (a CPR5 polypeptide).

[00332] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more chaperone or co-chaperone polypeptides, the one or more chaperone or co-chaperone polypeptides are selected from the group consisting of a KAR2 polypeptide, a IEM1 polypeptide, a LHSl polypeptide, a SIS1 polypeptide, a SSB1 polypeptide, a CNE1 polypeptide, a CNS1 polypeptide, a PFD2s polypeptide, a PFD1 polypeptide, a SSA1 polypeptide, a YDJ1 polypeptide, a SIL1 polypeptide, a SCI1 polypeptide, a ROTl polypeptide, a TCP1 polypeptide, a CCT2 polypeptide, a CCT3 polypeptide, a CCT4 polypeptide, a CCT5 polypeptide, a CCT6 polypeptide, a CCT7 polypeptide, a CCT8 polypeptide, a DED1 polypeptide, a FPR1 polypeptide, and a CPR5 polypeptide.

[00333] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more chaperone or co-chaperone polypeptides, the one or more chaperone or co-chaperone polypeptides are selected from the group consisting of chaperone or co-chaperone polypeptides comprising the amino acid sequences set forth in SEQ ID NO:23 (a KAR2 polypeptide), SEQ ID NO:25 (a JEM1 polypeptide), SEQ ID NO:27 (a LHS1 polypeptide), SEQ ID NO:29 (a SIS1 polypeptide), SEQ ID NO: l3 (a SSB1 polypeptide), SEQ ID NO:3 l (a CNE1 polypeptide), SEQ ID NO: 17 (a CNS1 polypeptide), SEQ ID NO: 19 (a PFD2s polypeptide), SEQ ID NO:99 (a PFD1 polypeptide), SEQ ID NO: 11 (a SSA1 polypeptide), SEQ ID NO: l5 (a YDJ1 polypeptide), SEQ ID NO:33 (a SIL1 polypeptide), SEQ ID NO:35 (a SCJ1 polypeptide), SEQ ID NO:4l (a ROT1 polypeptide), SEQ ID NO:l01 (a TCP1 polypeptide), SEQ ID NO: 103 (a CCT2 polypeptide), SEQ ID NO: 105 (a CCT3

[00334] In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a chaperone or co-chaperone polypeptide. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding two or more chaperone or co-chaperone polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding three or more chaperone or co-chaperone polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding four or more chaperone or co-chaperone polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding five or more chaperone or co-chaperone polypeptides.

[00335] In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more chaperone or co chaperone polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of two or more genes encoding two or more chaperone or co-chaperone polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of three or more genes encoding three or more chaperone or co-chaperone polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of four or more genes encoding four or more chaperone or co-chaperone polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of five or more genes encoding five or more chaperone or co-chaperone polypeptides. [00336] In some embodiments, the nucleotide sequences encoding the one or more chaperone or co-chaperone polypeptides are codon-optimized. In some embodiments, the nucleotide sequences encoding the one or more chaperone or co-chaperone polypeptides are a spliced form with the intron removed of the nucleotide sequences encoding the one or more chaperone or co-chaperone polypeptides. For example, the nucleotide sequence encoding the PFD2s polypeptide may be a spliced form that requires no additional splicing prior to translation.

[00337] In some embodiments, one or more chaperone or co-chaperone polypeptides are overexpressed in the modified host cell. Overexpression may be achieved by increasing the copy number of the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, e.g., through use of a high copy number expression vector (e g., a plasmid that exists at 10-40 copies or about 100 copies per cell) and/or by operably linking the nucleotide sequences encoding the one or more chaperone or co-chaperone polypeptides to a strong promoter. In some embodiments, the modified host cell has one copy of a heterologous nucleic acid comprising a nucleotide sequence encoding a chaperone or co-chaperone polypeptide. In some embodiments, the modified host cell has two copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a chaperone or co-chaperone polypeptide. In some embodiments, the modified host cell has three copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a chaperone or co-chaperone polypeptide. In some embodiments, the modified host cell has four copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a chaperone or co-chaperone polypeptide. In some embodiments, the modified host cell has five copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a chaperone or co-chaperone polypeptide. In some embodiments, the modified host cell has five or more copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a chaperone or co-chaperone polypeptide. Increased copy number of the heterologous nucleic acid and/or codon optimization of the nucleotide sequence may result in an increase in the desired polypeptide activity in the modified host cell.

[00338] Exemplary heterologous nucleic acids disclosed herein may include nucleic acids comprising a nucleotide sequence that encodes a chaperone or co-chaperone polypeptide, such as, a full-length chaperone or co-chaperone polypeptide, a fragment of a chaperone or co-chaperone polypeptide, a variant of a chaperone or co-chaperone polypeptide, a truncated chaperone or co-chaperone polypeptide, or a fusion polypeptide that has at least one activity of a chaperone or co-chaperone polypeptide. In some embodiments, the nucleotide sequence is codon-optimized.

[00339] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, the nucleotide sequences encoding the one or more chaperone or co-chaperone polypeptides are selected from the group consisting of nucleotide sequences set forth in SEQ ID NO:22 (encodes a KAR2 polypeptide), SEQ ID NO:24 (encodes a JEM1 polypeptide), SEQ ID NO:26 (encodes a LHS1 polypeptide), SEQ ID NO:28 (encodes a SIS1 polypeptide), SEQ ID NO: 12 (encodes a SSB1 polypeptide),

SEQ ID NO:30 (encodes a CNE1 polypeptide), SEQ ID NO: 16 (encodes a CNS1 polypeptide), SEQ ID NO: 18 (encodes a PFD2s polypeptide), SEQ ID NO:98 (encodes a PFD1 polypeptide), SEQ ID NO: 10 (encodes a SSA1 polypeptide), SEQ ID NO: 14 (encodes a YDJ1 polypeptide), SEQ ID NO:32 (encodes a SIL1 polypeptide), SEQ ID NO:34 (encodes a SCJ1 polypeptide), SEQ ID NO:40 (encodes a ROT1 polypeptide), SEQ ID NO: 100 (encodes a TCP1 polypeptide), SEQ ID NO: 102 (encodes a CCT2 polypeptide), SEQ ID NO: 104 (encodes a CCT3 polypeptide), SEQ ID NO: 106 (encodes a CCT4 polypeptide), SEQ ID NO: 108 (encodes a CCT5 polypeptide), SEQ ID NO: l 10 (encodes a CCT6 polypeptide), SEQ ID NO: 112 (encodes a CCT7 polypeptide), SEQ ID NO: 114 (encodes a CCT8 polypeptide), SEQ ID NO:200 (encodes a DED1 polypeptide), SEQ ID NO:48 (encodes a FPR1 polypeptide), and SEQ ID N0 46 (encodes a CPR5 polypeptide).

[00340] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, the nucleotide sequences encoding the one or more chaperone or co-chaperone polypeptides are selected from the group consisting of nucleotide sequences set forth in SEQ ID NO:22 (encodes a KAR2 polypeptide), SEQ ID NO:24 (encodes a JEM1 polypeptide), SEQ ID NO:26 (encodes a LHS1 polypeptide), SEQ ID NO:28 (encodes a SIS1 polypeptide), SEQ ID NO: 12 (encodes a SSB1 polypeptide),

SEQ ID NO:30 (encodes a CNE1 polypeptide), SEQ ID NO: 16 (encodes a CNS1 polypeptide), SEQ ID NO: 18 (encodes a PFD2s polypeptide), SEQ ID NO:98 (encodes a PFD1 polypeptide), SEQ ID NO: 10 (encodes a SSA1 polypeptide), SEQ ID NO: 14 (encodes a YDJ1 polypeptide), SEQ ID NO:32 (encodes a SIL1 polypeptide), SEQ ID NO:34 (encodes a SCJ1 polypeptide), SEQ ID NO:40 (encodes a ROT1 polypeptide), SEQ ID NO: 100 (encodes a TCP1 polypeptide), SEQ ID NO: 102 (encodes a CCT2 polypeptide), SEQ ID NO: 104 (encodes a CCT3 polypeptide), SEQ ID NO: 106 (encodes a CCT4 polypeptide), SEQ ID NO: 108 (encodes a CCT5 polypeptide), SEQ ID NO: l 10 (encodes a CCT6 polypeptide), SEQ ID NO: 112 (encodes a CCT7 polypeptide), SEQ ID NO: 114 (encodes a CCT8 polypeptide), SEQ ID NO:200 (encodes a DED1 polypeptide), SEQ ID NO:48 (encodes a FPR1 polypeptide), and SEQ ID NO:46 (encodes a CPR5 polypeptide), or a codon degenerate nucleotide sequence of any of the foregoing.

[00341] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, the nucleotide sequences encoding the one or more chaperone or co-chaperone polypeptides are selected from the group consisting of nucleotide sequences having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:22 (encodes a KAR2 polypeptide), SEQ ID NO:24 (encodes a JEM1 polypeptide), SEQ ID NO:26 (encodes a LHS1 polypeptide), SEQ ID NO:28 (encodes a SIS1 polypeptide), SEQ ID NO: 12 (encodes a SSB1 polypeptide), SEQ ID NO:30 (encodes a CNE1 polypeptide), SEQ ID NO: 16 (encodes a CNS1 polypeptide), SEQ ID NO: 18 (encodes a PFD2s polypeptide), SEQ ID NO:98 (encodes a PFD1 polypeptide), SEQ ID NO: 10 (encodes a SSA1 polypeptide), SEQ ID NO: 14 (encodes a YDJ1 polypeptide), SEQ ID NO:32 (encodes a SIL1 polypeptide), SEQ ID NO:34 (encodes a SCJ1 polypeptide), SEQ ID NO:40 (encodes a ROT1 polypeptide), SEQ ID NO: 100 (encodes a TCP1 polypeptide), SEQ ID NO: 102 (encodes a CCT2 polypeptide), SEQ ID NO: 104 (encodes a CCT3 polypeptide), SEQ ID NO: 106 (encodes a CCT4 polypeptide), SEQ ID NO: 108 (encodes a CCT5 polypeptide), SEQ ID NO: 110 (encodes a CCT6 polypeptide), SEQ ID NO: 112 (encodes a CCT7 polypeptide), SEQ ID NO: 114 (encodes a CCT8 polypeptide), SEQ ID NO:200 (encodes a DED1 polypeptide), SEQ ID NO:48 (encodes a FPR1 polypeptide), and SEQ ID NO:46 (encodes a CPR5 polypeptide).

[00342] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more chaperone or co-chaperone polypeptides, the one or more chaperone or co-chaperone polypeptides are encoded by nucleotide sequences selected from the group consisting of nucleotide sequences set forth in SEQ ID NO:22 (encodes a KAR2 polypeptide), SEQ ID NO:24 (encodes a JEM1 polypeptide), SEQ ID NO:26 (encodes a LHS1 polypeptide), SEQ ID NO:28 (encodes a SIS1 polypeptide), SEQ ID NO: 12 (encodes a SSB1 polypeptide),

SEQ ID NO:30 (encodes a CNE1 polypeptide), SEQ ID NO: 16 (encodes a CNS1 polypeptide), SEQ ID NO: 18 (encodes a PFD2s polypeptide), SEQ ID NO:98 (encodes a PFD1 polypeptide), SEQ ID NO: 10 (encodes a SSA1 polypeptide), SEQ ID NO: 14 (encodes a YDJ1 polypeptide), SEQ ID NO:32 (encodes a SIL1 polypeptide), SEQ ID NO:34 (encodes a SCJ1 polypeptide), SEQ ID NO:40 (encodes a ROT1 polypeptide), SEQ ID NO: 100 (encodes a TCP1 polypeptide), SEQ ID NO: 102 (encodes a CCT2 polypeptide),

SEQ ID NO: 104 (encodes a CCT3 polypeptide), SEQ ID NO: 106 (encodes a CCT4 polypeptide), SEQ ID NO: 108 (encodes a CCT5 polypeptide), SEQ ID NO: l 10 (encodes a CCT6 polypeptide), SEQ ID NO: 112 (encodes a CCT7 polypeptide), SEQ ID NO: 114 (encodes a CCT8 polypeptide), SEQ ID NO:200 (encodes a DED1 polypeptide), SEQ ID NO:48 (encodes a FPR1 polypeptide), and SEQ ID NO:46 (encodes a CPR5 polypeptide).

Flavin Adenine Dinucleotide (FAD) Synthetase Polypeptides

[00343] Modified host cells of the disclosure may comprise one or more modifications to modulate the expression of one or more flavin adenine dinucleotide (FAD) synthetase polypeptides. The one or more modifications to modulate the expression of one or more flavin adenine dinucleotide (FAD) synthetase polypeptides may include introducing into a host cell one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides and/or deletion or downregulation of one or more genes encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides in a host cell. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, resulting in expression or overexpression of the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, reducing or eliminating the expression of the one or more flavin adenine dinucleotide (FAD) synthetase

polypeptides. Flavin adenine dinucleotide (FAD) synthetase polypeptides catalyze adenylation of flavin mononucleotide (FMN) to form flavin adenine dinucleotide (FAD) coenzyme.

[00344] Exemplary flavin adenine dinucleotide (FAD) synthetase polypeptides disclosed herein may also include a full-length flavin adenine dinucleotide (FAD) synthetase polypeptide, a fragment of a flavin adenine dinucleotide (FAD) synthetase polypeptide, a variant of a flavin adenine dinucleotide (FAD) synthetase polypeptide, a truncated flavin adenine dinucleotide (FAD) synthetase polypeptide, or a fusion polypeptide that has at least one activity of a flavin adenine dinucleotide (FAD) synthetase polypeptide.

[00345] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides is a FAD1 polypeptide.

[00346] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides is a FAD1 polypeptide comprising the amino acid sequence set forth in SEQ ID NO:37.

[00347] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides is a FAD1 polypeptide comprising the amino acid sequence set forth in SEQ ID NO:37, or a conservatively substituted amino acid sequence thereof.

[00348] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides is a FAD1 polypeptide comprising an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:37.

[00349] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides is a FADl polypeptide. [00350] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides is a FAD1 polypeptide comprising the amino acid sequence set forth in SEQ ID NO:37.

[00351] In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a flavin adenine dinucleotide (FAD) synthetase polypeptide. In some

embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding two or more flavin adenine dinucleotide (FAD) synthetase polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding three or more flavin adenine dinucleotide (FAD) synthetase polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding four or more flavin adenine dinucleotide (FAD) synthetase polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding five or more flavin adenine dinucleotide (FAD) synthetase polypeptides.

[00352] In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of two or more genes encoding two or more flavin adenine dinucleotide (FAD) synthetase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of three or more genes encoding three or more flavin adenine dinucleotide (FAD) synthetase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of four or more genes encoding four or more flavin adenine dinucleotide (FAD) synthetase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of five or more genes encoding five or more flavin adenine dinucleotide (FAD) synthetase polypeptides. [00353] In some embodiments, the nucleotide sequences encoding the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides are codon-optimized.

[00354] In some embodiments, one or more flavin adenine dinucleotide (FAD) synthetase polypeptides are overexpressed in the modified host cell. Overexpression may be achieved by increasing the copy number of the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, e.g., through use of a high copy number expression vector (e.g., a plasmid that exists at 10-40 copies or about 100 copies per cell) and/or by operably linking the nucleotide sequences encoding the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides to a strong promoter. In some embodiments, the modified host cell has one copy of a heterologous nucleic acid comprising a nucleotide sequence encoding a flavin adenine dinucleotide (FAD) synthetase polypeptide. In some embodiments, the modified host cell has two copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a flavin adenine dinucleotide (FAD) synthetase polypeptide. In some embodiments, the modified host cell has three copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a flavin adenine dinucleotide (FAD) synthetase polypeptide. In some embodiments, the modified host cell has four copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a flavin adenine dinucleotide (FAD) synthetase polypeptide. In some embodiments, the modified host cell has five copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a flavin adenine dinucleotide (FAD) synthetase polypeptide. In some embodiments, the modified host cell has five or more copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a flavin adenine dinucleotide (FAD) synthetase polypeptide. Increased copy number of the heterologous nucleic acid and/or codon optimization of the nucleotide sequence may result in an increase in the desired polypeptide activity in the modified host cell.

[00355] Exemplary heterologous nucleic acids disclosed herein may include nucleic acids comprising a nucleotide sequence that encodes a flavin adenine dinucleotide (FAD) synthetase polypeptide, such as, a full-length flavin adenine dinucleotide (FAD) synthetase polypeptide, a fragment of a flavin adenine dinucleotide (FAD) synthetase polypeptide, a variant of a flavin adenine dinucleotide (FAD) synthetase polypeptide, a truncated flavin adenine dinucleotide (FAD) synthetase polypeptide, or a fusion polypeptide that has at least one activity of a flavin adenine dinucleotide (FAD) synthetase polypeptide. In some embodiments, the nucleotide sequence is codon-optimized. [00356] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, the nucleotide sequences encoding the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides is a nucleotide sequence encoding a FAD1 polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:36.

[00357] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, the nucleotide sequences encoding the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides is a nucleotide sequence encoding a FAD1 polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:36, or a codon degenerate nucleotide sequence thereof.

[00358] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, the nucleotide sequences encoding the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides is a nucleotide sequence encoding a FAD1 polypeptide, wherein the nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:36.

[00359] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides are encoded by a nucleotide sequence encoding a FAD1 polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:36.

Flavin Mononucleotide (FMN) Synthetase Polypeptides

[00360] Modified host cells of the disclosure may comprise one or more modifications to modulate the expression of one or more flavin mononucleotide (FMN) synthetase polypeptides. The one or more modifications to modulate the expression of one or more flavin mononucleotide (FMN) synthetase polypeptides may include introducing into a host cell one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin mononucleotide (FMN) synthetase polypeptides and/or deletion or downregulation of one or more genes encoding one or more flavin mononucleotide (FMN) synthetase polypeptides in a host cell. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin mononucleotide (FMN) synthetase polypeptides, resulting in expression or overexpression of the one or more flavin mononucleotide (FMN) synthetase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more flavin mononucleotide (FMN) synthetase polypeptides, reducing or eliminating the expression of the one or more flavin mononucleotide (FMN) synthetase polypeptides. Flavin

mononucleotide (FMN) synthetase polypeptides catalyze the phosphorylation of riboflavin to produce riboflavin monophosphate (FMN), a necessary cofactor for many enzymes.

[00361] Exemplary flavin mononucleotide (FMN) synthetase polypeptides disclosed herein may also include a full-length flavin mononucleotide (FMN) synthetase polypeptide, a fragment of a flavin mononucleotide (FMN) synthetase polypeptide, a variant of a flavin mononucleotide (FMN) synthetase polypeptide, a truncated flavin mononucleotide (FMN) synthetase polypeptide, or a fusion polypeptide that has at least one activity of a flavin mononucleotide (FMN) synthetase polypeptide.

[00362] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin mononucleotide (FMN) synthetase polypeptides, the one or more flavin mononucleotide (FMN) synthetase polypeptides is a FMN1 polypeptide.

[00363] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin mononucleotide (FMN) synthetase polypeptides, the one or more flavin mononucleotide (FMN) synthetase polypeptides is a FMN1 polypeptide comprising the amino acid sequence set forth in SEQ ID NO:39.

[00364] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin mononucleotide (FMN) synthetase polypeptides, the one or more flavin mononucleotide (FMN) synthetase polypeptides is a FMN1 polypeptide comprising the amino acid sequence set forth in SEQ ID NO:39, or a conservatively substituted amino acid sequence thereof. [00365] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin mononucleotide (FMN) synthetase polypeptides, the one or more flavin mononucleotide (FMN) synthetase polypeptides is a FMN1 polypeptide comprising an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:39.

[00366] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more flavin mononucleotide (FMN) synthetase polypeptides, the one or more flavin mononucleotide (FMN) synthetase polypeptides is a FMNl polypeptide.

[00367] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more flavin mononucleotide (FMN) synthetase polypeptides, the one or more flavin mononucleotide (FMN) synthetase polypeptides is a FMN1 polypeptide comprising the amino acid sequence set forth in SEQ ID NO :39.

[00368] In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a flavin mononucleotide (FMN) synthetase polypeptide. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin mononucleotide (FMN) synthetase polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding two or more flavin mononucleotide (FMN) synthetase polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding three or more flavin mononucleotide (FMN) synthetase polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding four or more flavin mononucleotide (FMN) synthetase polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding five or more flavin mononucleotide (FMN) synthetase polypeptides.

[00369] In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more flavin

mononucleotide (FMN) synthetase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of two or more genes encoding two or more flavin mononucleotide (FMN) synthetase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of three or more genes encoding three or more flavin mononucleotide (FMN) synthetase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of four or more genes encoding four or more flavin mononucleotide (FMN) synthetase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of five or more genes encoding five or more flavin mononucleotide (FMN) synthetase polypeptides.

[00370] In some embodiments, the nucleotide sequences encoding the one or more flavin mononucleotide (FMN) synthetase polypeptides are codon-optimized.

[00371] In some embodiments, one or more flavin mononucleotide (FMN) synthetase polypeptides are overexpressed in the modified host cell. Overexpression may be achieved by increasing the copy number of the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin mononucleotide (FMN) synthetase polypeptides, e.g., through use of a high copy number expression vector (e.g., a plasmid that exists at 10-40 copies or about 100 copies per cell) and/or by operably linking the nucleotide sequences encoding the one or more flavin mononucleotide (FMN) synthetase polypeptides to a strong promoter. In some embodiments, the modified host cell has one copy of a heterologous nucleic acid comprising a nucleotide sequence encoding a flavin

mononucleotide (FMN) synthetase polypeptide. In some embodiments, the modified host cell has two copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a flavin mononucleotide (FMN) synthetase polypeptide. In some embodiments, the modified host cell has three copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a flavin mononucleotide (FMN) synthetase polypeptide ln some embodiments, the modified host cell has four copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a flavin mononucleotide (FMN) synthetase polypeptide. In some embodiments, the modified host cell has five copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a flavin mononucleotide (FMN) synthetase polypeptide. In some embodiments, the modified host cell has five or more copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a flavin mononucleotide (FMN) synthetase polypeptide. Increased copy number of the heterologous nucleic acid and/or codon optimization of the nucleotide sequence may result in an increase in the desired polypeptide activity in the modified host cell.

[00372] Exemplary heterologous nucleic acids disclosed herein may include nucleic acids comprising a nucleotide sequence that encodes a flavin mononucleotide (FMN) synthetase polypeptide, such as, a full-length flavin mononucleotide (FMN) synthetase polypeptide, a fragment of a flavin mononucleotide (FMN) synthetase polypeptide, a variant of a flavin mononucleotide (FMN) synthetase polypeptide, a truncated flavin

mononucleotide (FMN) synthetase polypeptide, or a fusion polypeptide that has at least one activity of a flavin mononucleotide (FMN) synthetase polypeptide. In some embodiments, the nucleotide sequence is codon-optimized.

[00373] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin mononucleotide (FMN) synthetase polypeptides, the nucleotide sequences encoding the one or more flavin mononucleotide (FMN) synthetase polypeptides is a nucleotide sequence encoding a FMN1 polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:38.

[00374] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin mononucleotide (FMN) synthetase polypeptides, the nucleotide sequences encoding the one or more flavin mononucleotide (FMN) synthetase polypeptides is a nucleotide sequence encoding a FMN1 polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:38, or a codon degenerate nucleotide sequence thereof.

[00375] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin mononucleotide (FMN) synthetase polypeptides, the nucleotide sequences encoding the one or more flavin mononucleotide (FMN) synthetase polypeptides is a nucleotide sequence encoding a FMN1 polypeptide, wherein the nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:38.

[00376] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more flavin mononucleotide (FMN) synthetase polypeptides, the one or more flavin mononucleotide (FMN) synthetase polypeptides are encoded by a nucleotide sequence encoding a FMN1 polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:38.

Glycosidase Polypeptides

[00377] Modified host cells of the disclosure may comprise one or more modifications to modulate the expression of one or more glycosidase polypeptides. The one or more modifications to modulate the expression of one or more glycosidase polypeptides may include introducing into a host cell one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosidase polypeptides and/or deletion or downregulation of one or more genes encoding one or more glycosidase polypeptides in a host cell. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosidase polypeptides, resulting in expression or overexpression of the one or more glycosidase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more glycosidase polypeptides, reducing or eliminating the expression of the one or more glycosidase polypeptides. Glycosidase polypeptides (also may be referred to as glycoside hydrolase polypeptides or glycosyl hydrolase polypeptides) catalyze the hydrolysis of glycosidic bonds in carbohydrates.

[00378] Exemplary glycosidase polypeptides disclosed herein may also include a full- length glycosidase polypeptide, a fragment of a glycosidase polypeptide, a variant of a glycosidase polypeptide, a truncated glycosidase polypeptide, or a fusion polypeptide that has at least one activity of a glycosidase polypeptide.

[00379] Modified host cells of the disclosure may comprise one or more modifications to modulate the expression of one or more glycosidase polypeptides including, but not limited to, glucosidase polypeptides (e.g., a ROT2 polypeptide or a CWH41 polypeptide) and mannosidase polypeptides (e.g., a MNS1 polypeptide or a HTMl polypeptide).

[00380] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosidase polypeptides, the one or more glycosidase polypeptides are selected from the group consisting of: a ROT2 polypeptide, a MNS1 polypeptide, a HTM1 polypeptide, and a CWH41 polypeptide.

[00381] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosidase polypeptides, the one or more glycosidase polypeptides are selected from the group consisting of glycosidase polypeptides comprising the amino acid sequences set forth in SEQ ID NO:203 (a ROT2 polypeptide), SEQ ID NO:207 (a MNS1 polypeptide), SEQ ID NO:227 (a HTMl polypeptide), and SEQ ID NO:43 (a CWH41 polypeptide).

[00382] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosidase polypeptides, the one or more glycosidase polypeptides are selected from the group consisting of glycosidase polypeptides comprising the amino acid sequences set forth in SEQ ID NO:203 (a ROT2 polypeptide), SEQ ID NO:207 (a MNS1 polypeptide), SEQ ID NO:227 (a HTMl polypeptide), and SEQ ID NO:43 (a CWH41 polypeptide), or a conservatively substituted amino acid sequence of any of the foregoing.

[00383] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosidase polypeptides, the one or more glycosidase polypeptides are selected from the group consisting of glycosidase polypeptides comprising amino acid sequences having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to: SEQ ID NO:203 (a ROT2 polypeptide), SEQ ID NO:207 (a MNS1 polypeptide), SEQ ID NO:227 (a HTM1 polypeptide), and SEQ ID NO:43 (a CWH41 polypeptide).

[00384] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more glycosidase polypeptides, the one or more glycosidase polypeptides are selected from the group consisting of a ROT2 polypeptide, a HTM1 polypeptide, a MNS1 polypeptide, and a CWH41 polypeptide. [00385] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more glycosidase polypeptides, the one or more glycosidase polypeptides are selected from the group consisting of glycosidase polypeptides comprising the amino acid sequences set forth in SEQ ID NO: 203 (a ROT2 polypeptide), SEQ ID NO: 207 (a MNS1 polypeptide), SEQ ID NO:227 (a HTM1 polypeptide), and SEQ ID NO:43 (a CWH41 polypeptide).

[00386] In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a glycosidase polypeptide. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosidase polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding two or more glycosidase polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding three or more glycosidase polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding four or more glycosidase polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding five or more glycosidase polypeptides.

[00387] In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more glycosidase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of two or more genes encoding two or more glycosidase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of three or more genes encoding three or more glycosidase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of four or more genes encoding four or more glycosidase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of five or more genes encoding five or more glycosidase polypeptides.

[00388] In some embodiments, the nucleotide sequences encoding the one or more glycosidase polypeptides are codon-optimized. [00389] In some embodiments, one or more glycosidase polypeptides are

overexpressed in the modified host cell. Overexpression may be achieved by increasing the copy number of the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosidase polypeptides, e.g., through use of a high copy number expression vector (e.g., a plasmid that exists at 10-40 copies or about 100 copies per cell) and/or by operably linking the nucleotide sequences encoding the one or more glycosidase polypeptides to a strong promoter. In some embodiments, the modified host cell has one copy of a heterologous nucleic acid comprising a nucleotide sequence encoding a glycosidase polypeptide. In some embodiments, the modified host cell has two copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a glycosidase polypeptide. In some embodiments, the modified host cell has three copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a glycosidase polypeptide. In some embodiments, the modified host cell has four copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a glycosidase polypeptide. In some

embodiments, the modified host cell has five copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a glycosidase polypeptide. In some

embodiments, the modified host cell has five or more copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a glycosidase polypeptide. Increased copy number of the heterologous nucleic acid and/or codon optimization of the nucleotide sequence may result in an increase in the desired polypeptide activity in the modified host cell.

[00390] Exemplary heterologous nucleic acids disclosed herein may include nucleic acids comprising a nucleotide sequence that encodes a glycosidase polypeptide, such as, a full-length glycosidase polypeptide, a fragment of a glycosidase polypeptide, a variant of a glycosidase polypeptide, a truncated glycosidase polypeptide, or a fusion polypeptide that has at least one activity of a glycosidase polypeptide. In some embodiments, the nucleotide sequence is codon-optimized.

[00391] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosidase polypeptides, the nucleotide sequences encoding the one or more glycosidase polypeptides are selected from the group consisting of nucleotide sequences set forth in SEQ ID NO:202 (encodes a ROT2 polypeptide), SEQ ID NO:206 (encodes a MNS1 polypeptide), SEQ ID NO:226 (encodes a HTM1 polypeptide), and SEQ ID NO:42 (encodes a CWH41 polypeptide). [00392] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosidase polypeptides, the nucleotide sequences encoding the one or more glycosidase polypeptides are selected from the group consisting of nucleotide sequences set forth in SEQ ID NO:202 (encodes a ROT2 polypeptide), SEQ ID NO:206 (encodes a MNS1 polypeptide), SEQ ID NO:226 (encodes a HTM1 polypeptide), and SEQ ID NO:42 (encodes a CWH41 polypeptide), or a codon degenerate nucleotide sequence of any of the foregoing.

[00393] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosidase polypeptides, the nucleotide sequences encoding the one or more glycosidase polypeptides are selected from the group consisting of nucleotide sequences having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to: SEQ ID NO:202 (encodes a ROT2 polypeptide), SEQ ID NO:206 (encodes a MNS1 polypeptide), SEQ ID NO:226 (encodes a HTM1 polypeptide), and SEQ ID NO:42 (encodes a CWH41 polypeptide).

[00394] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more glycosidase polypeptides, the one or more glycosidase polypeptides are encoded by nucleotide sequences selected from the group consisting of nucleotide sequences set forth in SEQ ID NO:202 (encodes a ROT2 polypeptide), SEQ ID NO:206 (encodes a MNS1 polypeptide), SEQ ID NO:226 (encodes a HTM1 polypeptide), and SEQ ID NO:42 (encodes a CWH41 polypeptide).

[00395] In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more of the following genes: a ROT2 gene or a MNS1 gene. In some embodiments, the modified host cells of the disclosure comprise a deletion of one or more of the following genes: a ROT2 gene or a MNS1 gene. In some embodiments, the modified host cells of the disclosure comprise a downregulation of one or more of the following genes: a ROT2 gene or a MNS1 gene. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of a ROT2 gene. In some embodiments, the modified host cells of the disclosure comprise a deletion of a ROT2 gene. In some embodiments, the modified host cells of the disclosure comprise a downregulation of a ROT2 gene. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of a MNS1 gene. In some embodiments, the modified host cells of the disclosure comprise a deletion of a MNS1 gene. In some embodiments, the modified host cells of the disclosure comprise a downregulation of a MNS1 gene.

Glycosyltransj erase Polypeptides

[00396] Modified host cells of the disclosure may comprise one or more modifications to modulate the expression of one or more glycosyltransferase polypeptides. The one or more modifications to modulate the expression of one or more glycosyltransferase polypeptides may include introducing into a host cell one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosyltransferase polypeptides and/or deletion or downregulation of one or more genes encoding one or more

glycosyltransferase polypeptides in a host cell. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosyltransferase polypeptides, resulting in expression or overexpression of the one or more glycosyltransferase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or

downregulation of one or more genes encoding one or more glycosyltransferase

polypeptides, reducing or eliminating the expression of the one or more glycosyltransferase polypeptides. Glycosyltransferase polypeptides (also may be known as

oligosaccharyltransferase polypeptides) catalyze the formation of glycosidic linkages.

[00397] Exemplary glycosyltransferase polypeptides disclosed herein may also include a full-length glycosyltransferase polypeptide, a fragment of a glycosyltransferase polypeptide, a variant of a glycosyltransferase polypeptide, a truncated glycosyltransferase polypeptide, or a fusion polypeptide that has at least one activity of a glycosyltransferase polypeptide.

[00398] Modified host cells of the disclosure may comprise one or more modifications to modulate the expression of one or more glycosyltransferase polypeptides including, but not limited to, mannosyltransferase polypeptides (e.g., an ALG12 polypeptide).

[00399] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosyltransferase polypeptides, the one or more glycosyltransferase polypeptides is an ALG12 polypeptide.

[00400] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosyltransferase polypeptides, the one or more glycosyltransferase polypeptides is an ALG12 polypeptide comprising the amino acid sequence set forth in SEQ ID NO:205.

[00401] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosyltransferase polypeptides, the one or more glycosyltransferase polypeptides is an ALG12 polypeptide comprising the amino acid sequence set forth in SEQ ID NO:205, or a conservatively substituted amino acid sequence thereof.

[00402] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosyltransferase polypeptides, the one or more glycosyltransferase polypeptides is an ALG12 polypeptide comprising an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:205.

[00403] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more glycosyltransferase polypeptides, the one or more glycosyltransferase polypeptides is an ALG12 polypeptide.

[00404] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more glycosyltransferase polypeptides, the one or more glycosyltransferase polypeptides is an ALG12 polypeptide comprising the amino acid sequence set forth in SEQ ID NO:205.

[00405] In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a glycosyltransferase polypeptide. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosyltransferase polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding two or more glycosyltransferase polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding three or more glycosyltransferase polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding four or more glycosyltransferase polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding five or more glycosyltransferase polypeptides.

[00406] In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more glycosyltransferase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of two or more genes encoding two or more glycosyltransferase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of three or more genes encoding three or more

glycosyltransferase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of four or more genes encoding four or more glycosyltransferase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of five or more genes encoding five or more glycosyltransferase polypeptides.

[00407] In some embodiments, the nucleotide sequences encoding the one or more glycosyltransferase polypeptides are codon-optimized.

[00408] In some embodiments, one or more glycosyltransferase polypeptides are overexpressed in the modified host cell. Overexpression may be achieved by increasing the copy number of the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosyltransferase polypeptides, e.g., through use of a high copy number expression vector (e g., a plasmid that exists at 10-40 copies or about 100 copies per cell) and/or by operably linking the nucleotide sequences encoding the one or more glycosyltransferase polypeptides to a strong promoter. In some embodiments, the modified host cell has one copy of a heterologous nucleic acid comprising a nucleotide sequence encoding a glycosyltransferase polypeptide. In some embodiments, the modified host cell has two copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a glycosyltransferase polypeptide. In some embodiments, the modified host cell has three copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a glycosyltransferase polypeptide. In some embodiments, the modified host cell has four copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a glycosyltransferase polypeptide. In some embodiments, the modified host cell has five copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a glycosyltransferase polypeptide. In some embodiments, the modified host cell has five or more copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a glycosyltransferase polypeptide. Increased copy number of the heterologous nucleic acid and/or codon optimization of the nucleotide sequence may result in an increase in the desired polypeptide activity in the modified host cell.

[00409] Exemplary heterologous nucleic acids disclosed herein may include nucleic acids comprising a nucleotide sequence that encodes a glycosyltransferase polypeptide, such as, a full-length glycosyltransferase polypeptide, a fragment of a glycosyltransferase polypeptide, a variant of a glycosyltransferase polypeptide, a truncated glycosyltransferase polypeptide, or a fusion polypeptide that has at least one activity of a glycosyltransferase polypeptide. In some embodiments, the nucleotide sequence is codon-optimized.

[00410] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosyltransferase polypeptides, the nucleotide sequences encoding the one or more glycosyltransferase polypeptides is a nucleotide sequence encoding an ALG12 polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:204.

[00411] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosyltransferase polypeptides, the nucleotide sequences encoding the one or more glycosyltransferase polypeptides is a nucleotide sequence encoding an ALG12 polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:204, or a codon degenerate nucleotide sequence thereof.

[00412] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosyltransferase polypeptides, the nucleotide sequences encoding the one or more glycosyltransferase polypeptides is a nucleotide sequence encoding an ALG12 polypeptide, wherein the nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:204. [00413] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more glycosyltransferase polypeptides, the one or more glycosyltransferase polypeptides are encoded by a nucleotide sequence encoding an ALG12 polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:204.

[00414] In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of an ALG12 gene. In some embodiments, the modified host cells of the disclosure comprise a deletion of an ALG12 gene. In some embodiments, the modified host cells of the disclosure comprise a downregulation of an ALG12 gene.

Protein Disulfide Isomerase Polypeptides

[00415] Modified host cells of the disclosure may comprise one or more modifications to modulate the expression of one or more protein disulfide isomerase polypeptides. The one or more modifications to modulate the expression of one or more protein disulfide isomerase polypeptides may include introducing into a host cell one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides and/or deletion or downregulation of one or more genes encoding one or more protein disulfide isomerase polypeptides in a host cell. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, resulting in expression or overexpression of the one or more protein disulfide isomerase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more protein disulfide isomerase polypeptides, reducing or eliminating the expression of the one or more protein disulfide isomerase polypeptides. Protein disulfide isomerase polypeptides may catalyze the formation and breakage of disulfide bonds between cysteine residues within polypeptides as they fold.

[00416] Exemplary protein disulfide isomerase polypeptides disclosed herein may also include a full-length protein disulfide isomerase polypeptide, a fragment of a protein disulfide isomerase polypeptide, a variant of a protein disulfide isomerase polypeptide, a truncated protein disulfide isomerase polypeptide, or a fusion polypeptide that has at least one activity of a protein disulfide isomerase polypeptide.

[00417] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, the one or more protein disulfide isomerase polypeptides is a PDI1 polypeptide.

[00418] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, the one or more protein disulfide isomerase polypeptides is a PDI1 polypeptide comprising the amino acid sequence set forth in SEQ ID NO:55.

[00419] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, the one or more protein disulfide isomerase polypeptides is a PDI1 polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 55, or a conservatively substituted amino acid sequence thereof.

[00420] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, the one or more protein disulfide isomerase polypeptides is a PDI1 polypeptide comprising an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:55.

[00421] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more protein disulfide isomerase polypeptides, the one or more protein disulfide isomerase polypeptides is a PDI1 polypeptide.

[00422] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more protein disulfide isomerase polypeptides, the one or more protein disulfide isomerase polypeptides is a PDI1 polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 55.

[00423] In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a protein disulfide isomerase polypeptide. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding two or more protein disulfide isomerase polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding three or more protein disulfide isomerase polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding four or more protein disulfide isomerase polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding five or more protein disulfide isomerase polypeptides.

[00424] In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more protein disulfide isomerase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of two or more genes encoding two or more protein disulfide isomerase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of three or more genes encoding three or more protein disulfide isomerase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of four or more genes encoding four or more protein disulfide isomerase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of five or more genes encoding five or more protein disulfide isomerase polypeptides.

[00425] In some embodiments, the nucleotide sequences encoding the one or more protein disulfide isomerase polypeptides are codon-optimized.

[00426] In some embodiments, one or more protein disulfide isomerase polypeptides are overexpressed in the modified host cell. Overexpression may be achieved by increasing the copy number of the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, e.g., through use of a high copy number expression vector (e g., a plasmid that exists at 10-40 copies or about 100 copies per cell) and/or by operably linking the nucleotide sequences encoding the one or more protein disulfide isomerase polypeptides to a strong promoter. In some embodiments, the modified host cell has one copy of a heterologous nucleic acid comprising a nucleotide sequence encoding a protein disulfide isomerase polypeptide. In some embodiments, the modified host cell has two copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a protein disulfide isomerase polypeptide. In some embodiments, the modified host cell has three copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a protein disulfide isomerase polypeptide. In some embodiments, the modified host cell has four copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a protein disulfide isomerase polypeptide. In some embodiments, the modified host cell has five copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a protein disulfide isomerase polypeptide. In some embodiments, the modified host cell has five or more copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a protein disulfide isomerase polypeptide. Increased copy number of the heterologous nucleic acid and/or codon optimization of the nucleotide sequence may result in an increase in the desired polypeptide activity in the modified host cell.

[00427] Exemplary heterologous nucleic acids disclosed herein may include nucleic acids comprising a nucleotide sequence that encodes a protein disulfide isomerase polypeptide, such as, a full-length protein disulfide isomerase polypeptide, a fragment of a protein disulfide isomerase polypeptide, a variant of a protein disulfide isomerase polypeptide, a truncated protein disulfide isomerase polypeptide, or a fusion polypeptide that has at least one activity of a protein disulfide isomerase polypeptide. In some embodiments, the nucleotide sequence is codon-optimized.

[00428] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, the nucleotide sequences encoding the one or more protein disulfide isomerase polypeptides is a nucleotide sequence encoding a PDI1 polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO: 54.

[00429] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, the nucleotide sequences encoding the one or more protein disulfide isomerase polypeptides is a nucleotide sequence encoding a PDI1 polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO: 54, or a codon degenerate nucleotide sequence thereof.

[00430] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, the nucleotide sequences encoding the one or more protein disulfide isomerase polypeptides is a nucleotide sequence encoding a PDI1 polypeptide, wherein the nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:54.

[00431] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more protein disulfide isomerase polypeptides, the one or more protein disulfide isomerase polypeptides are encoded by a nucleotide sequence encoding a PDI1 polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:54.

Thiol Oxidase Polypeptides

[00432] Modified host cells of the disclosure may comprise one or more modifications to modulate the expression of one or more thiol oxidase polypeptides. The one or more modifications to modulate the expression of one or more thiol oxidase polypeptides may include introducing into a host cell one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides and/or deletion or downregulation of one or more genes encoding one or more thiol oxidase polypeptides in a host cell. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, resulting in expression or overexpression of the one or more thiol oxidase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more thiol oxidase polypeptides, reducing or eliminating the expression of the one or more thiol oxidase polypeptides. Thiol oxidase polypeptides transfer electrons from reduced protein disulfide isomerase polypeptides to a terminal acceptor such as oxygen.

[00433] Exemplary thiol oxidase polypeptides disclosed herein may also include a full-length thiol oxidase polypeptide, a fragment of a thiol oxidase polypeptide, a variant of a thiol oxidase polypeptide, a truncated thiol oxidase polypeptide, or a fusion polypeptide that has at least one activity of a thiol oxidase polypeptide.

[00434] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, the one or more thiol oxidase polypeptides are selected from the group consisting of: an EROl polypeptide and an ERV2 polypeptide. The EROl and ERV2 polypeptides may serve as partners to the PDI1 polypeptide, a protein disulfide isomerase polypeptide.

[00435] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, the one or more thiol oxidase polypeptides are selected from the group consisting of thiol oxidase polypeptides comprising the amino acid sequences set forth in SEQ ID NO:5 l (an EROl polypeptide) and SEQ ID NO:53 (an ERV2 polypeptide).

[00436] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, the one or more thiol oxidase polypeptides are selected from the group consisting of thiol oxidase polypeptides comprising the amino acid sequences set forth in SEQ ID NO:5 l (an EROl polypeptide) and SEQ ID NO:53 (an ERV2 polypeptide), or a conservatively substituted amino acid sequence of any of the foregoing.

[00437] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, the one or more thiol oxidase polypeptides are selected from the group consisting of thiol oxidase polypeptides comprising amino acid sequences having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO: 51 (an EROl polypeptide) and SEQ ID NO: 53 (an ERV2 polypeptide).

[00438] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more thiol oxidase polypeptides, the one or more thiol oxidase polypeptides are selected from the group consisting of an EROl polypeptide and an ERV2 polypeptide.

[00439] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more thiol oxidase polypeptides, the one or more thiol oxidase polypeptides are selected from the group consisting of thiol oxidase polypeptides comprising the amino acid sequences set forth in SEQ ID NO:51 (an EROl polypeptide) and SEQ ID NO:53 (an ERV2 polypeptide). [00440] In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding a thiol oxidase polypeptide. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding two or more thiol oxidase polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding three or more thiol oxidase polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding four or more thiol oxidase polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding five or more thiol oxidase polypeptides.

[00441] In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more thiol oxidase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of two or more genes encoding two or more thiol oxidase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of three or more genes encoding three or more thiol oxidase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of four or more genes encoding four or more thiol oxidase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of five or more genes encoding five or more thiol oxidase polypeptides.

[00442] In some embodiments, the nucleotide sequences encoding the one or more thiol oxidase polypeptides are codon-optimized.

[00443] In some embodiments, one or more thiol oxidase polypeptides are overexpressed in the modified host cell. Overexpression may be achieved by increasing the copy number of the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, e.g., through use of a high copy number expression vector (e.g., a plasmid that exists at 10-40 copies or about 100 copies per cell) and/or by operably linking the nucleotide sequences encoding the one or more thiol oxidase polypeptides to a strong promoter. In some embodiments, the modified host cell has one copy of a heterologous nucleic acid comprising a nucleotide sequence encoding a thiol oxidase polypeptide. In some embodiments, the modified host cell has two copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a thiol oxidase polypeptide. In some embodiments, the modified host cell has three copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a thiol oxidase polypeptide. In some embodiments, the modified host cell has four copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a thiol oxidase polypeptide. In some embodiments, the modified host cell has five copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a thiol oxidase polypeptide. In some

embodiments, the modified host cell has five or more copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a thiol oxidase polypeptide. Increased copy number of the heterologous nucleic acid and/or codon optimization of the nucleotide sequence may result in an increase in the desired polypeptide activity in the modified host cell.

[00444] Exemplary heterologous nucleic acids disclosed herein may include nucleic acids comprising a nucleotide sequence that encodes a thiol oxidase polypeptide, such as, a full-length thiol oxidase polypeptide, a fragment of a thiol oxidase polypeptide, a variant of a thiol oxidase polypeptide, a truncated thiol oxidase polypeptide, or a fusion polypeptide that has at least one activity of a thiol oxidase polypeptide. In some embodiments, the nucleotide sequence is codon-optimized.

[00445] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, the nucleotide sequences encoding the one or more thiol oxidase polypeptides are selected from the group consisting of nucleotide sequences set forth in SEQ ID NO:50 (encodes an EROl polypeptide) and SEQ ID NO:52 (encodes an ERV2 polypeptide).

[00446] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, the nucleotide sequences encoding the one or more thiol oxidase polypeptides are selected from the group consisting of nucleotide sequences set forth in SEQ ID NO:50 (encodes an EROl polypeptide) and SEQ ID NO:52 (encodes an ERV2 polypeptide), or a codon degenerate nucleotide sequence of any of the foregoing.

[00447] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, the nucleotide sequences encoding the one or more thiol oxidase polypeptides are selected from the group consisting of nucleotide sequences having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO: 50 (encodes an EROl polypeptide) and SEQ ID NO: 52 (encodes an ERV2 polypeptide).

[00448] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more thiol oxidase polypeptides, the one or more thiol oxidase polypeptides are encoded by nucleotide sequences selected from the group consisting of nucleotide sequences set forth in SEQ ID NO: 50 (encodes an EROl polypeptide) and SEQ ID NO: 52 (encodes an ERV2 polypeptide).

Fatty Acid Desaturase Polypeptides

[00449] Modified host cells of the disclosure may comprise one or more modifications to modulate the expression of one or more fatty acid desaturase polypeptides. The one or more modifications to modulate the expression of one or more fatty acid desaturase polypeptides may include introducing into a host cell one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more fatty acid desaturase polypeptides and/or deletion or downregulation of one or more genes encoding one or more fatty acid desaturase polypeptides in a host cell. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more fatty acid desaturase polypeptides, resulting in expression or overexpression of the one or more fatty acid desaturase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or

downregulation of one or more genes encoding one or more fatty acid desaturase

polypeptides, reducing or eliminating the expression of the one or more fatty acid desaturase polypeptides. A fatty acid desaturase polypeptide may remove two hydrogen atoms from a fatty acid, forming a carbon-carbon double bond.

[00450] Exemplary fatty acid desaturase polypeptides disclosed herein may also include a full-length fatty acid desaturase polypeptide, a fragment of a fatty acid desaturase polypeptide, a variant of a fatty acid desaturase polypeptide, a truncated fatty acid desaturase polypeptide, or a fusion polypeptide that has at least one activity of a fatty acid desaturase polypeptide. [00451] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more fatty acid desaturase polypeptides, the one or more fatty acid desaturase polypeptides is an OLE1 polypeptide.

[00452] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more fatty acid desaturase polypeptides, the one or more fatty acid desaturase polypeptides is an OLE1 polypeptide comprising the amino acid sequence set forth in SEQ ID NO:45.

[00453] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more fatty acid desaturase polypeptides, the one or more fatty acid desaturase polypeptides is an OLE1 polypeptide comprising the amino acid sequence set forth in SEQ ID NO:45, or a conservatively substituted amino acid sequence thereof.

[00454] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more fatty acid desaturase polypeptides, the one or more fatty acid desaturase polypeptides is an OLE1 polypeptide comprising an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99 8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:45.

[00455] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more fatty acid desaturase polypeptides, the one or more fatty acid desaturase polypeptides is an OLE1 polypeptide.

[00456] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more fatty acid desaturase polypeptides, the one or more fatty acid desaturase polypeptides is an OLE1 polypeptide comprising the amino acid sequence set forth in SEQ ID NO:45.

[00457] In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a fatty acid desaturase polypeptide. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more fatty acid desaturase polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding two or more fatty acid desaturase polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding three or more fatty acid desaturase polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding four or more fatty acid desaturase polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding five or more fatty acid desaturase polypeptides.

[00458] In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more fatty acid desaturase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of two or more genes encoding two or more fatty acid desaturase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of three or more genes encoding three or more fatty acid desaturase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of four or more genes encoding four or more fatty acid desaturase polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of five or more genes encoding five or more fatty acid desaturase polypeptides.

[00459] In some embodiments, the nucleotide sequences encoding the one or more fatty acid desaturase polypeptides are codon-optimized.

[00460] In some embodiments, one or more fatty acid desaturase polypeptides are overexpressed in the modified host cell. Overexpression may be achieved by increasing the copy number of the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more fatty acid desaturase polypeptides, e.g., through use of a high copy number expression vector (e g., a plasmid that exists at 10-40 copies or about 100 copies per cell) and/or by operably linking the nucleotide sequences encoding the one or more fatty acid desaturase polypeptides to a strong promoter. In some embodiments, the modified host cell has one copy of a heterologous nucleic acid comprising a nucleotide sequence encoding a fatty acid desaturase polypeptide. In some embodiments, the modified host cell has two copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a fatty acid desaturase polypeptide. In some embodiments, the modified host cell has three copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a fatty acid desaturase polypeptide. In some embodiments, the modified host cell has four copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a fatty acid desaturase polypeptide. In some embodiments, the modified host cell has five copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a fatty acid desaturase polypeptide. In some embodiments, the modified host cell has five or more copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a fatty acid desaturase polypeptide. Increased copy number of the heterologous nucleic acid and/or codon optimization of the nucleotide sequence may result in an increase in the desired polypeptide activity in the modified host cell.

[00461] Exemplary heterologous nucleic acids disclosed herein may include nucleic acids comprising a nucleotide sequence that encodes a fatty acid desaturase polypeptide, such as, a full-length fatty acid desaturase polypeptide, a fragment of a fatty acid desaturase polypeptide, a variant of a fatty acid desaturase polypeptide, a truncated fatty acid desaturase polypeptide, or a fusion polypeptide that has at least one activity of a fatty acid desaturase polypeptide. In some embodiments, the nucleotide sequence is codon-optimized.

[00462] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more fatty acid desaturase polypeptides, the nucleotide sequences encoding the one or more fatty acid desaturase polypeptides is a nucleotide sequence encoding an OLE1 polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:44.

[00463] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more fatty acid desaturase polypeptides, the nucleotide sequences encoding the one or more fatty acid desaturase polypeptides is a nucleotide sequence encoding an OLE1 polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:44, or a codon degenerate nucleotide sequence thereof.

[00464] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more fatty acid desaturase polypeptides, the nucleotide sequences encoding the one or more fatty acid desaturase polypeptides is a nucleotide sequence encoding an OLE1 polypeptide, wherein the nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:44.

[00465] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more fatty acid desaturase polypeptides, the one or more fatty acid desaturase polypeptides are encoded by a nucleotide sequence encoding an OLE1 polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:44.

Protein Transport and Trafficking Polypeptides

[00466] Modified host cells of the disclosure may comprise one or more modifications to modulate the expression of one or more protein transport and trafficking polypeptides.

The one or more modifications to modulate the expression of one or more protein transport and trafficking polypeptides may include introducing into a host cell one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein transport and trafficking polypeptides and/or deletion or downregulation of one or more genes encoding one or more protein transport and trafficking polypeptides in a host cell. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein transport and trafficking polypeptides, resulting in expression or overexpression of the one or more protein transport and trafficking polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more protein transport and trafficking polypeptides, reducing or eliminating the expression of the one or more protein transport and trafficking polypeptides.

[00467] Exemplary protein transport and trafficking polypeptides disclosed herein may also include a full-length protein transport and trafficking polypeptide, a fragment of a protein transport and trafficking polypeptide, a variant of a protein transport and trafficking polypeptide, a truncated protein transport and trafficking polypeptide, or a fusion polypeptide that has at least one activity of a protein transport and trafficking polypeptide.

[00468] Modified host cells of the disclosure may comprise one or more modifications to modulate the expression of one or more protein transport and trafficking polypeptides including, but not limited to, polypeptides involved in ER to Golgi transit polypeptides (e.g., a SEC7 polypeptide, a BFR2 polypeptide, a COPII complex polypeptide (e.g., a SEC23 polypeptide, a SEC24 polypeptide, a SEC13 polypeptide, a SEC31 polypeptide, a SARI polypeptide, a SEC 12 polypeptide, a SEC4 polypeptide, a SEC 16 polypeptide, or an ERV29 polypeptide) and a SNARES complex polypeptide (e.g., a SEC5 polypeptide, a SEC22 polypeptide, a BOSl polypeptide, a BETl polypeptide, a BElGl polypeptide, a GRH1 polypeptide, a USOl polypeptide, a SEC17 polypeptide, a SEC18 polypeptide, a SSOl polypeptide, a SEC9 polypeptide, or a SNC2 polypeptide)).

[00469] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein transport and trafficking polypeptides, the one or more protein transport and trafficking polypeptides are selected from the group consisting of: a SEC23 polypeptide, a SEC24 polypeptide, a SEC13 polypeptide, a BFR2 polypeptide, a SEC31 polypeptide, a SAR1 polypeptide, a SEC 12 polypeptide, a SEC4 polypeptide, a SEC 16 polypeptide, an ERV29 polypeptide, a SEC7 polypeptide, a SEC5 polypeptide, a SEC22 polypeptide, a BOS1 polypeptide, a BETl polypeptide, a BElGl polypeptide, a GRH1 polypeptide, a USOl polypeptide, a SEC 17 polypeptide, a SEC 18 polypeptide, a SSOl polypeptide, a SEC9 polypeptide, a SNC2 polypeptide, and a LDB17 polypeptide.

[00470] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein transport and trafficking polypeptides, the one or more protein transport and trafficking polypeptides are selected from the group consisting of protein transport and trafficking polypeptides comprising the amino acid sequences set forth in SEQ ID NO:59 (a SEC23 polypeptide), SEQ ID NO:6l (a SEC24 polypeptide), SEQ ID NO:63 (a SEC 13 polypeptide), SEQ ID NO:21 (a BFR2 polypeptide), SEQ ID NO: 155 (a SEC31

polypeptide), SEQ ID NO: l57 (a SAR1 polypeptide), SEQ ID NOT59 (a SEC 12 polypeptide), SEQ ID NO: 161 (a SEC4 polypeptide), SEQ ID NO: 163 (a SEC 16 polypeptide), SEQ ID NO: 165 (an ERV29 polypeptide), SEQ ID NO: 167 (a SEC7 polypeptide), SEQ ID NO: 169 (a SEC 5 polypeptide), SEQ ID NO: 171 (a SEC22 polypeptide), SEQ ID NO: 173 (a BOSl polypeptide), SEQ ID NO: 175 (a BETl

polypeptide), SEQ ID NO: 177 (a BUG1 polypeptide), SEQ ID NO: 179 (a GRHl polypeptide), SEQ ID NO: 181 (a USOl polypeptide), SEQ ID NO: l83 (a SEC 17 polypeptide), SEQ ID NO: l85 (a SEC 18 polypeptide), SEQ ID NO: l87 (a SSOl polypeptide), SEQ ID NO: 189 (a SEC 9 polypeptide), SEQ ID NO: 191 (a SNC2

polypeptide), and SEQ ID NO: 193 (a LDBl7 polypeptide). [00471] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein transport and trafficking polypeptides, the one or more protein transport and trafficking polypeptides are selected from the group consisting of protein transport and trafficking polypeptides comprising the amino acid sequences set forth in SEQ ID NO:59 (a SEC23 polypeptide), SEQ ID NO:6l (a SEC24 polypeptide), SEQ ID NO:63 (a SEC 13 polypeptide), SEQ ID NO:21 (a BFR2 polypeptide), SEQ ID NO: 155 (a SEC31

polypeptide), SEQ ID NO: l57 (a SAR1 polypeptide), SEQ ID NO: l59 (a SEC 12 polypeptide), SEQ ID NO: 161 (a SEC4 polypeptide), SEQ ID NO: 163 (a SEC 16 polypeptide), SEQ ID NO: 165 (an ERV29 polypeptide), SEQ ID NO: 167 (a SEC7 polypeptide), SEQ ID NO: 169 (a SEC5 polypeptide), SEQ ID NO: 171 (a SEC22 polypeptide), SEQ ID NO: 173 (a BOSl polypeptide), SEQ ID NO: 175 (a BET1

polypeptide), SEQ ID NO: 177 (a BUG1 polypeptide), SEQ ID NO: 179 (a GRH1 polypeptide), SEQ ID NO: 181 (a USOl polypeptide), SEQ ID NO: l83 (a SEC17 polypeptide), SEQ ID NO: 185 (a SEC 18 polypeptide), SEQ ID NO: 187 (a SSOl polypeptide), SEQ ID NO: 189 (a SEC 9 polypeptide), SEQ ID NO: 191 (a SNC2

polypeptide), and SEQ ID NO: 193 (a LDB 17 polypeptide), or a conservatively substituted amino acid sequence of any of the foregoing.

[00472] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein transport and trafficking polypeptides, the one or more protein transport and trafficking polypeptides are selected from the group consisting of protein transport and trafficking polypeptides comprising amino acid sequences having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:59 (a SEC23 polypeptide), SEQ ID NO:6l (a SEC24 polypeptide), SEQ ID NO:63 (a SEC 13 polypeptide), SEQ ID NO:21 (a BFR2 polypeptide), SEQ ID NO: 155 (a SEC31 polypeptide), SEQ ID NO: l57 (a SAR1 polypeptide), SEQ ID NO: l59 (a SEC 12 polypeptide), SEQ ID NO: 161 (a SEC4 polypeptide), SEQ ID NO: 163 (a SEC 16 polypeptide), SEQ ID NO: 165 (an ERV29 polypeptide), SEQ ID NO: 167 (a SEC7 polypeptide), SEQ ID NO: 169 (a SEC 5 polypeptide), SEQ ID NO: 171 (a SEC22 polypeptide), SEQ ID NO: 173 (a BOSl polypeptide), SEQ ID NO: 175 (a BETl polypeptide), SEQ ID NO: 177 (a BUG1 polypeptide), SEQ ID NO: 179 (a GRH1 polypeptide), SEQ ID NO: 181 (a USOl polypeptide), SEQ ID NO: l83 (a SEC 17 polypeptide), SEQ ID NO: l85 (a SEC 18 polypeptide), SEQ ID NO: l87 (a SSOl polypeptide), SEQ ID NO: 189 (a SEC 9 polypeptide), SEQ ID NO: 191 (a SNC2 polypeptide), and SEQ ID NO: 193 (a LDB17 polypeptide).

[00473] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more protein transport and trafficking polypeptides, the one or more protein transport and trafficking polypeptides are selected from the group consisting of a SEC23 polypeptide, a SEC24 polypeptide, a SEC 13 polypeptide, a BFR2 polypeptide, a SEC31 polypeptide, a SARI polypeptide, a SEC 12 polypeptide, a SEC4 polypeptide, a SEC 16 polypeptide, an ERV29 polypeptide, a SEC7 polypeptide, a SEC5 polypeptide, a SEC22 polypeptide, a BOSl polypeptide, a BETl polypeptide, a BUGl polypeptide, a GRH1 polypeptide, a USOl polypeptide, a SEC 17 polypeptide, a SEC 18 polypeptide, a SSOl polypeptide, a SEC9 polypeptide, a SNC2 polypeptide, and a LDB17 polypeptide.

[00474] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more protein transport and trafficking polypeptides, the one or more protein transport and trafficking polypeptides are selected from the group consisting of protein transport and trafficking polypeptides comprising the amino acid sequences set forth in SEQ ID NO:59 (a SEC23 polypeptide), SEQ ID NO:61 (a SEC24 polypeptide), SEQ ID NO:63 (a SEC 13 polypeptide), SEQ ID NO:21 (a BFR2 polypeptide), SEQ ID NO: 155 (a SEC31 polypeptide), SEQ ID NO: 157 (a SARI polypeptide), SEQ ID NO: 159 (a SEC 12 polypeptide), SEQ ID NO: 161 (a SEC4 polypeptide), SEQ ID NO: 163 (a SEC 16 polypeptide), SEQ ID NO: 165 (an ERV29 polypeptide), SEQ ID NO: 167 (a SEC7 polypeptide), SEQ ID NO: 169 (a SEC 5 polypeptide), SEQ ID NO: 171 (a SEC22 polypeptide), SEQ ID NO: 173 (a BOSl polypeptide), SEQ ID NO: 175 (a BETl polypeptide), SEQ ID NO: 177 (a BUG1 polypeptide), SEQ ID NO: 179 (a GRH1 polypeptide), SEQ ID NO: 181 (a USOl polypeptide), SEQ ID NO: 183 (a SEC 17 polypeptide), SEQ ID NO: 185 (a SEC 18 polypeptide), SEQ ID NO: 187 (a SSOl polypeptide), SEQ ID NO: 189 (a SEC 9 polypeptide), SEQ ID NO: 191 (a SNC2 polypeptide), and SEQ ID NO: 193 (a LDB17 polypeptide). [00475] In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a protein transport and trafficking polypeptide. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein transport and trafficking polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding two or more protein transport and trafficking polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding three or more protein transport and trafficking polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding four or more protein transport and trafficking polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding five or more protein transport and trafficking polypeptides.

[00476] In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more protein transport and trafficking polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of two or more genes encoding two or more protein transport and trafficking polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of three or more genes encoding three or more protein transport and trafficking polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of four or more genes encoding four or more protein transport and trafficking polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of five or more genes encoding five or more protein transport and trafficking polypeptides.

[00477] In some embodiments, the nucleotide sequences encoding the one or more protein transport and trafficking polypeptides are codon-optimized.

[00478] In some embodiments, one or more protein transport and trafficking polypeptides are overexpressed in the modified host cell. Overexpression may be achieved by increasing the copy number of the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein transport and trafficking polypeptides, e.g., through use of a high copy number expression vector (e.g., a plasmid that exists at 10- 40 copies or about 100 copies per cell) and/or by operably linking the nucleotide sequences encoding the one or more protein transport and trafficking polypeptides to a strong promoter. In some embodiments, the modified host cell has one copy of a heterologous nucleic acid comprising a nucleotide sequence encoding a protein transport and trafficking polypeptide.

In some embodiments, the modified host cell has two copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a protein transport and trafficking polypeptide.

In some embodiments, the modified host cell has three copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a protein transport and trafficking polypeptide.

In some embodiments, the modified host cell has four copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a protein transport and trafficking polypeptide.

In some embodiments, the modified host cell has five copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a protein transport and trafficking polypeptide.

In some embodiments, the modified host cell has five or more copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a protein transport and trafficking polypeptide. Increased copy number of the heterologous nucleic acid and/or codon optimization of the nucleotide sequence may result in an increase in the desired polypeptide activity in the modified host cell.

[00479] Exemplary heterologous nucleic acids disclosed herein may include nucleic acids comprising a nucleotide sequence that encodes a protein transport and trafficking polypeptide, such as, a full-length protein transport and trafficking polypeptide, a fragment of a protein transport and trafficking polypeptide, a variant of a protein transport and trafficking polypeptide, a truncated protein transport and trafficking polypeptide, or a fusion polypeptide that has at least one activity of a protein transport and trafficking polypeptide.

In some embodiments, the nucleotide sequence is codon-optimized.

[00480] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein transport and trafficking polypeptides, the nucleotide sequences encoding the one or more protein transport and trafficking polypeptides are selected from the group consisting of nucleotide sequences set forth in SEQ ID NO: 58 (encodes a SEC23 polypeptide), SEQ ID NO:60 (encodes a SEC24 polypeptide), SEQ ID NO:62 (encodes a SEC 13 polypeptide), SEQ ID NO:20 (encodes a BFR2 polypeptide), SEQ ID NO: 154 (encodes a SEC31 polypeptide), SEQ ID NO: 156 (encodes a SARI polypeptide), SEQ ID NO: 158 (encodes a SEC12 polypeptide), SEQ ID NO: 160 (encodes a SEC4 polypeptide), SEQ ID NO: 162 (encodes a SEC 16 polypeptide), SEQ ID NO: 164 (encodes an ERV29 polypeptide), SEQ ID NO: 166 (encodes a SEC7 polypeptide), SEQ ID NO: 168 (encodes a SEC 5 polypeptide), SEQ ID NO: 170 (encodes a SEC22 polypeptide), SEQ ID NO: 172 (encodes a BOS1 polypeptide), SEQ ID NO: l74 (encodes a BET1 polypeptide), SEQ ID NO: 176 (encodes a BUGl polypeptide), SEQ ID NO: 178 (encodes a GRH1 polypeptide), SEQ ID NO: 180 (encodes a USOl polypeptide), SEQ ID NO: l82 (encodes a SEC 17 polypeptide), SEQ ID NO: 184 (encodes a SEC 18 polypeptide), SEQ ID NO 486 (encodes a SSOl polypeptide), SEQ ID NO: 188 (encodes a SEC9 polypeptide), SEQ ID NO: 190 (encodes a SNC2 polypeptide), and SEQ ID NO: 192 (encodes a LDB17 polypeptide).

[00481] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein transport and trafficking polypeptides, the nucleotide sequences encoding the one or more protein transport and trafficking polypeptides are selected from the group consisting of nucleotide sequences set forth in SEQ ID NO: 58 (encodes a SEC23 polypeptide), SEQ ID NO:60 (encodes a SEC24 polypeptide), SEQ ID NO:62 (encodes a SEC13 polypeptide), SEQ ID NO:20 (encodes a BFR2 polypeptide), SEQ ID NO: 154 (encodes a SEC31 polypeptide), SEQ ID NO: 156 (encodes a SAR1 polypeptide), SEQ ID NO: 158 (encodes a SEC12 polypeptide), SEQ ID NO: 160 (encodes a SEC4 polypeptide), SEQ ID NO: 162 (encodes a SEC 16 polypeptide), SEQ ID NO: 164 (encodes an ERV29 polypeptide), SEQ ID NO 66 (encodes a SEC7 polypeptide), SEQ ID NO 468 (encodes a SEC5 polypeptide), SEQ ID NO470 (encodes a SEC22 polypeptide), SEQ ID NO: 172 (encodes a BOS1 polypeptide), SEQ ID N0474 (encodes a BET1 polypeptide), SEQ ID NO: 176 (encodes a BUGl polypeptide), SEQ ID NO: 178 (encodes a GRH1 polypeptide), SEQ ID NO480 (encodes a USOl polypeptide), SEQ ID N0482 (encodes a SEC17 polypeptide), SEQ ID NO 484 (encodes a SEC 18 polypeptide), SEQ ID NO 486 (encodes a SSOl polypeptide), SEQ ID NO: 188 (encodes a SEC9 polypeptide), SEQ ID NO490 (encodes a SNC2 polypeptide), and SEQ ID NO 492 (encodes a LDB17 polypeptide), or a codon degenerate nucleotide sequence of any of the foregoing.

[00482] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein transport and trafficking polypeptides, the nucleotide sequences encoding the one or more protein transport and trafficking polypeptides are selected from the group consisting of nucleotide sequences having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:58 (encodes a SEC23 polypeptide), SEQ ID NO:60 (encodes a SEC24 polypeptide), SEQ ID NO:62 (encodes a SEC13 polypeptide), SEQ ID NO:20 (encodes a BFR2 polypeptide), SEQ ID NO: 154 (encodes a SEC31 polypeptide), SEQ ID NO: 156 (encodes a SAR1 polypeptide), SEQ ID NO: 158 (encodes a SEC 12 polypeptide), SEQ ID NO: 160 (encodes a SEC4 polypeptide), SEQ ID NO: 162 (encodes a SEC16 polypeptide), SEQ ID NO: 164 (encodes an ERV29 polypeptide), SEQ ID NO: 166 (encodes a SEC7 polypeptide), SEQ ID NO: 168 (encodes a SEC5 polypeptide), SEQ ID NO: 170 (encodes a SEC22 polypeptide), SEQ ID NO: 172 (encodes a BOSl polypeptide), SEQ ID NO:l74 (encodes a BETl polypeptide), SEQ ID NO: 176 (encodes a BUG1 polypeptide), SEQ ID NO: 178 (encodes a GRH1 polypeptide), SEQ ID NO: 180 (encodes a USOl polypeptide), SEQ ID NO: 182 (encodes a SEC17 polypeptide), SEQ ID NO: 184 (encodes a SEC18 polypeptide), SEQ ID NO: 186 (encodes a SSOl polypeptide), SEQ ID NO: 188 (encodes a SEC9 polypeptide), SEQ ID NO: l90 (encodes a SNC2 polypeptide), and SEQ ID NO: l92 (encodes a LDBl7 polypeptide).

[00483] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more protein transport and trafficking polypeptides, the one or more protein transport and trafficking polypeptides are encoded by nucleotide sequences selected from the group consisting of nucleotide sequences set forth in SEQ ID NO:58 (encodes a SEC23 polypeptide), SEQ ID NO: 60 (encodes a SEC24 polypeptide), SEQ ID NO: 62 (encodes a SEC 13 polypeptide),

SEQ ID NO:20 (encodes a BFR2 polypeptide), SEQ ID NO: 154 (encodes a SEC31 polypeptide), SEQ ID NO: 156 (encodes a SAR1 polypeptide), SEQ ID NO: 158 (encodes a SEC12 polypeptide), SEQ ID NO: 160 (encodes a SEC4 polypeptide), SEQ ID NO: 162 (encodes a SEC16 polypeptide), SEQ ID NO: 164 (encodes an ERV29 polypeptide), SEQ ID NO: 166 (encodes a SEC7 polypeptide), SEQ ID NO: 168 (encodes a SEC5 polypeptide),

SEQ ID NO: 170 (encodes a SEC22 polypeptide), SEQ ID NO: 172 (encodes a BOS1 polypeptide), SEQ ID NO: 174 (encodes a BET1 polypeptide), SEQ ID NO: 176 (encodes a BUG1 polypeptide), SEQ ID NO: 178 (encodes a GRH1 polypeptide), SEQ ID NO: 180 (encodes a USOl polypeptide), SEQ ID NO: 182 (encodes a SEC 17 polypeptide), SEQ ID NO: l84 (encodes a SEC18 polypeptide), SEQ ID NO: l86 (encodes a SSOl polypeptide), SEQ ID NO: l88 (encodes a SEC9 polypeptide), SEQ ID NO: l90 (encodes a SNC2 polypeptide), and SEQ ID NO: 192 (encodes a LDB17 polypeptide). Signal Peptidase or Signal Peptidase Complex Polypeptides

[00484] Modified host cells of the disclosure may comprise one or more modifications to modulate the expression of one or more signal peptidase or signal peptidase complex polypeptides. The one or more modifications to modulate the expression of one or more signal peptidase or signal peptidase complex polypeptides may include introducing into a host cell one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more signal peptidase or signal peptidase complex polypeptides and/or deletion or downregulation of one or more genes encoding one or more signal peptidase or signal peptidase complex polypeptides in a host cell. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more signal peptidase or signal peptidase complex polypeptides, resulting in expression or overexpression of the one or more signal peptidase or signal peptidase complex polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more signal peptidase or signal peptidase complex polypeptides, reducing or eliminating the expression of the one or more signal peptidase or signal peptidase complex polypeptides.

[00485] Exemplary signal peptidase or signal peptidase complex polypeptides disclosed herein may also include a full-length signal peptidase or signal peptidase complex polypeptide, a fragment of a signal peptidase or signal peptidase complex polypeptide, a variant of a signal peptidase or signal peptidase complex polypeptide, a truncated signal peptidase or signal peptidase complex polypeptide, or a fusion polypeptide that has at least one activity of a signal peptidase or signal peptidase complex polypeptide.

[00486] Modified host cells of the disclosure may comprise one or more modifications to modulate the expression of one or more signal peptidase or signal peptidase complex polypeptides including, but not limited to, signal peptidase complex polypeptides (e.g., a SPC1 polypeptide, a SPC2 polypeptide, a SCP3 polypeptide, or a SPC11 polypeptide) and signal peptidase polypeptides (e.g., a KEX2 polypeptide).

[00487] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more signal peptidase or signal peptidase complex polypeptides, the one or more signal peptidase or signal peptidase complex polypeptides are selected from the group consisting of: a KEX2 polypeptide, a SPC1 polypeptide, a SPC2 polypeptide, a SPC3 polypeptide, and a SEC11 polypeptide. [00488] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more signal peptidase or signal peptidase complex polypeptides, the one or more signal peptidase or signal peptidase complex polypeptides are selected from the group consisting of signal peptidase or signal peptidase complex polypeptides comprising the amino acid sequences set forth in SEQ ID NO:57 (a KEX2 polypeptide), SEQ ID NO: 147 (a SPC1 polypeptide), SEQ ID N0449 (a SPC2 polypeptide), SEQ ID NO: l5l (a SPC3 polypeptide), and SEQ ID NO: 153 (a SEC11 polypeptide).

[00489] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more signal peptidase or signal peptidase complex polypeptides, the one or more signal peptidase or signal peptidase complex polypeptides are selected from the group consisting of signal peptidase or signal peptidase complex polypeptides comprising the amino acid sequences set forth in SEQ ID NO:57 (a KEX2 polypeptide), SEQ ID NO: 147 (a SPC1 polypeptide), SEQ ID N0449 (a SPC2 polypeptide), SEQ ID NO: 151 (a SPC3 polypeptide), and SEQ ID NO: 153 (a SEC11 polypeptide), or a conservatively substituted amino acid sequence of any of the foregoing.

[00490] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more signal peptidase or signal peptidase complex polypeptides, the one or more signal peptidase or signal peptidase complex polypeptides are selected from the group consisting of signal peptidase or signal peptidase complex polypeptides comprising amino acid sequences having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:57 (a KEX2 polypeptide), SEQ ID NO: 147 (a SPC1 polypeptide), SEQ ID NO: 149 (a SPC2 polypeptide), SEQ ID NO: 151 (a SPC3 polypeptide), and SEQ ID NO: 153 (a SEC11 polypeptide).

[00491] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more signal peptidase or signal peptidase complex polypeptides, the one or more signal peptidase or signal peptidase complex polypeptides are selected from the group consisting of a KEX2 polypeptide, a SPC1 polypeptide, a SPC2 polypeptide, a SPC3 polypeptide, and a SEC11 polypeptide.

[00492] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more signal peptidase or signal peptidase complex polypeptides, the one or more signal peptidase or signal peptidase complex polypeptides are selected from the group consisting of signal peptidase or signal peptidase complex polypeptides comprising the amino acid sequences set forth in SEQ ID NO:57 (a KEX2 polypeptide), SEQ ID NO: 147 (a SPC1 polypeptide), SEQ ID NO: 149 (a SPC2 polypeptide), SEQ ID NO: 151 (a SPC3 polypeptide), and SEQ ID NO: 153 (a SEC11 polypeptide).

[00493] In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a signal peptidase or signal peptidase complex polypeptide. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more signal peptidase or signal peptidase complex polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding two or more signal peptidase or signal peptidase complex polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding three or more signal peptidase or signal peptidase complex polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding four or more signal peptidase or signal peptidase complex polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding five or more signal peptidase or signal peptidase complex polypeptides.

[00494] In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more signal peptidase or signal peptidase complex polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of two or more genes encoding two or more signal peptidase or signal peptidase complex polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of three or more genes encoding three or more signal peptidase or signal peptidase complex polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of four or more genes encoding four or more signal peptidase or signal peptidase complex polypeptides. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of five or more genes encoding five or more signal peptidase or signal peptidase complex polypeptides.

[00495] In some embodiments, the nucleotide sequences encoding the one or more signal peptidase or signal peptidase complex polypeptides are codon-optimized.

[00496] In some embodiments, one or more signal peptidase or signal peptidase complex polypeptides are overexpressed in the modified host cell. Overexpression may be achieved by increasing the copy number of the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more signal peptidase or signal peptidase complex polypeptides, e g., through use of a high copy number expression vector (e.g., a plasmid that exists at 10-40 copies or about 100 copies per cell) and/or by operably linking the nucleotide sequences encoding the one or more signal peptidase or signal peptidase complex polypeptides to a strong promoter. In some embodiments, the modified host cell has one copy of a heterologous nucleic acid comprising a nucleotide sequence encoding a signal peptidase or signal peptidase complex polypeptide. In some embodiments, the modified host cell has two copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a signal peptidase or signal peptidase complex polypeptide. In some embodiments, the modified host cell has three copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a signal peptidase or signal peptidase complex polypeptide In some embodiments, the modified host cell has four copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a signal peptidase or signal peptidase complex polypeptide. In some embodiments, the modified host cell has five copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a signal peptidase or signal peptidase complex polypeptide. In some embodiments, the modified host cell has five or more copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a signal peptidase or signal peptidase complex polypeptide. Increased copy number of the heterologous nucleic acid and/or codon optimization of the nucleotide sequence may result in an increase in the desired polypeptide activity in the modified host cell.

[00497] Exemplary heterologous nucleic acids disclosed herein may include nucleic acids comprising a nucleotide sequence that encodes a signal peptidase or signal peptidase complex polypeptide, such as, a full-length signal peptidase or signal peptidase complex polypeptide, a fragment of a signal peptidase or signal peptidase complex polypeptide, a variant of a signal peptidase or signal peptidase complex polypeptide, a truncated signal peptidase or signal peptidase complex polypeptide, or a fusion polypeptide that has at least one activity of a signal peptidase or signal peptidase complex polypeptide. In some embodiments, the nucleotide sequence is codon-optimized.

[00498] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more signal peptidase or signal peptidase complex polypeptides, the nucleotide sequences encoding the one or more signal peptidase or signal peptidase complex polypeptides are selected from the group consisting of nucleotide sequences set forth in SEQ ID NO:56 (encodes a KEX2 polypeptide), SEQ ID NO: 146 (encodes a SPC1 polypeptide), SEQ ID NO: 148 (encodes a SPC2 polypeptide), SEQ ID NO: 150 (encodes a SPC3 polypeptide), and SEQ ID NO: 152 (encodes a SEC11 polypeptide).

[00499] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more signal peptidase or signal peptidase complex polypeptides, the nucleotide sequences encoding the one or more signal peptidase or signal peptidase complex polypeptides are selected from the group consisting of nucleotide sequences set forth in SEQ ID NO:56 (encodes a KEX2 polypeptide), SEQ ID NO: 146 (encodes a SPC1 polypeptide), SEQ ID NO: 148 (encodes a SPC2 polypeptide), SEQ ID NO: 150 (encodes a SPC3 polypeptide), and SEQ ID NO: 152 (encodes a SEC 11 polypeptide), or a codon degenerate nucleotide sequence of any of the foregoing.

[00500] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more signal peptidase or signal peptidase complex polypeptides, the nucleotide sequences encoding the one or more signal peptidase or signal peptidase complex polypeptides are selected from the group consisting of nucleotide sequences having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:56 (encodes a KEX2 polypeptide), SEQ ID NO: 146 (encodes a SPC1 polypeptide), SEQ ID NO: 148 (encodes a SPC2 polypeptide), SEQ ID NO: 150 (encodes a SPC3 polypeptide), and SEQ ID NO: 152 (encodes a SEC11 polypeptide).

[00501] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more signal peptidase or signal peptidase complex polypeptides, the one or more signal peptidase or signal peptidase complex polypeptides are encoded by nucleotide sequences selected from the group consisting of nucleotide sequences set forth in SEQ ID NO:56 (encodes a KEX2 polypeptide), SEQ ID NO: 146 (encodes a SPC1 polypeptide), SEQ ID NO: 148 (encodes a SPC2 polypeptide), SEQ ID NO: 150 (encodes a SPC3 polypeptide), and SEQ ID NO: 152 (encodes a SEC11 polypeptide).

Polypeptides Involved in Unfolded Protein Response (UPR)

[00502] Modified host cells of the disclosure may comprise one or more modifications to modulate the expression of one or more polypeptides involved in unfolded protein response. The one or more modifications to modulate the expression of one or more polypeptides involved in unfolded protein response may include introducing into a host cell one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response and/or deletion or downregulation of one or more genes encoding one or more polypeptides involved in unfolded protein response in a host cell. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response, resulting in expression or overexpression of the one or more polypeptides involved in unfolded protein response. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more polypeptides involved in unfolded protein response, reducing or eliminating the expression of the one or more polypeptides involved in unfolded protein response.

[00503] Exemplary polypeptides involved in unfolded protein response disclosed herein may also include a full-length polypeptide involved in unfolded protein response, a fragment of a polypeptide involved in unfolded protein response, a variant of a polypeptide involved in unfolded protein response, a truncated polypeptide involved in unfolded protein response, or a fusion polypeptide that has at least one activity of a polypeptide involved in unfolded protein response.

[00504] Modified host cells of the disclosure may comprise one or more modifications to modulate the expression of one or more polypeptides involved in unfolded protein response including, but not limited to, lumenal sensor polypeptides (e.g., an IRE1 polypeptide), transcription factor polypeptides (e.g., a HACls polypeptide), and

polypeptides with a similar expression pattern to the HACls polypeptide (e.g., a DED1 polypeptide or a PPQ1 polypeptide). In some embodiments, manipulation of the unfolded protein response might also be achieved in a synthetic fashion, e.g., by fusion of a transcriptional regulatory domain polypeptide, e.g., the VP16 activator or the Gal4 activation domain to a DNA binding domain polypeptide that interacts with genomic unfolded protein response element (UPRE) nucleotide sequences to regulate gene expression. Such a DNA binding domain polypeptide could either be naturally occurring (e.g., found in the HACls polypeptide) or synthetic (e.g., a Zinc finger, TALEN or Cas9 system). Modulating the expression of one or more polypeptides involved in EIPR may prevent degradation of expressed cannabinoid synthase polypeptides.

[00505] In some embodiments, the one or more modifications to modulate the expression of one or more polypeptides involved in unfolded protein response may improve modified host cell viability. In certain such embodiments, the one or more polypeptides involved in unfolded protein response comprise a PPQ1 polypeptide. In some embodiments, the one or more polypeptides involved in unfolded protein response comprise a DED1 polypeptide. Improving modified host cell viability may improve the industrial fermentation process.

[00506] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response, the one or more polypeptides involved in unfolded protein response are selected from the group consisting of: an IRE1 polypeptide, a HACls polypeptide, a DEDl polypeptide, and a PPQl polypeptide. In some embodiments, the IRE1 polypeptide is a fragment IRE1 polypeptide (e.g., missing the first 7 amino acids).

[00507] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response, the one or more polypeptides involved in unfolded protein response are selected from the group consisting of polypeptides involved in unfolded protein response comprising the amino acid sequences set forth in SEQ ID NO:67 (an IRE1 polypeptide), SEQ ID NO:578 (a fragment IRE1 polypeptide), SEQ ID NO:65 (a HACls polypeptide), SEQ ID NO:265 (an PPQ1 polypeptide), and SEQ ID NO:20l (a DEDl polypeptide).

[00508] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response, the one or more polypeptides involved in unfolded protein response are selected from the group consisting of polypeptides involved in unfolded protein response comprising the amino acid sequences set forth in SEQ ID NO:67 (an IRE1 polypeptide), SEQ ID NO:578 (a fragment IRE1 polypeptide), SEQ ID NO:65 (a HACls polypeptide), SEQ ID NO:265 (an PPQ1 polypeptide), and SEQ ID NO:20l (a DED1 polypeptide), or a conservatively substituted amino acid sequence of any of the foregoing.

[00509] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response, the one or more polypeptides involved in unfolded protein response are selected from the group consisting of polypeptides involved in unfolded protein response comprising amino acid sequences having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:67 (an IRE1 polypeptide), SEQ ID NO:578 (a fragment IRE1 polypeptide), SEQ ID NO:65 (a HACls polypeptide), SEQ ID NO:265 (an PPQ1 polypeptide), and SEQ ID NO:20l (a DEDl polypeptide).

[00510] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more polypeptides involved in unfolded protein response, the one or more polypeptides involved in unfolded protein response are selected from the group consisting of an IRE1 polypeptide, a HACls polypeptide, a DED1 polypeptide, and a PPQ1 polypeptide.

[00511] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more polypeptides involved in unfolded protein response, the one or more polypeptides involved in unfolded protein response are selected from the group consisting of polypeptides involved in unfolded protein response comprising the amino acid sequences set forth in SEQ ID NO:67 (an IRE1 polypeptide), SEQ ID NO:578 (a fragment IRE1 polypeptide), SEQ ID NO:65 (a HACls polypeptide), SEQ ID NO:265 (an PPQ1 polypeptide), and SEQ ID NO:20l (a DEDl polypeptide).

[00512] In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a polypeptide involved in unfolded protein response. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding two or more polypeptides involved in unfolded protein response. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding three or more polypeptides involved in unfolded protein response. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding four or more polypeptides involved in unfolded protein response. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding five or more polypeptides involved in unfolded protein response.

[00513] In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more polypeptides involved in unfolded protein response. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of two or more genes encoding two or more polypeptides involved in unfolded protein response. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of three or more genes encoding three or more polypeptides involved in unfolded protein response. In some embodiments, the modified host cells of the disclosure comprise a deletion or

downregulation of four or more genes encoding four or more polypeptides involved in unfolded protein response. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of five or more genes encoding five or more polypeptides involved in unfolded protein response.

[00514] In some embodiments, the nucleotide sequences encoding the one or more polypeptides involved in unfolded protein response are codon-optimized. In some embodiments, the nucleotide sequences encoding the one or more polypeptides involved in unfolded protein response are a spliced form with the intron removed of the nucleotide sequences encoding the one or more polypeptides involved in unfolded protein response (e.g., the nucleotide sequence encoding the HACls polypeptide). In some embodiments, the intron from the native HAC1 locus is excised.

[00515] In some embodiments, one or more polypeptides involved in unfolded protein response are overexpressed in the modified host cell. Over expression may be achieved by increasing the copy number of the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response, e.g., through use of a high copy number expression vector (e.g., a plasmid that exists at 10-40 copies or about 100 copies per cell) and/or by operably linking the nucleotide sequences encoding the one or more polypeptides involved in unfolded protein response to a strong promoter. In some embodiments, the modified host cell has one copy of a

heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in unfolded protein response. In some embodiments, the modified host cell has two copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in unfolded protein response. In some embodiments, the modified host cell has three copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in unfolded protein response. In some embodiments, the modified host cell has four copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in unfolded protein response. In some embodiments, the modified host cell has five copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in unfolded protein response. In some embodiments, the modified host cell has five or more copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in unfolded protein response. Increased copy number of the heterologous nucleic acid and/or codon optimization of the nucleotide sequence may result in an increase in the desired polypeptide activity in the modified host cell.

[00516] Exemplary heterologous nucleic acids disclosed herein may include nucleic acids comprising a nucleotide sequence that encodes a polypeptide involved in unfolded protein response, such as, a full-length polypeptide involved in unfolded protein response, a fragment of a polypeptide involved in unfolded protein response, a variant of a polypeptide involved in unfolded protein response, a truncated polypeptide involved in unfolded protein response, or a fusion polypeptide that has at least one activity of a polypeptide involved in unfolded protein response. In some embodiments, the nucleotide sequence is codon- optimized. In some embodiments, wherein the polypeptide involved in unfolded protein response is an IRE1 polypeptide, the IRE1 polypeptide is a fragment IRE1 polypeptide (e.g., missing the first 7 amino acids).

[00517] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response, the nucleotide sequences encoding the one or more polypeptides involved in unfolded protein response are selected from the group consisting of nucleotide sequences set forth in SEQ ID NO:66 (encodes an IRE1 polypeptide), SEQ ID NO:577 (encodes a fragment IRE1 polypeptide), SEQ ID NO:64 (encodes a HACls polypeptide), SEQ ID NO:264 (encodes a PPQ1 polypeptide), and SEQ ID NO:200 (encodes a DEDl polypeptide).

[00518] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response, the nucleotide sequences encoding the one or more polypeptides involved in unfolded protein response are selected from the group consisting of nucleotide sequences set forth in SEQ ID NO:66 (encodes an IRE1 polypeptide), SEQ ID NO:577 (encodes a fragment IRE1 polypeptide), SEQ ID NO:64 (encodes a HACls polypeptide), SEQ ID NO:264 (encodes a PPQ1 polypeptide), and SEQ ID NO:200 (encodes a DED1 polypeptide), or a codon degenerate nucleotide sequence of any of the foregoing.

[00519] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response, the nucleotide sequences encoding the one or more polypeptides involved in unfolded protein response are selected from the group consisting of nucleotide sequences having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID N0 66 (encodes an IRE1 polypeptide), SEQ ID NO:577 (encodes a fragment IRE1 polypeptide), SEQ ID NO:64 (encodes a HACls polypeptide), SEQ ID NO:264 (encodes a PPQ1 polypeptide), and SEQ ID NO:200 (encodes a DED1 polypeptide).

[00520] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more polypeptides involved in unfolded protein response, the one or more polypeptides involved in unfolded protein response are encoded by nucleotide sequences selected from the group consisting of nucleotide sequences set forth in SEQ ID NO:66 (encodes an IRE1

polypeptide), SEQ ID NO:577 (encodes a fragment IRE1 polypeptide), SEQ ID NO:64 (encodes a HACls polypeptide), SEQ ID NO:264 (encodes a PPQ1 polypeptide), and SEQ ID NO:200 (encodes a DEDl polypeptide). Polypeptides Involved in Endoplasmic Reticulum-Associated Degradation (ERAD)

[00521] Modified host cells of the disclosure may comprise one or more modifications to modulate the expression of one or more polypeptides involved in endoplasmic reticulum- associated degradation (ERAD). The one or more modifications to modulate the expression of one or more polypeptides involved in endoplasmic reticulum-associated degradation (ERAD) may include introducing into a host cell one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in

endoplasmic reticulum-associated degradation (ERAD) and/or deletion or downregulation of one or more genes encoding one or more polypeptides involved in endoplasmic reticulum- associated degradation (ERAD) in a host cell. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in ERAD, resulting in expression or overexpression of the one or more polypeptides involved in ERAD. In some embodiments, the modified host cells of the disclosure comprise a deletion or

downregulation of one or more genes encoding one or more polypeptides involved in ERAD, reducing or eliminating the expression of the one or more polypeptides involved in ERAD.

[00522] Exemplary polypeptides involved in ERAD disclosed herein may also include a full-length polypeptide involved in ERAD, a fragment of a polypeptide involved in ERAD, a variant of a polypeptide involved in ERAD, a truncated polypeptide involved in ERAD, or a fusion polypeptide that has at least one activity of a polypeptide involved in ERAD.

[00523] Modified host cells of the disclosure may comprise one or more modifications to modulate the expression of one or more polypeptides involved in ERAD including, but not limited to, polypeptides capable of recognizing ubiquitinated proteins (e.g., a NPL4 polypeptide) or ubiquitin ligase polypeptides. Modulating the expression of one or more polypeptides involved in ERAD may prevent degradation of expressed cannabinoid synthase polypeptides. In some embodiments, the one or more modifications to modulate the expression of one or more polypeptides involved in ERAD may improve modified host cell viability. In certain such embodiments, the one or more polypeptides involved in ERAD comprise a NPL4 polypeptide. Improving modified host cell viability may improve the industrial fermentation process.

[00524] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in ERAD, the one or more polypeptides involved in ERAD is a NPL4 polypeptide.

[00525] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in ERAD, the one or more polypeptides involved in ERAD is a NPL4 polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 195.

[00526] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in ERAD, the one or more polypeptides involved in ERAD is a NPL4 polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 195, or a conservatively substituted amino acid sequence thereof.

[00527] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in ERAD, the one or more polypeptides involved in ERAD is a NPL4 polypeptide comprising an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO: 195.

[00528] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more polypeptides involved in ERAD, the one or more polypeptides involved in ERAD is a NPL4 polypeptide.

[00529] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more polypeptides involved in ERAD, the one or more polypeptides involved in ERAD is a NPL4 polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 195.

[00530] In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a polypeptide involved in ERAD. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in ERAD. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding two or more polypeptides involved in ERAD. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding three or more polypeptides involved in ERAD. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding four or more polypeptides involved in ERAD. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding five or more polypeptides involved in ERAD.

[00531] In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more polypeptides involved in ERAD. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of two or more genes encoding two or more polypeptides involved in ERAD. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of three or more genes encoding three or more polypeptides involved in ERAD. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of four or more genes encoding four or more polypeptides involved in ERAD. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of five or more genes encoding five or more polypeptides involved in ERAD.

[00532] In some embodiments, the nucleotide sequences encoding the one or more polypeptides involved in ERAD are codon-optimized.

[00533] In some embodiments, one or more polypeptides involved in ERAD are overexpressed in the modified host cell. Overexpression may be achieved by increasing the copy number of the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in ERAD, e.g., through use of a high copy number expression vector (e g., a plasmid that exists at 10-40 copies or about 100 copies per cell) and/or by operably linking the nucleotide sequences encoding the one or more polypeptides involved in ERAD to a strong promoter. In some embodiments, the modified host cell has one copy of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in ERAD. In some embodiments, the modified host cell has two copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in ERAD. In some embodiments, the modified host cell has three copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in ERAD. In some embodiments, the modified host cell has four copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in ERAD. In some embodiments, the modified host cell has five copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in ERAD. In some embodiments, the modified host cell has five or more copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in ERAD. Increased copy number of the heterologous nucleic acid and/or codon optimization of the nucleotide sequence may result in an increase in the desired polypeptide activity in the modified host cell.

[00534] Exemplary heterologous nucleic acids disclosed herein may include nucleic acids comprising a nucleotide sequence that encodes a polypeptide involved in ERAD, such as, a full-length polypeptide involved in ERAD, a fragment of a polypeptide involved in ERAD, a variant of a polypeptide involved in ERAD, a truncated polypeptide involved in ERAD, or a fusion polypeptide that has at least one activity of a polypeptide involved in ERAD. In some embodiments, the nucleotide sequence is codon-optimized.

[00535] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in ERAD, the nucleotide sequences encoding the one or more polypeptides involved in ERAD is a nucleotide sequence encoding a NPL4

polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO: 194.

[00536] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in ERAD, the nucleotide sequences encoding the one or more polypeptides involved in ERAD is a nucleotide sequence encoding a PL4

polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO: 194, or a codon degenerate nucleotide sequence thereof.

[00537] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in ERAD, the nucleotide sequences encoding the one or more polypeptides involved in ERAD is a nucleotide sequence encoding a NPL4

polypeptide, wherein the nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO: 194.

[00538] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more polypeptides involved in ERAD, the one or more polypeptides involved in ERAD are encoded by a nucleotide sequence encoding a NPL4 polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO: 194.

Polypeptides Involved in Protein Translocation into the Endoplasmic Reticulum

[00539] Modified host cells of the disclosure may comprise one or more modifications to modulate the expression of one or more polypeptides involved in protein translocation into the endoplasmic reticulum. The one or more modifications to modulate the expression of one or more polypeptides involved in protein translocation into the endoplasmic reticulum may include introducing into a host cell one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum and/or deletion or downregulation of one or more genes encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum in a host cell. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum, resulting in expression or over expression of the one or more polypeptides involved in protein translocation into the endoplasmic reticulum. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum, reducing or eliminating the expression of the one or more polypeptides involved in protein translocation into the endoplasmic reticulum.

[00540] Exemplary polypeptides involved in protein translocation into the

endoplasmic reticulum disclosed herein may also include a full-length polypeptide involved in protein translocation into the endoplasmic reticulum, a fragment of a polypeptide involved in protein translocation into the endoplasmic reticulum, a variant of a polypeptide involved in protein translocation into the endoplasmic reticulum, a truncated polypeptide involved in protein translocation into the endoplasmic reticulum, or a fusion polypeptide that has at least one activity of a polypeptide involved in protein translocation into the endoplasmic reticulum.

[00541] Modified host cells of the disclosure may comprise one or more modifications to modulate the expression of one or more polypeptides involved in protein translocation into the endoplasmic reticulum including, but not limited to, translocon protein complex polypeptides (e.g., a SEC61 polypeptide, a SBH1 polypeptide, or a SSS1 polypeptide), SRP protein complex polypeptides (e.g., a SRP14 polypeptide, a SRP21 polypeptide, a SRP68 polypeptide, a SRP72 polypeptide, or a SRP54 polypeptide), and SRP receptor polypeptides (e.g., a SRP101 polypeptide or a SRP102 polypeptide). Modified host cells of the disclosure may also comprise one or more modifications to modulate the expression of the SRP protein complex RNA component SCR1.

[00542] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum, the one or more polypeptides involved in protein translocation into the endoplasmic reticulum are selected from the group consisting of: a KAR2 polypeptide, a SEC61 polypeptide, a SBH1 polypeptide, a SSS1 polypeptide, a SRP14 polypeptide, a SRP21 polypeptide, a SRP68 polypeptide, a SRP72 polypeptide, a SEC65 polypeptide, a SRP54 polypeptide, a SEC62 polypeptide, a SEC63 polypeptide, a SEC66 polypeptide, a SEC72 polypeptide, a SRP 101 polypeptide, and a SRP 102 polypeptide. In some embodiments, the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising a nucleotide sequence encoding SRP protein complex RNA component SCR1.

[00543] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum, the one or more polypeptides involved in protein translocation into the endoplasmic reticulum are selected from the group consisting of polypeptides involved in protein translocation into the endoplasmic reticulum comprising the amino acid sequences set forth in SEQ ID NO:23 (a KAR2 polypeptide), SEQ ID NO: 117 (a SEC61 polypeptide), SEQ ID NO: 119 (a SBH1 polypeptide), SEQ ID NO: 121 (a SSSl polypeptide), SEQ ID NO: l23 (a SRP14 polypeptide), SEQ ID NO: 125 (a SRP21 polypeptide), SEQ ID NO: 127 (a SRP68 polypeptide), SEQ ID NO: 129 (a SRP72 polypeptide), SEQ ID NO: 131 (a SEC65 polypeptide), SEQ ID NO: 133 (a SRP54 polypeptide), SEQ ID NO: 135 (a SEC62 polypeptide), SEQ ID NO: 137 (a SEC63 polypeptide), SEQ ID NO: 139 (a SEC66 polypeptide), SEQ ID NO: 141 (a SEC 72 polypeptide), SEQ ID NO: 143 (a SRP101 polypeptide), and SEQ ID NO: 145 (a SRP102 polypeptide).

[00544] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum, the one or more polypeptides involved in protein translocation into the endoplasmic reticulum are selected from the group consisting of polypeptides involved in protein translocation into the endoplasmic reticulum comprising the amino acid sequences set forth in SEQ ID NO:23 (a KAR2 polypeptide), SEQ ID NO: 117 (a SEC61 polypeptide), SEQ ID NO: 119 (a SBH1 polypeptide), SEQ ID NO: 121 (a SSSl polypeptide), SEQ ID NO: l23 (a SRP14 polypeptide), SEQ ID NO: 125 (a SRP21 polypeptide), SEQ ID NO: 127 (a SRP68 polypeptide), SEQ ID NO: 129 (a SRP72 polypeptide), SEQ ID NO: 131 (a SEC65 polypeptide), SEQ ID NO: 133 (a SRP54 polypeptide), SEQ ID NO: 135 (a SEC62 polypeptide), SEQ ID NO: 137 (a SEC63 polypeptide), SEQ ID NO: 139 (a SEC66 polypeptide), SEQ ID NO: 141 (a SEC 72 polypeptide), SEQ ID NO: 143 (a SRP101 polypeptide), and SEQ ID NO: 145 (a SRP102 polypeptide), or a conservatively substituted amino acid sequence of any of the foregoing.

[00545] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum, the one or more polypeptides involved in protein translocation into the endoplasmic reticulum are selected from the group consisting of polypeptides involved in protein translocation into the endoplasmic reticulum comprising amino acid sequences having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:23 (a KAR2 polypeptide), SEQ ID NO: 117 (a SEC61 polypeptide), SEQ ID NO: 119 (a SBH1 polypeptide), SEQ ID NO: 121 (a SSS1 polypeptide), SEQ ID NO: 123 (a SRP14 polypeptide), SEQ ID NO: 125 (a SRP21 polypeptide), SEQ ID NO: 127 (a SRP68 polypeptide), SEQ ID NO: 129 (a SRP72 polypeptide), SEQ ID NO: 131 (a SEC65 polypeptide), SEQ ID NO: 133 (a SRP54 polypeptide), SEQ ID NO: 135 (a SEC62 polypeptide), SEQ ID NO: 137 (a SEC63 polypeptide), SEQ ID NO: 139 (a SEC66 polypeptide), SEQ ID NO: 141 (a SEC 72 polypeptide), SEQ ID NO: 143 (a SRP101 polypeptide), and SEQ ID NO: 145 (a SRP102 polypeptide).

[00546] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum, the one or more polypeptides involved in protein translocation into the endoplasmic reticulum are selected from the group consisting of a KAR2 polypeptide, a SEC61 polypeptide, a SBH1 polypeptide, a SSS1 polypeptide, a SRP14 polypeptide, a SRP21 polypeptide, a SRP68 polypeptide, a SRP72 polypeptide, a SEC65 polypeptide, a SRP54 polypeptide, a SEC62 polypeptide, a SEC63 polypeptide, a SEC66 polypeptide, a SEC72 polypeptide, a SRP101 polypeptide, and a SRP102 polypeptide.

[00547] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum, the one or more polypeptides involved in protein translocation into the endoplasmic reticulum are selected from the group consisting of polypeptides involved in protein translocation into the endoplasmic reticulum comprising the amino acid sequences set forth in SEQ ID NO:23 (a KAR2 polypeptide), SEQ ID NO: 117 (a SEC 61 polypeptide), SEQ ID NO: 119 (a SBH1 polypeptide), SEQ ID NO: 121 (a SSS1 polypeptide), SEQ ID NO: 123 (a SRP14 polypeptide), SEQ ID NO: 125 (a SRP21 polypeptide), SEQ ID NO: 127 (a SRP68 polypeptide), SEQ ID NO: 129 (a SRP72 polypeptide), SEQ ID NO: 131 (a SEC65 polypeptide), SEQ ID NO: 133 (a SRP54 polypeptide), SEQ ID NO: 135 (a SEC62 polypeptide), SEQ ID NO: 137 (a SEC63 polypeptide), SEQ ID NO: 139 (a SEC66 polypeptide), SEQ ID NO: 141 (a SEC 72 polypeptide), SEQ ID NO: 143 (a SRP101 polypeptide), and SEQ ID NO: 145 (a SRP102 polypeptide).

[00548] In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a polypeptide involved in protein translocation into the endoplasmic reticulum. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding two or more polypeptides involved in protein translocation into the endoplasmic reticulum. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding three or more polypeptides involved in protein translocation into the endoplasmic reticulum. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding four or more polypeptides involved in protein translocation into the endoplasmic reticulum. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding five or more polypeptides involved in protein translocation into the endoplasmic reticulum.

[00549] In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of two or more genes encoding two or more polypeptides involved in protein translocation into the endoplasmic reticulum. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of three or more genes encoding three or more polypeptides involved in protein translocation into the endoplasmic reticulum. In some embodiments, the modified host cells of the disclosure comprise a deletion or

downregulation of four or more genes encoding four or more polypeptides involved in protein translocation into the endoplasmic reticulum. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of five or more genes encoding five or more polypeptides involved in protein translocation into the endoplasmic reticulum.

[00550] In some embodiments, the nucleotide sequences encoding the one or more polypeptides involved in protein translocation into the endoplasmic reticulum are codon- optimized.

[00551] In some embodiments, one or more polypeptides involved in protein translocation into the endoplasmic reticulum are overexpressed in the modified host cell. Overexpression may be achieved by increasing the copy number of the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum, e.g., through use of a high copy number expression vector (e.g., a plasmid that exists at 10-40 copies or about 100 copies per cell) and/or by operably linking the nucleotide sequences encoding the one or more polypeptides involved in protein translocation into the endoplasmic reticulum to a strong promoter. In some embodiments, the modified host cell has one copy of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in protein translocation into the endoplasmic reticulum. In some embodiments, the modified host cell has two copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in protein translocation into the endoplasmic reticulum. In some embodiments, the modified host cell has three copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in protein translocation into the endoplasmic reticulum. In some embodiments, the modified host cell has four copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in protein translocation into the endoplasmic reticulum. In some embodiments, the modified host cell has five copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in protein translocation into the endoplasmic reticulum. In some embodiments, the modified host cell has five or more copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in protein translocation into the endoplasmic reticulum. Increased copy number of the heterologous nucleic acid and/or codon optimization of the nucleotide sequence may result in an increase in the desired polypeptide activity in the modified host cell.

[00552] Exemplary heterologous nucleic acids disclosed herein may include nucleic acids comprising a nucleotide sequence that encodes a polypeptide involved in protein translocation into the endoplasmic reticulum, such as, a full-length polypeptide involved in protein translocation into the endoplasmic reticulum, a fragment of a polypeptide involved in protein translocation into the endoplasmic reticulum, a variant of a polypeptide involved in protein translocation into the endoplasmic reticulum, a truncated polypeptide involved in protein translocation into the endoplasmic reticulum, or a fusion polypeptide that has at least one activity of a polypeptide involved in protein translocation into the endoplasmic reticulum. In some embodiments, the nucleotide sequence is codon-optimized.

[00553] In some embodiments, the modified host cells of the disclosure comprise one copy of a heterologous nucleic acid comprising a nucleotide sequence encoding SRP protein complex RNA component SCR1. In some embodiments, the modified host cells of the disclosure comprise one copy of a heterologous nucleic acid comprising a nucleotide sequence encoding SRP protein complex RNA component SCR1. In some embodiments, the modified host cells of the disclosure comprise two copies of a heterologous nucleic acid comprising a nucleotide sequence encoding SRP protein complex RNA component SCR1. In some embodiments, the modified host cells of the disclosure comprise three copies of a heterologous nucleic acid comprising a nucleotide sequence encoding SRP protein complex RNA component SCR1. In some embodiments, the modified host cells of the disclosure comprise four copies of a heterologous nucleic acid comprising a nucleotide sequence encoding SRP protein complex RNA component SCR1. In some embodiments, the modified host cells of the disclosure comprise five or more copies of a heterologous nucleic acid comprising a nucleotide sequence encoding SRP protein complex RNA component SCR1.

[00554] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum, the nucleotide sequences encoding the one or more polypeptides involved in protein translocation into the endoplasmic reticulum are selected from the group consisting of nucleotide sequences set forth in SEQ ID NO:22 (encodes a KAR2 polypeptide), SEQ ID NO: 116 (encodes a SEC61 polypeptide), SEQ ID NO: 118 (encodes a SBH1 polypeptide), SEQ ID NO: 120 (encodes a SSS1 polypeptide), SEQ ID NO: 122 (encodes a SRP14 polypeptide), SEQ ID NO: 124 (encodes a SRP21 polypeptide), SEQ ID NO: 126 (encodes a SRP68 polypeptide), SEQ ID NO: 128 (encodes a SRP72 polypeptide), SEQ ID NO: 130 (encodes a SEC65 polypeptide), SEQ ID NO: 132 (encodes a SRP54 polypeptide), SEQ ID NO: 134 (encodes a SEC62 polypeptide), SEQ ID NO: 136 (encodes a SEC63 polypeptide), SEQ ID NO: 138 (encodes a SEC66 polypeptide), SEQ ID NO: 140 (encodes a SEC72 polypeptide), SEQ ID NO: 142 (encodes a SRP101 polypeptide), and SEQ ID NO: 144 (encodes a SRP102 polypeptide). In some embodiments, the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising a nucleotide sequence encoding SRP protein complex RNA component SCR1, wherein the nucleotide sequence is that set forth in SEQ ID NO:222.

[00555] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum, the nucleotide sequences encoding the one or more polypeptides involved in protein translocation into the endoplasmic reticulum are selected from the group consisting of nucleotide sequences set forth in SEQ ID NO:22 (encodes a KAR2 polypeptide), SEQ ID NO: 116 (encodes a SEC61 polypeptide), SEQ ID NO: 118 (encodes a SBH1 polypeptide), SEQ ID NO: 120 (encodes a SSS1 polypeptide), SEQ ID NO: 122 (encodes a SRP14 polypeptide), SEQ ID NO: 124 (encodes a SRP21 polypeptide), SEQ ID NO: 126 (encodes a SRP68 polypeptide), SEQ ID NO: 128 (encodes a SRP72 polypeptide), SEQ ID NO: 130 (encodes a SEC65 polypeptide), SEQ ID NO: 132 (encodes a SRP54 polypeptide), SEQ ID NO: 134 (encodes a SEC62 polypeptide), SEQ ID NO: 136 (encodes a SEC63 polypeptide), SEQ ID NO: 138 (encodes a SEC66 polypeptide), SEQ ID NO: 140 (encodes a SEC72 polypeptide), SEQ ID NO: 142 (encodes a SRP101 polypeptide), and SEQ ID NO: 144 (encodes a SRP102 polypeptide), or a codon degenerate nucleotide sequence of any of the foregoing. In some embodiments, the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising a nucleotide sequence encoding SRP protein complex RNA component SCR1, wherein the nucleotide sequence is that set forth in SEQ ID NO:222, or a codon degenerate nucleotide sequence thereof.

[00556] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum, the nucleotide sequences encoding the one or more polypeptides involved in protein translocation into the endoplasmic reticulum are selected from the group consisting of nucleotide sequences having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:22 (encodes a KAR2 polypeptide), SEQ ID NO: l 16 (encodes a SEC61 polypeptide), SEQ ID NO: 118 (encodes a SBH1 polypeptide), SEQ ID NO: 120 (encodes a SSS1 polypeptide), SEQ ID NO: 122 (encodes a SRP14 polypeptide), SEQ ID NO: 124 (encodes a SRP21 polypeptide), SEQ ID NO: 126 (encodes a SRP68 polypeptide), SEQ ID NO: 128 (encodes a SRP72 polypeptide), SEQ ID NO: 130 (encodes a SEC65 polypeptide), SEQ ID NO: 132 (encodes a SRP54 polypeptide), SEQ ID NO: 134 (encodes a SEC62 polypeptide), SEQ ID NO: 136 (encodes a SEC63 polypeptide), SEQ ID NO: 138 (encodes a SEC66 polypeptide), SEQ ID NO: 140 (encodes a SEC72 polypeptide), SEQ ID NO: 142 (encodes a SRP101 polypeptide), and SEQ ID NO: 144 (encodes a SRP102 polypeptide). In some embodiments, the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising a nucleotide sequence encoding SRP protein complex RNA component SCR1, wherein the nucleotide sequence have at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:222.

[00557] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum, the one or more polypeptides involved in protein translocation into the endoplasmic reticulum are encoded by nucleotide sequences selected from the group consisting of nucleotide sequences set forth in SEQ ID NO:22 (encodes a KAR2 polypeptide), SEQ ID NO: 116 (encodes a SEC61 polypeptide), SEQ ID NO: 118 (encodes a SBH1 polypeptide), SEQ ID NO:120 (encodes a SSS1 polypeptide), SEQ ID NO: 122 (encodes a SRP14 polypeptide), SEQ ID NO: 124 (encodes a SRP21 polypeptide), SEQ ID NO: 126 (encodes a SRP68 polypeptide), SEQ ID NO: 128 (encodes a SRP72 polypeptide), SEQ ID NO: 130 (encodes a SEC65 polypeptide), SEQ ID NO: 132 (encodes a SRP54 polypeptide), SEQ ID NO: 134 (encodes a SEC62 polypeptide), SEQ ID NO: 136 (encodes a SEC63 polypeptide), SEQ ID NO: 138 (encodes a SEC66 polypeptide), SEQ ID NO: 140 (encodes a SEC72 polypeptide), SEQ ID NO: 142 (encodes a SRP101 polypeptide), and SEQ ID NO: 144 (encodes a SRP102 polypeptide). In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding SRP protein complex RNA component SCR.1, wherein the SRP protein complex RNA component SCR.1 is encoded by a nucleotide sequence having the nucleotide sequence set forth in SEQ ID N0 222.

Polypeptides Involved in Cell Wall Assembly

[00558] Modified host cells of the disclosure may comprise one or more modifications to modulate the expression of one or more polypeptides involved in cell wall assembly. The one or more modifications to modulate the expression of one or more polypeptides involved in cell wall assembly may include introducing into a host cell one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cell wall assembly and/or deletion or downregulation of one or more genes encoding one or more polypeptides involved in cell wall assembly in a host cell. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cell wall assembly, resulting in expression or overexpression of the one or more polypeptides involved in cell wall assembly. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more polypeptides involved in cell wall assembly, reducing or eliminating the expression of the one or more polypeptides involved in cell wall assembly.

[00559] Exemplary polypeptides involved in cell wall assembly disclosed herein may also include a full-length polypeptide involved in cell wall assembly, a fragment of a polypeptide involved in cell wall assembly, a variant of a polypeptide involved in cell wall assembly, a truncated polypeptide involved in cell wall assembly, or a fusion polypeptide that has at least one activity of a polypeptide involved in cell wall assembly.

[00560] Modified host cells of the disclosure may comprise one or more modifications to modulate the expression of one or more polypeptides involved in cell wall assembly including, but not limited to, cell wall polypeptides involved in beta-glucan assembly (e.g., a KRE1 polypeptide).

[00561] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cell wall assembly, the one or more polypeptides involved in cell wall assembly is a KRE1 polypeptide.

[00562] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cell wall assembly, the one or more polypeptides involved in cell wall assembly is a KRE1 polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 199.

[00563] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cell wall assembly, the one or more polypeptides involved in cell wall assembly is a KRE1 polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 199, or a conservatively substituted amino acid sequence thereof.

[00564] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cell wall assembly, the one or more polypeptides involved in cell wall assembly is a KRE1 polypeptide comprising an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO: 199.

[00565] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more polypeptides involved in cell wall assembly, the one or more polypeptides involved in cell wall assembly is a KREl polypeptide.

[00566] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more polypeptides involved in cell wall assembly, the one or more polypeptides involved in cell wall assembly is a KRE1 polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 199.

[00567] In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a polypeptide involved in cell wall assembly. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cell wall assembly. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding two or more polypeptides involved in cell wall assembly. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding three or more polypeptides involved in cell wall assembly.

In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding four or more polypeptides involved in cell wall assembly. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding five or more polypeptides involved in cell wall assembly.

[00568] In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more polypeptides involved in cell wall assembly. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of two or more genes encoding two or more polypeptides involved in cell wall assembly. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of three or more genes encoding three or more polypeptides involved in cell wall assembly. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of four or more genes encoding four or more polypeptides involved in cell wall assembly. In some embodiments, the modified host cells of the disclosure comprise a deletion or

downregulation of five or more genes encoding five or more polypeptides involved in cell wall assembly.

[00569] In some embodiments, the nucleotide sequences encoding the one or more polypeptides involved in cell wall assembly are codon-optimized.

[00570] In some embodiments, one or more polypeptides involved in cell wall assembly are overexpressed in the modified host cell. Overexpression may be achieved by increasing the copy number of the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cell wall assembly, e.g., through use of a high copy number expression vector (e.g., a plasmid that exists at 10- 40 copies or about 100 copies per cell) and/or by operably linking the nucleotide sequences encoding the one or more polypeptides involved in cell wall assembly to a strong promoter. In some embodiments, the modified host cell has one copy of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in cell wall assembly. In some embodiments, the modified host cell has two copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in cell wall assembly. In some embodiments, the modified host cell has three copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in cell wall assembly. In some embodiments, the modified host cell has four copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in cell wall assembly. In some embodiments, the modified host cell has five copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in cell wall assembly. In some embodiments, the modified host cell has five or more copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in cell wall assembly. Increased copy number of the heterologous nucleic acid and/or codon

optimization of the nucleotide sequence may result in an increase in the desired polypeptide activity in the modified host cell.

[00571] Exemplary heterologous nucleic acids disclosed herein may include nucleic acids comprising a nucleotide sequence that encodes a polypeptide involved in cell wall assembly, such as, a full-length polypeptide involved in cell wall assembly, a fragment of a polypeptide involved in cell wall assembly, a variant of a polypeptide involved in cell wall assembly, a truncated polypeptide involved in cell wall assembly, or a fusion polypeptide that has at least one activity of a polypeptide involved in cell wall assembly. In some embodiments, the nucleotide sequence is codon-optimized.

[00572] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cell wall assembly, the nucleotide sequences encoding the one or more polypeptides involved in cell wall assembly is a nucleotide sequence encoding a KRE1 polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO: 198.

[00573] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cell wall assembly, the nucleotide sequences encoding the one or more polypeptides involved in cell wall assembly is a nucleotide sequence encoding a KRE1 polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO: 198, or a codon degenerate nucleotide sequence thereof.

[00574] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cell wall assembly, the nucleotide sequences encoding the one or more polypeptides involved in cell wall assembly is a nucleotide sequence encoding a KRE1 polypeptide, wherein the nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO: 198.

[00575] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more polypeptides involved in cell wall assembly, the one or more polypeptides involved in cell wall assembly are encoded by a nucleotide sequence encoding a KRE1 polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO: 198.

Polypeptides Involved in Vacuolar Protein Sorting

[00576] Modified host cells of the disclosure may comprise one or more modifications to modulate the expression of one or more polypeptides involved in vacuolar protein sorting. The one or more modifications to modulate the expression of one or more polypeptides involved in vacuolar protein sorting may include introducing into a host cell one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in vacuolar protein sorting and/or deletion or downregulation of one or more genes encoding one or more polypeptides involved in vacuolar protein sorting in a host cell. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in vacuolar protein sorting, resulting in expression or overexpression of the one or more polypeptides involved in vacuolar protein sorting. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more polypeptides involved in vacuolar protein sorting, reducing or eliminating the expression of the one or more polypeptides involved in vacuolar protein sorting.

[00577] Exemplary polypeptides involved in unfolded protein response disclosed herein may also include a full-length polypeptide involved in vacuolar protein sorting, a fragment of a polypeptide involved in vacuolar protein sorting, a variant of a polypeptide involved in vacuolar protein sorting, a truncated polypeptide involved in vacuolar protein sorting, or a fusion polypeptide that has at least one activity of a polypeptide involved in vacuolar protein sorting.

[00578] Modified host cells of the disclosure may comprise one or more modifications to modulate the expression of one or more polypeptides involved in vacuolar protein sorting including, but not limited to, a PEP1 polypeptide and vacuolar proteinase polypeptides (e.g., a PEP4 polypeptide, a PRC1 polypeptide or a PRB1 polypeptide). In some embodiments, the one or more modifications to modulate the expression of one or more polypeptides involved in vacuolar protein sorting may improve modified host cell viability. In certain such embodiments, the one or more polypeptides involved in vacuolar protein sorting comprise a PEP1 polypeptide. Improving modified host cell viability may improve the industrial fermentation process.

[00579] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in vacuolar protein sorting, the one or more polypeptides involved in vacuolar protein sorting are selected from the group consisting of: a PEP1 polypeptide, a PEP4 polypeptide, a PRCl polypeptide, and a PRBl polypeptide.

[00580] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in vacuolar protein sorting, the one or more polypeptides involved in vacuolar protein sorting are selected from the group consisting of polypeptides involved in vacuolar protein sorting comprising the amino acid sequences set forth in SEQ ID NO: 197 (a PEP1 polypeptide), SEQ ID NO:2l3 (a PEP4 polypeptide), SEQ ID NO:229 (a PRCl polypeptide), and SEQ ID NO:23 l (a PRBl polypeptide).

[00581] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in vacuolar protein sorting, the one or more polypeptides involved in vacuolar protein sorting are selected from the group consisting of polypeptides involved in vacuolar protein sorting comprising the amino acid sequences set forth in SEQ ID NO: 197 (a PEP1 polypeptide), SEQ ID NO:2l3 (a PEP4 polypeptide), SEQ ID NO:229 (a PRC1 polypeptide), and SEQ ID NO:23 l (a PRB1 polypeptide), or a conservatively substituted amino acid sequence of any of the foregoing.

[00582] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in vacuolar protein sorting, the one or more polypeptides involved in vacuolar protein sorting are selected from the group consisting of polypeptides involved in vacuolar protein sorting comprising amino acid sequences having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO: 197 (a PEP1 polypeptide), SEQ ID NO:2l3 (a PEP4 polypeptide), SEQ ID NO:229 (a PRCl polypeptide), and SEQ ID NO:23 l (a PRBl polypeptide).

[00583] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more polypeptides involved in vacuolar protein sorting, the one or more polypeptides involved in vacuolar protein sorting are selected from the group consisting of a PEP1 polypeptide, a PEP4 polypeptide, a PRCl polypeptide, and a PRB1 polypeptide.

[00584] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more polypeptides involved in vacuolar protein sorting, the one or more polypeptides involved in vacuolar protein sorting are selected from the group consisting of polypeptides involved in vacuolar protein sorting comprising the amino acid sequences set forth in SEQ ID NO: 197 (a PEP1 polypeptide), SEQ ID NO:2l3 (a PEP4 polypeptide), SEQ ID NO:229 (a PRC1 polypeptide), and SEQ ID NO:23 l (a PRBl polypeptide).

[00585] In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a polypeptide involved in vacuolar protein sorting. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in vacuolar protein sorting. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding two or more polypeptides involved in vacuolar protein sorting. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding three or more polypeptides involved in vacuolar protein sorting. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding four or more polypeptides involved in vacuolar protein sorting. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding five or more polypeptides involved in vacuolar protein sorting.

[00586] In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more polypeptides involved in vacuolar protein sorting. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of two or more genes encoding two or more polypeptides involved in vacuolar protein sorting. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of three or more genes encoding three or more polypeptides involved in vacuolar protein sorting. In some embodiments, the modified host cells of the disclosure comprise a deletion or

downregulation of four or more genes encoding four or more polypeptides involved in vacuolar protein sorting. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of five or more genes encoding five or more polypeptides involved in vacuolar protein sorting.

[00587] In some embodiments, the nucleotide sequences encoding the one or more polypeptides involved in vacuolar protein sorting are codon-optimized. [00588] In some embodiments, one or more polypeptides involved in vacuolar protein sorting are overexpressed in the modified host cell. Overexpression may be achieved by increasing the copy number of the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in vacuolar protein sorting, e.g., through use of a high copy number expression vector (e.g., a plasmid that exists at 10-40 copies or about 100 copies per cell) and/or by operably linking the nucleotide sequences encoding the one or more polypeptides involved in vacuolar protein sorting to a strong promoter. In some embodiments, the modified host cell has one copy of a

heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in vacuolar protein sorting. In some embodiments, the modified host cell has two copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in vacuolar protein sorting. In some embodiments, the modified host cell has three copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in vacuolar protein sorting. In some embodiments, the modified host cell has four copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in vacuolar protein sorting. In some

embodiments, the modified host cell has five copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in vacuolar protein sorting. In some embodiments, the modified host cell has five or more copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in vacuolar protein sorting. Increased copy number of the heterologous nucleic acid and/or codon optimization of the nucleotide sequence may result in an increase in the desired polypeptide activity in the modified host cell.

[00589] Exemplary heterologous nucleic acids disclosed herein may include nucleic acids comprising a nucleotide sequence that encodes a polypeptide involved in vacuolar protein sorting, such as, a full-length polypeptide involved in vacuolar protein sorting, a fragment of a polypeptide involved in vacuolar protein sorting, a variant of a polypeptide involved in vacuolar protein sorting, a truncated polypeptide involved in vacuolar protein sorting, or a fusion polypeptide that has at least one activity of a polypeptide involved in vacuolar protein sorting. In some embodiments, the nucleotide sequence is codon- optimized.

[00590] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in vacuolar protein sorting, the nucleotide sequences encoding the one or more polypeptides involved in vacuolar protein sorting are selected from the group consisting of nucleotide sequences set forth in: SEQ ID NO: 196 (encodes a PEP1 polypeptide), SEQ ID NO:212 (encodes a PEP4 polypeptide), SEQ ID NO:228 (encodes a PRC1 polypeptide), and SEQ ID NO:230 (encodes a PRB l polypeptide).

[00591] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in vacuolar protein sorting, the nucleotide sequences encoding the one or more polypeptides involved in vacuolar protein sorting are selected from the group consisting of nucleotide sequences set forth in SEQ ID NO: 196 (encodes a PEP1 polypeptide), SEQ ID NO:212 (encodes a PEP4 polypeptide), SEQ ID NO:228 (encodes a PRC1 polypeptide), and SEQ ID NO:230 (encodes a PRB l polypeptide), or a codon degenerate nucleotide sequence of any of the foregoing.

[00592] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in vacuolar protein sorting, the nucleotide sequences encoding the one or more polypeptides involved in vacuolar protein sorting are selected from the group consisting of nucleotide sequences having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO: 196 (encodes a PEP1 polypeptide), SEQ ID NO:2l2 (encodes a PEP4 polypeptide), SEQ ID NO:228 (encodes a PRC1 polypeptide), and SEQ ID NO:230 (encodes a PRB1 polypeptide)

[00593] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more polypeptides involved in vacuolar protein sorting, the one or more polypeptides involved in vacuolar protein sorting are encoded by nucleotide sequences selected from the group consisting of nucleotide sequences set forth in SEQ ID NO: 196 (encodes a PEP1 polypeptide), SEQ ID NO:212 (encodes a PEP4 polypeptide), SEQ ID NO:228 (encodes a PRC1 polypeptide), and SEQ ID NO:230 (encodes a PRB l polypeptide).

[00594] In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more of the following genes encoding vacuolar proteinase polypeptides: a PEP4 gene, a PRC1 gene, or a PRB1 gene. In some embodiments, the modified host cells of the disclosure comprise a deletion of one or more of the following genes encoding vacuolar proteinase polypeptides: a PEP4 gene, a PRC1 gene, or a PRB1 gene. In some embodiments, the modified host cells of the disclosure comprise a

downregulation of one or more of the following genes encoding vacuolar proteinase polypeptides: a PEP4 gene, a PRC1 gene, or a PRB1 gene. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of a PEP4 gene. In some embodiments, the modified host cells of the disclosure comprise a deletion of a PEP4 gene. In some embodiments, the modified host cells of the disclosure comprise a downregulation of a PEP4 gene. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of a PRC1 gene. In some embodiments, the modified host cells of the disclosure comprise a deletion of a PRC1 gene. In some embodiments, the modified host cells of the disclosure comprise a downregulation of a PRC1 gene. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of a PRB1 gene. In some embodiments, the modified host cells of the disclosure comprise a deletion of a PRB1 gene. In some embodiments, the modified host cells of the disclosure comprise a downregulation of a PRB 1 gene.

Polypeptides Involved in Lipid Droplet Assembly

[00595] Modified host cells of the disclosure may comprise one or more modifications to modulate the expression of one or more polypeptides involved in lipid droplet assembly. The one or more modifications to modulate the expression of one or more polypeptides involved in lipid droplet assembly may include introducing into a host cell one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in lipid droplet assembly and/or deletion or downregulation of one or more genes encoding one or more polypeptides involved in lipid droplet assembly in a host cell. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in lipid droplet assembly, resulting in expression or overexpression of the one or more polypeptides involved in lipid droplet assembly. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more polypeptides involved in lipid droplet assembly, reducing or eliminating the expression of the one or more polypeptides involved in lipid droplet assembly.

[00596] Exemplary polypeptides involved in lipid droplet assembly disclosed herein may also include a full-length polypeptide involved in lipid droplet assembly, a fragment of a polypeptide involved in lipid droplet assembly, a variant of a polypeptide involved in lipid droplet assembly, a truncated polypeptide involved in lipid droplet assembly, or a fusion polypeptide that has at least one activity of a polypeptide involved in lipid droplet assembly.

[00597] Modified host cells of the disclosure may comprise one or more modifications to modulate the expression of one or more polypeptides involved in lipid droplet assembly including, but not limited to, polypeptides capable of controlling particle size and budding (e.g., a FLDl polypeptide).

[00598] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in lipid droplet assembly, the one or more polypeptides involved in lipid droplet assembly is a FLD1 polypeptide.

[00599] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in lipid droplet assembly, the one or more polypeptides involved in lipid droplet assembly is a FLD1 polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 209.

[00600] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in lipid droplet assembly, the one or more polypeptides involved in lipid droplet assembly is a FLD1 polypeptide comprising the amino acid sequence set forth in SEQ ID NO:209, or a conservatively substituted amino acid sequence thereof.

[00601] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in lipid droplet assembly, the one or more polypeptides involved in lipid droplet assembly is a FLD1 polypeptide comprising an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:209.

[00602] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more polypeptides involved in lipid droplet assembly, the one or more polypeptides involved in lipid droplet assembly is a FLD1 polypeptide.

[00603] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more polypeptides involved in lipid droplet assembly, the one or more polypeptides involved in lipid droplet assembly is a FLD1 polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 209.

[00604] In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a polypeptide involved in lipid droplet assembly. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in lipid droplet assembly. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding two or more polypeptides involved in lipid droplet assembly. In some

embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding three or more polypeptides involved in lipid droplet assembly. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding four or more polypeptides involved in lipid droplet assembly In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding five or more polypeptides involved in lipid droplet assembly.

[00605] In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more polypeptides involved in lipid droplet assembly. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of two or more genes encoding two or more polypeptides involved in lipid droplet assembly. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of three or more genes encoding three or more polypeptides involved in lipid droplet assembly. In some embodiments, the modified host cells of the disclosure comprise a deletion or

downregulation of four or more genes encoding four or more polypeptides involved in lipid droplet assembly. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of five or more genes encoding five or more polypeptides involved in lipid droplet assembly.

[00606] In some embodiments, the nucleotide sequences encoding the one or more polypeptides involved in lipid droplet assembly are codon-optimized.

[00607] In some embodiments, one or more polypeptides involved in lipid droplet assembly are overexpressed in the modified host cell. Overexpression may be achieved by increasing the copy number of the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in lipid droplet assembly, e.g., through use of a high copy number expression vector (e.g., a plasmid that exists at 10- 40 copies or about 100 copies per cell) and/or by operably linking the nucleotide sequences encoding the one or more polypeptides involved in lipid droplet assembly to a strong promoter. In some embodiments, the modified host cell has one copy of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in lipid droplet assembly. In some embodiments, the modified host cell has two copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in lipid droplet assembly. In some embodiments, the modified host cell has three copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in lipid droplet assembly. In some embodiments, the modified host cell has four copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in lipid droplet assembly. In some embodiments, the modified host cell has five copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in lipid droplet assembly. In some embodiments, the modified host cell has five or more copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in lipid droplet assembly. Increased copy number of the heterologous nucleic acid and/or codon optimization of the nucleotide sequence may result in an increase in the desired polypeptide activity in the modified host cell.

[00608] Exemplary heterologous nucleic acids disclosed herein may include nucleic acids comprising a nucleotide sequence that encodes a polypeptide involved in lipid droplet assembly, such as, a full-length polypeptide involved in lipid droplet assembly, a fragment of a polypeptide involved in lipid droplet assembly, a variant of a polypeptide involved in lipid droplet assembly, a truncated polypeptide involved in lipid droplet assembly, or a fusion polypeptide that has at least one activity of a polypeptide involved in lipid droplet assembly. In some embodiments, the nucleotide sequence is codon-optimized. [00609] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in lipid droplet assembly, the nucleotide sequences encoding the one or more polypeptides involved in lipid droplet assembly is a nucleotide sequence encoding a FLD1 polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:208.

[00610] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in lipid droplet assembly, the nucleotide sequences encoding the one or more polypeptides involved in lipid droplet assembly is a nucleotide sequence encoding a FLD1 polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:208, or a codon degenerate nucleotide sequence thereof.

[00611] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in lipid droplet assembly, the nucleotide sequences encoding the one or more polypeptides involved in lipid droplet assembly is a nucleotide sequence encoding a FLDl polypeptide, wherein the nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:208.

[00612] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more polypeptides involved in lipid droplet assembly, the one or more polypeptides involved in lipid droplet assembly are encoded by a nucleotide sequence encoding a FLD1 polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:208.

[00613] In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of a FLD1 gene. In some embodiments, the modified host cells of the disclosure comprise a deletion of a FLD1 gene. In some embodiments, the modified host cells of the disclosure comprise a downregulation of a FLD1 gene. Polypeptides Involved in Regulation of Lipid Metabolism

[00614] Modified host cells of the disclosure may comprise one or more modifications to modulate the expression of one or more polypeptides involved in regulation of lipid metabolism. The one or more modifications to modulate the expression of one or more polypeptides involved in regulation of lipid metabolism may include introducing into a host cell one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in regulation of lipid metabolism and/or deletion or downregulation of one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism in a host cell. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in regulation of lipid metabolism, resulting in expression or overexpression of the one or more polypeptides involved in regulation of lipid metabolism. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism, reducing or eliminating the expression of the one or more polypeptides involved in regulation of lipid metabolism.

[00615] Exemplary polypeptides involved in regulation of lipid metabolism disclosed herein may also include a full-length polypeptide involved in regulation of lipid metabolism, a fragment of a polypeptide involved in regulation of lipid metabolism, a variant of a polypeptide involved in regulation of lipid metabolism, a truncated polypeptide involved in regulation of lipid metabolism, or a fusion polypeptide that has at least one activity of a polypeptide involved in regulation of lipid metabolism.

[00616] Modified host cells of the disclosure may comprise one or more modifications to modulate the expression of one or more polypeptides involved in the regulation of lipid metabolism including, but not limited to, transcriptional regulator polypeptides (e.g., an OPI1 polypeptide).

[00617] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in regulation of lipid metabolism, the one or more polypeptides involved in regulation of lipid metabolism is an OPI1 polypeptide.

[00618] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in regulation of lipid metabolism, the one or more polypeptides involved in regulation of lipid metabolism is an OPI1 polypeptide comprising the amino acid sequence set forth in SEQ ID NO:2l 1.

[00619] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in regulation of lipid metabolism, the one or more polypeptides involved in regulation of lipid metabolism is an OPI1 polypeptide comprising the amino acid sequence set forth in SEQ ID NO:2l 1, or a conservatively substituted amino acid sequence thereof.

[00620] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in regulation of lipid metabolism, the one or more polypeptides involved in regulation of lipid metabolism is an OPI1 polypeptide comprising an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:2l l.

[00621] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism, the one or more polypeptides involved in regulation of lipid metabolism is an OPI1 polypeptide.

[00622] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism, the one or more polypeptides involved in regulation of lipid metabolism is an OPI1 polypeptide comprising the amino acid sequence set forth in SEQ ID NO:2l 1.

[00623] In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a polypeptide involved in regulation of lipid metabolism. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in regulation of lipid metabolism. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding two or more polypeptides involved in regulation of lipid metabolism. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding three or more polypeptides involved in regulation of lipid metabolism. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding four or more polypeptides involved in regulation of lipid metabolism. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding five or more polypeptides involved in regulation of lipid metabolism.

[00624] In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of two or more genes encoding two or more polypeptides involved in regulation of lipid metabolism. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of three or more genes encoding three or more polypeptides involved in regulation of lipid metabolism. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of four or more genes encoding four or more polypeptides involved in regulation of lipid metabolism. In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of five or more genes encoding five or more polypeptides involved in regulation of lipid metabolism.

[00625] In some embodiments, the nucleotide sequences encoding the one or more polypeptides involved in regulation of lipid metabolism are codon-optimized.

[00626] In some embodiments, one or more polypeptides involved in regulation of lipid metabolism are overexpressed in the modified host cell. Overexpression may be achieved by increasing the copy number of the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in regulation of lipid metabolism, e.g., through use of a high copy number expression vector (e g., a plasmid that exists at 10-40 copies or about 100 copies per cell) and/or by operably linking the nucleotide sequences encoding the one or more polypeptides involved in regulation of lipid metabolism to a strong promoter. In some embodiments, the modified host cell has one copy of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in regulation of lipid metabolism. In some embodiments, the modified host cell has two copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in regulation of lipid metabolism. In some embodiments, the modified host cell has three copies of a heterologous nucleic acid comprising a nucleotide sequence encoding polypeptide involved in regulation of lipid metabolism. In some embodiments, the modified host cell has four copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in regulation of lipid metabolism. In some embodiments, the modified host cell has five copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in regulation of lipid metabolism. In some embodiments, the modified host cell has five or more copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in regulation of lipid metabolism. Increased copy number of the heterologous nucleic acid and/or codon optimization of the nucleotide sequence may result in an increase in the desired polypeptide activity in the modified host cell.

[00627] Exemplary heterologous nucleic acids disclosed herein may include nucleic acids comprising a nucleotide sequence that encodes a polypeptide involved in regulation of lipid metabolism, such as, a full-length polypeptide involved in regulation of lipid metabolism, a fragment of a polypeptide involved in regulation of lipid metabolism, a variant of a polypeptide involved in regulation of lipid metabolism, a truncated polypeptide involved in regulation of lipid metabolism, or a fusion polypeptide that has at least one activity of a polypeptide involved in regulation of lipid metabolism. In some embodiments, the nucleotide sequence is codon-optimized.

[00628] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in regulation of lipid metabolism, the nucleotide sequences encoding the one or more polypeptides involved in regulation of lipid metabolism is a nucleotide sequence encoding an OPI1 polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:210.

[00629] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in regulation of lipid metabolism, the nucleotide sequences encoding the one or more polypeptides involved in regulation of lipid metabolism is a nucleotide sequence encoding an OPI1 polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:210, or a codon degenerate nucleotide sequence thereof.

[00630] In some embodiments, wherein the modified host cells of the disclosure comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in regulation of lipid metabolism, the nucleotide sequences encoding the one or more polypeptides involved in regulation of lipid metabolism is a nucleotide sequence encoding an OPI1 polypeptide, wherein the nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:210.

[00631] In some embodiments, wherein the modified host cells of the disclosure comprise a deletion or downregulation of one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism, the one or more polypeptides involved in regulation of lipid metabolism are encoded by a nucleotide sequence encoding an OPI1 polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:2lO.

[00632] In some embodiments, the modified host cells of the disclosure comprise a deletion or downregulation of an OPI1 gene. In some embodiments, the modified host cells of the disclosure comprise a deletion of an OPI1 gene. In some embodiments, the modified host cells of the disclosure comprise a downregulation of an OPI1 gene.

Cannabinoid and Cannabinoid Precursor Biosynthetic Pathway Modifications

[00633] A modified host cell of the present disclosure comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide may also comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cannabinoid or cannabinoid precursor (e.g., geranylpyrophosphate (GPP), prenyl phosphates, olivetolic acid, or hexanoyl-CoA) biosynthesis. In addition to cannabinoid synthase polypeptides, such polypeptides may include, but are not limited to: a geranyl pyrophosphate: olivetolic acid geranyltransferase (GOT) polypeptide, a tetraketide synthase (TKS) polypeptide, an olivetolic acid cyclase (OAC) polypeptide, one or more polypeptides having at least one activity of a polypeptide present in the mevalonate (MEV) pathway (e.g., one or more MEV pathway polypeptides), an acyl-activating enzyme (AAE) polypeptide, a polypeptide that generates GPP (e.g., a geranyl pyrophosphate synthetase (GPPS) polypeptide), a polypeptide that condenses two molecules of acetyl-CoA to generate acetoacetyl-CoA (e.g., an acetoacetyl-CoA thiolase polypeptide), and a pyruvate decarboxylase polypeptide. In some embodiments, the nucleotide sequences encoding the one or more polypeptides involved in cannabinoid or cannabinoid precursor biosynthesis are codon-optimized. [00634] The cannabinoid synthase polypeptides and the polypeptides involved in cannabinoid or cannabinoid precursor biosynthesis and the nucleotide sequences encoding the cannabinoid synthase polypeptides or the polypeptides involved in cannabinoid or cannabinoid precursor biosynthesis may be derived from any suitable source, for example, bacteria, yeast, fungi, algae, human, plant (e.g., Cannabis ), or mouse. In some

embodiments, the disclosure also encompasses orthologous genes encoding the cannabinoid synthase polypeptides or polypeptides involved in cannabinoid or cannabinoid precursor biosynthesis disclosed herein. Exemplary cannabinoid synthase polypeptides or polypeptides involved in cannabinoid or cannabinoid precursor biosynthesis disclosed herein may also include a full-length cannabinoid synthase polypeptide or a full-length polypeptide involved in cannabinoid or cannabinoid precursor biosynthesis, a fragment of a cannabinoid synthase polypeptide or a fragment of a polypeptide involved in cannabinoid or cannabinoid precursor biosynthesis, a variant of a cannabinoid synthase polypeptide or a variant of a polypeptide involved in cannabinoid or cannabinoid precursor biosynthesis, a truncated cannabinoid synthase polypeptide or a truncated polypeptide involved in cannabinoid or cannabinoid precursor biosynthesis, or a fusion polypeptide that has at least one activity of a cannabinoid synthase polypeptide or a fusion polypeptide that has at least one activity of a polypeptide involved in cannabinoid or cannabinoid precursor biosynthesis. The disclosure also provides for nucleotide sequences encoding cannabinoid synthase polypeptides or polypeptides involved in cannabinoid or cannabinoid precursor biosynthesis, such as, a full-length cannabinoid synthase polypeptide or a full-length polypeptide involved in cannabinoid or cannabinoid precursor biosynthesis, a fragment of a cannabinoid synthase polypeptide or a fragment of a polypeptide involved in cannabinoid or cannabinoid precursor biosynthesis, a variant of a cannabinoid synthase polypeptide or a variant of a polypeptide involved in cannabinoid or cannabinoid precursor biosynthesis, a truncated cannabinoid synthase polypeptide or a truncated polypeptide involved in cannabinoid or cannabinoid precursor biosynthesis, or a fusion polypeptide that has at least one activity of a cannabinoid synthase polypeptide or a fusion polypeptide that has at least one activity of a polypeptide involved in cannabinoid or cannabinoid precursor biosynthesis. In some embodiments, the nucleotide sequences encoding the cannabinoid synthase polypeptides or polypeptides involved in cannabinoid or cannabinoid precursor biosynthesis are codon-optimized.

Cannabinoid Synthase Polypeptides

[00635] A modified host cell of the present disclosure may comprise one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide. Different cannabinoid synthase polypeptides can convert CBGA into other cannabinoids, for example, THCA and CBDA. In some embodiments, the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. In some embodiments, the cannabinoid synthase polypeptide is a cannabichromenic acid synthase polypeptide. In some embodiments, the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00636] Cannabinoid synthase polypeptides, secreted polypeptides, have structural features that may hinder expression in modified host cells, such as modified yeast cells. Cannabinoid synthase polypeptides comprise disulfide bonds, numerous glycosylation sites, including A'-glycosylation sites, and a bicovalently attached flavin adenine dinucleotide (FAD) cofactor moiety. Accordingly, reconstituting the activity of cannabinoid synthase polypeptides in a modified host cell, such as a modified yeast cell, can be challenging and unreliable. Often these secreted polypeptides are misfolded or mislocalized, resulting in low expression, polypeptides lacking activity, reduced host cell viability, and/or cell death. As disclosed herein, manipulation of secretory pathway in host cells modified with one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide may improve expression, folding, and enzymatic activity of the cannabinoid synthase polypeptide as well as viability of the modified host cell. In certain such embodiments, the nucleotide sequence encoding a cannabinoid synthase polypeptide is codon-optimized.

[00637] In some embodiments, expression of the cannabinoid synthase polypeptides in a host cell, such as a yeast cell, modified with one or more nucleic acids comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide did not significantly decrease the growth or viability of the modified host cell compared to an unmodified host cell. In some embodiments, a culture of modified host cells of the disclosure comprising one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide has a cell density that is at least 25% or greater, at least 30% or greater, at least 35% or greater, at least 40% or greater, at least 45% or greater, at least 50% or greater, at least 55% or greater, at least 60% or greater, at least 65% or greater, at least 70% or greater, at least 75% or greater, at least 80% or greater, at least 85% or greater at least 90% or greater, at least 95% or greater, at least 100% or greater, at least 110% or greater, at least 120% or greater, at least 130% or greater, at least 140% or greater, or at least 150% or greater than the cell density of a culture of unmodified control host cells grown for the same period, in the same culture medium, and under the same culture conditions.

[00638] Exemplary cannabinoid synthase polypeptides disclosed herein may include a fragment of a cannabinoid synthase polypeptide, a full-length cannabinoid synthase polypeptide, a variant of a cannabinoid synthase polypeptide, a truncated cannabinoid synthase polypeptide, or a fusion polypeptide that has at least one activity of a cannabinoid synthase polypeptide.

[00639] Exemplary heterologous nucleic acids disclosed herein may include nucleic acids comprising a nucleotide sequence that encodes a cannabinoid synthase polypeptide, such as, a fragment of a cannabinoid synthase polypeptide, a variant of a cannabinoid synthase polypeptide, a full-length cannabinoid synthase polypeptide, a truncated cannabinoid synthase polypeptide, or a fusion polypeptide that has at least one activity of a cannabinoid synthase polypeptide. In some embodiments, the nucleotide sequence is codon- optimized.

[00640] In some embodiments, a cannabinoid synthase polypeptide is a

tetrahydrocannabinolic acid synthase (THCAS) polypeptide. THCAS polypeptides can catalyze the conversion of cannabigerolic acid to THCA. Exemplary THCAS polypeptides disclosed herein may include a fragment of a THCAS polypeptide, a full-length THCAS polypeptide, a variant of a THCAS polypeptide, a truncated THCAS polypeptide, or a fusion polypeptide that has at least one activity of a THCAS polypeptide.

[00641] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a THCAS polypeptide, wherein the THCAS polypeptide comprises the amino acid sequence set forth in SEQ ID NO:239. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a THCAS polypeptide, wherein the THCAS polypeptide comprises the amino acid sequence set forth in SEQ ID NO:239, or a conservatively substituted amino acid sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a THCAS polypeptide, wherein the THCAS polypeptide comprises an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, or at least 75% amino acid sequence identity to SEQ ID NO:239. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a THCAS polypeptide, wherein the THCAS polypeptide comprises an amino acid sequence having at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% amino acid sequence identity to SEQ ID NO:239. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a THCAS polypeptide, wherein the THCAS polypeptide comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:239.

[00642] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a THCAS polypeptide, wherein the THCAS polypeptide is a variant THCAS polypeptide with ant least one amino acid substitution compared to a THCAS polypeptide with an amino acid sequence set forth in SEQ ID NO:239. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a variant THCAS polypeptide, wherein the variant THCAS polypeptide comprises an amino acid sequence set forth in SEQ ID NO:607, SEQ ID NO:609, SEQ ID NO:6l 1, SEQ ID NO:613, SEQ ID NO:615, SEQ ID NO:617, SEQ ID NO:6l9, SEQ ID NO:621, SEQ ID NO:623, SEQ ID NO:625, SEQ ID NO:627, SEQ ID NO:629, SEQ ID NO:631, SEQ ID N0 633, SEQ ID N0 635, SEQ ID N0 637, SEQ ID NO:639, SEQ ID NO:641, SEQ ID NO:643, SEQ ID NO:645, SEQ ID NO:647, SEQ ID NO:649, SEQ ID NO:651, SEQ ID NO:653, SEQ ID NO:655, SEQ ID NO:657, SEQ ID NO:659, SEQ ID NO:661, SEQ ID N0 663, SEQ ID N0 665, SEQ ID N0 667, SEQ ID NO:669, SEQ ID NO:671, SEQ ID NO:673, SEQ ID NO:675, SEQ ID NO:677, SEQ ID NO:679, SEQ ID NO:681, SEQ ID NO:683, SEQ ID NO:685, SEQ ID NO:687, SEQ ID NO:689, SEQ ID NO:691, SEQ ID NO:693, SEQ ID NO:695, SEQ ID NO:697, SEQ ID NO:699, SEQ ID NO:701, SEQ ID NO:703, SEQ ID NO:705, SEQ ID NO:707, SEQ ID NO:709, SEQ ID NO:7l 1, SEQ ID NO:713, SEQ ID NO:715, SEQ ID NO:717, SEQ ID NO:7l9, SEQ ID NO:721, SEQ ID NO:723, SEQ ID NO:725, SEQ ID NO:727, SEQ ID NO:729, SEQ ID NO:731, SEQ ID NO:733, SEQ ID NO:735, SEQ ID NO:737, SEQ ID NO:739, SEQ ID NO:741, or SEQ ID NO: 743.

[00643] The THCAS polypeptide may include a modified THCAS polypeptide with an N-terminal truncation to remove the native signal sequence polypeptide. For example, in some embodiments, the THCAS polypeptide lacks N-terminal amino acids 1-28 of the amino acid sequence set forth in SEQ ID NO:239. In certain such embodiments, the THCAS polypeptide may comprise a signal sequence polypeptide different than that of the native signal sequence polypeptide.

[00644] Exemplary heterologous nucleic acids disclosed herein may include nucleic acids comprising a nucleotide sequence that encodes a THCAS polypeptide, such as, a fragment of a THCAS polypeptide, a variant of a THCAS polypeptide, a full-length THCAS polypeptide, a truncated THCAS polypeptide, or a fusion polypeptide that has at least one activity of a THCAS polypeptide. The disclosure also provides nucleic acids comprising a nucleotide sequence encoding a THCAS polypeptide lacking a stop codon. Nucleic acids comprising a nucleotide sequence encoding THCAS polypeptides lacking the stop codon may be useful for expressing said polypeptides in a construct comprising T2A elements. In some embodiments, the nucleic acid may comprise a nucleotide sequence encoding a truncated THCAS polypeptide with an N-terminal truncation to remove the native signal sequence polypeptide. In some embodiments, the nucleic acid comprising a nucleotide sequence encoding a truncated THCAS polypeptide also comprises a nucleotide sequence encoding a signal sequence polypeptide. In certain such embodiments, after transcription and translation, the resulting THCAS polypeptide is modified with the signal sequence polypeptide. In some embodiments, the nucleotide sequence is codon-optimized.

[00645] In some embodiments, the THCAS polypeptide is overexpressed in the modified host cell. Overexpression may be achieved by increasing the copy number of the one or more heterologous nucleic acids comprising a nucleotide sequence encoding the THCAS polypeptide, e.g., through use of a high copy number expression vector (e.g., a plasmid that exists at 10-40 copies or about 100 copies per cell) and/or by operably linking the nucleotide sequence encoding the THCAS polypeptide to a strong promoter. In some embodiments, the modified host cell has one copy of a heterologous nucleic acid comprising a nucleotide sequence encoding the THCAS polypeptide. In some embodiments, the modified host cell has two copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the THCAS polypeptide. In some embodiments, the modified host cell has three copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the THCAS polypeptide. In some embodiments, the modified host cell has four copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the THCAS polypeptide. In some embodiments, the modified host cell has five copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the THCAS polypeptide. In some embodiments, the modified host cell has six copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the THCAS polypeptide. In some embodiments, the modified host cell has seven copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the THCAS polypeptide. In some embodiments, the modified host cell has eight copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the THCAS polypeptide. In some embodiments, the modified host cell has eight or more copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the THCAS polypeptide. Increased copy number of the heterologous nucleic acid and/or codon optimization of the nucleotide sequence may result in an increase in the desired enzyme catalytic activity in the modified host cell.

[00646] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence is that set forth in SEQ ID NO:238, SEQ ID NO:27l, SEQ ID NO:272. SEQ ID NO:273, or SEQ ID NO:274. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence is that set forth in SEQ ID NO:238, SEQ ID NO:27l, SEQ ID NO:272. SEQ ID N0 273, or SEQ ID NO:274, or a codon degenerate nucleotide sequence of any of the foregoing. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:238, SEQ ID NO:271, SEQ ID NO:272. SEQ ID NO:273, or SEQ ID NO:274. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:238, SEQ ID NO:27l, SEQ ID NO:272. SEQ ID NO:273, or SEQ ID NO:274.

[00647] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence is that set forth in SEQ ID NO:238. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence is that set forth in SEQ ID NO:238, or a codon degenerate nucleotide sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:238. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:238.

[00648] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:238. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon- optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon- optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:238. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:238. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:238.

[00649] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence is that set forth in SEQ ID NO:27l . In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence is that set forth in SEQ ID NO:27l, or a codon degenerate nucleotide sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:271. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:271.

[00650] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:27l. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon- optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon- optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:27l. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:271. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:27l.

[00651] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence is that set forth in SEQ ID NO:272. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence is that set forth in SEQ ID NO:272, or a codon degenerate nucleotide sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:272. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:272.

[00652] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:272. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon- optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon- optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:272. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:272. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:272.

[00653] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence is that set forth in SEQ ID NO:273. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence is that set forth in SEQ ID NO:273, or a codon degenerate nucleotide sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:273. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:273.

[00654] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:273. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon- optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon- optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:273. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:273. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:273.

[00655] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence is that set forth in SEQ ID NO:274. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence is that set forth in SEQ ID NO:274, or a codon degenerate nucleotide sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:274. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:274.

[00656] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:274. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon- optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon- optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:274. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:274. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:274.

[00657] In some embodiments, the one or more heterologous nucleic acids may comprise a codon-optimized nucleotide sequence encoding a variant THCAS polypeptide with at least one amino acid substitution compared to a THCAS polypeptide with an amino acid sequence set forth in SEQ ID NO:239. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon- optimized nucleotide sequence encoding a THCAS polypeptide, wherein the THCAS polypeptide is a variant polypeptide, wherein the codon-optimized nucleotide sequence is that set forth in SEQ ID NO:606, SEQ ID NO:608, SEQ ID NO:6lO, SEQ ID NO:6l2, SEQ ID NO:6l4, SEQ ID NO:6l6, SEQ ID NO:6l8, SEQ ID NO:620, SEQ ID NO:622, SEQ ID NO:624, SEQ ID NO:626, SEQ ID NO:628, SEQ ID NO 630, SEQ ID NO:632, SEQ ID

NO:634, SEQ ID NO:636, SEQ ID NO:638, SEQ ID NO 640, SEQ ID NO:642, SEQ ID

NO:644, SEQ ID NO:646, SEQ ID NO:648, SEQ ID NO 650, SEQ ID NO:652, SEQ ID

NO:654, SEQ ID NO:656, SEQ ID NO:658, SEQ ID NO 660, SEQ ID NO:662, SEQ ID

NO:664, SEQ ID NO:666, SEQ ID NO:668, SEQ ID NO 670, SEQ ID NO:672, SEQ ID NO:674, SEQ ID NO:676, SEQ ID NO:678, SEQ ID NO 680, SEQ ID NO:682, SEQ ID

NO:684, SEQ ID NO:686, SEQ ID NO:688, SEQ ID NO 690, SEQ ID NO:692, SEQ ID

NO:694, SEQ ID NO:696, SEQ ID NO:698, SEQ ID N0:700, SEQ ID NO:702, SEQ ID NO:704, SEQ ID NO:706, SEQ ID NO:708, SEQ ID NO:7lO, SEQ ID NO:7l2, SEQ ID NO:7l4, SEQ ID NO:7l6, SEQ ID NO:7l8, SEQ ID NO 720, SEQ ID NO:722, SEQ ID

NO:724, SEQ ID NO:726, SEQ ID NO:728, SEQ ID NO 730, SEQ ID NO:732, SEQ ID

NO:734, SEQ ID NO:736, SEQ ID NO:738, SEQ ID NO 740, or SEQ ID NO:742.

[00658] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a THCAS polypeptide, wherein the THCAS polypeptide is a variant polypeptide, wherein the codon-optimized nucleotide sequence is that set forth in SEQ ID NO:606, SEQ ID NO:608, SEQ ID NO:6lO, SEQ ID NO:6l2, SEQ ID NO:6l4, SEQ ID NO:6l6, SEQ ID NO:6l8, SEQ ID NO:620, SEQ ID NO:622, SEQ ID N0 624, SEQ ID NO:626, SEQ ID

NO:628, SEQ ID NO:630, SEQ ID N0 632, SEQ ID N0 634, SEQ ID NO:636, SEQ ID

NO:638, SEQ ID NO:640, SEQ ID NO:642, SEQ ID N0 644, SEQ ID NO:646, SEQ ID

NO:648, SEQ ID NO:650, SEQ ID NO:652, SEQ ID N0 654, SEQ ID NO:656, SEQ ID

NO:658, SEQ ID NO:660, SEQ ID N0 662, SEQ ID N0 664, SEQ ID NO:666, SEQ ID

NO:668, SEQ ID NO:670, SEQ ID NO:672, SEQ ID N0 674, SEQ ID NO:676, SEQ ID

NO:678, SEQ ID NO:680, SEQ ID NO:682, SEQ ID N0 684, SEQ ID NO:686, SEQ ID

N0 688, SEQ ID NO 690, SEQ ID N0 692, SEQ ID N0 694, SEQ ID N0 696, SEQ ID

NO:698, SEQ ID NO:700, SEQ ID NO:702, SEQ ID NO:704, SEQ ID NO:706, SEQ ID

NO:708, SEQ ID NO:7lO, SEQ ID NO:7l2, SEQ ID N0 714, SEQ ID NO:7l6, SEQ ID

N0 718, SEQ ID NO 720, SEQ ID N0 722, SEQ ID N0 724, SEQ ID N0 726, SEQ ID

NO:728, SEQ ID NO:730, SEQ ID NO:732, SEQ ID N0 734, SEQ ID NO:736, SEQ ID

NO:738, SEQ ID NO:740, or SEQ ID NO:742, or a codon degenerate nucleotide sequence of any of the foregoing.

[00659] In some embodiments, a cannabinoid synthase polypeptide is a

cannabichromenic acid synthase (CBCAS) polypeptide. CBCAS polypeptides can produce cannabichromenic acid. Exemplary CBCAS polypeptides disclosed herein may include a fragment of a CBCAS polypeptide, a full-length CBCAS polypeptide, a variant of a CBCAS polypeptide, a truncated CBCAS polypeptide, or a fusion polypeptide that has at least one activity of a CBCAS polypeptide.

[00660] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a CBCAS polypeptide, wherein the CBCAS polypeptide comprises the amino acid sequence set forth in SEQ ID NO:241. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a CBCAS polypeptide, wherein the CBCAS polypeptide comprises the amino acid sequence set forth in SEQ ID NO:24l, or a conservatively substituted amino acid sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a CBCAS polypeptide, wherein the CBCAS polypeptide comprises an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, or at least 75% amino acid sequence identity to SEQ ID NO:241. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a CBCAS polypeptide, wherein the CBCAS polypeptide comprises an amino acid sequence having at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% amino acid sequence identity to SEQ ID NO:24l . In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a CBCAS polypeptide, wherein the CBCAS polypeptide comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:241.

[00661] The CBCAS polypeptide may include a modified CBCAS polypeptide with an N-terminal truncation to remove the native signal sequence polypeptide. For example, in some embodiments, the CBCAS polypeptide lacks N-terminal amino acids 1-28 of the amino acid sequence set forth in SEQ ID NO:24l. In certain such embodiments, the CBCAS polypeptide may comprise a signal sequence polypeptide different than that of the native signal sequence polypeptide.

[00662] Exemplary heterologous nucleic acids disclosed herein may include nucleic acids comprising a nucleotide sequence that encodes a CBCAS polypeptide, such as, a fragment of a CBCAS polypeptide, a variant of a CBCAS polypeptide, a full-length CBCAS polypeptide, a truncated CBCAS polypeptide, or a fusion polypeptide that has at least one activity of a CBCAS polypeptide. The disclosure also provides nucleic acids comprising a nucleotide sequence encoding a CBCAS polypeptide lacking a stop codon. Nucleic acids comprising a nucleotide sequence encoding CBCAS polypeptides lacking the stop codon may be useful for expressing said polypeptides in a construct comprising T2A elements. In some embodiments, the nucleic acid may comprise a nucleotide sequence encoding a truncated CBCAS polypeptide with an N-terminal truncation to remove the native signal sequence polypeptide. In some embodiments, the nucleic acid comprising a nucleotide sequence encoding a truncated CBCAS polypeptide also comprises a nucleotide sequence encoding a signal sequence polypeptide. In certain such embodiments, after transcription and translation, the resulting CBCAS polypeptide is modified with the signal sequence polypeptide. In some embodiments, the nucleotide sequence is codon-optimized.

[00663] In some embodiments, the CBCAS polypeptide is overexpressed in the modified host cell. Overexpression may be achieved by increasing the copy number of the one or more heterologous nucleic acids comprising a nucleotide sequence encoding the CBCAS polypeptide, e.g., through use of a high copy number expression vector (e.g., a plasmid that exists at 10-40 copies or about 100 copies per cell) and/or by operably linking the nucleotide sequence encoding the CBCAS polypeptide to a strong promoter. In some embodiments, the modified host cell has one copy of a heterologous nucleic acid comprising a nucleotide sequence encoding the CBCAS polypeptide. In some embodiments, the modified host cell has two copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the CBCAS polypeptide. In some embodiments, the modified host cell has three copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the CBCAS polypeptide. In some embodiments, the modified host cell has four copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the CBCAS polypeptide. In some embodiments, the modified host cell has five copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the CBCAS polypeptide. In some embodiments, the modified host cell has six copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the CBCAS polypeptide. In some embodiments, the modified host cell has seven copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the CBCAS polypeptide. In some embodiments, the modified host cell has eight copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the CBCAS polypeptide. In some embodiments, the modified host cell has eight or more copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the CBCAS polypeptide. Increased copy number of the heterologous nucleic acid and/or codon optimization of the nucleotide sequence may result in an increase in the desired enzyme catalytic activity in the modified host cell. [00664] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBCAS polypeptide, wherein the codon-optimized nucleotide sequence is that set forth in SEQ ID NO:240. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBCAS polypeptide, wherein the codon-optimized nucleotide sequence is that set forth in SEQ ID NO:240, or a codon degenerate nucleotide sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBCAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:240. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBCAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:240.

[00665] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBCAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:240. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon- optimized nucleotide sequence encoding a CBCAS polypeptide, wherein the codon- optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:240. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBCAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:240. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBCAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:240.

[00666] In some embodiments, a cannabinoid synthase polypeptide is cannabidiolic acid synthase (CBDAS) polypeptide. CBDAS polypeptides can catalyze the conversion of cannabigerolic acid to cannabidiolic acid (CBDA). Exemplary CBDAS polypeptides disclosed herein may include a full-length CBDAS polypeptide, a fragment of a CBDAS polypeptide, a variant of a CBDAS polypeptide, a truncated CBDAS polypeptide, or a fusion polypeptide that has at least one activity of a CBDAS polypeptide.

[00667] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide comprises the amino acid sequence set forth in SEQ ID NO:9. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide comprises the amino acid sequence set forth in SEQ ID NO:9, or a conservatively substituted amino acid sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide comprises an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, or at least 75% amino acid sequence identity to SEQ ID NO:9. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide comprises an amino acid sequence having at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% amino acid sequence identity to SEQ ID NO:9. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:9.

[00668] The CBDAS polypeptide may include a modified CBDAS polypeptide with an N-terminal truncation to remove the native signal sequence polypeptide. For example, in some embodiments, the CBDAS polypeptide lacks N-terminal amino acids 1-28 of the amino acid sequence set forth in SEQ ID NO:9. In certain such embodiments, the CBDAS polypeptide may comprise a signal sequence polypeptide different than that of the native signal sequence polypeptide. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide is a truncated CBDAS polypeptide comprising the amino acid sequence set forth in SEQ ID NO:243 or SEQ ID NO:245. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide is a truncated CBDAS polypeptide comprising the amino acid sequence set forth in SEQ ID NO:243 or SEQ ID NO:245, or a conservatively substituted amino acid sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide is a truncated CBDAS polypeptide comprising an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, or at least 75% amino acid sequence identity to SEQ ID NO:243 or SEQ ID NO:245. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide is a truncated CBDAS polypeptide comprising an amino acid sequence having at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% amino acid sequence identity to SEQ ID NO:243 or SEQ ID NO:245. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide is a truncated CBDAS polypeptide comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID N0 243 or SEQ ID N0 245

[00669] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide is a truncated CBDAS polypeptide comprising the amino acid sequence set forth in SEQ ID NO:243. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide is a truncated CBDAS polypeptide comprising the amino acid sequence set forth in SEQ ID NO:243, or a conservatively substituted amino acid sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide is a truncated CBDAS polypeptide comprising an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, or at least 75% amino acid sequence identity to SEQ ID NO:243. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide is a truncated CBDAS polypeptide comprising an amino acid sequence having at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% amino acid sequence identity to SEQ ID NO:243. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide is a truncated CBDAS polypeptide comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:243.

[00670] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide is a truncated CBDAS polypeptide comprising the amino acid sequence set forth in SEQ ID NO:245. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide is a truncated CBDAS polypeptide comprising the amino acid sequence set forth in SEQ ID NO:245, or a conservatively substituted amino acid sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide is a truncated CBDAS polypeptide comprising an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, or at least 75% amino acid sequence identity to SEQ ID NO:245. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide is a truncated CBDAS polypeptide comprising an amino acid sequence having at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% amino acid sequence identity to SEQ ID NO:245. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide is a truncated CBDAS polypeptide comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:245.

[00671] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide comprises the amino acid sequence set forth in SEQ ID NO:257. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide comprises the amino acid sequence set forth in SEQ ID NO:257, or a conservatively substituted amino acid sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more

heterologous nucleic acids comprising a nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide comprises an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, or at least 75% amino acid sequence identity to SEQ ID NO:257. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide comprises an amino acid sequence having at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% amino acid sequence identity to SEQ ID NO:257. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:257.

[00672] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide is a variant CBDAS polypeptide with at least one amino acid substitution compared to a CBDAS polypeptide with an amino acid sequence set forth in SEQ ID NO:9. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a variant CBDAS polypeptide, wherein the variant CBDAS polypeptide comprises an amino acid sequence set forth in SEQ ID NO:390, SEQ ID NO:392, SEQ ID NO: 394,

SEQID NO: 396, SEQID NO:398, SEQID N0:400, SEQ ID NO: 402, SEQID NO:404, SEQ ID NO:406, SEQID NO:408, SEQID NO:410, SEQ ID NO:4l2, SEQID NO:4l4, SEQID NO:416, SEQID NO:418, SEQID NO:420, SEQ ID NO: 422, SEQID NO: 424, SEQID NO:426, SEQID NO:428, SEQID NO:430, SEQ ID NO: 432, SEQID NO:434, SEQID NO:436, SEQID NO:438, SEQID NO:440, SEQ ID NO: 442, SEQID NO: 444, SEQID NO:446, SEQID NO:448, SEQID NO:450, SEQ ID NO:452, SEQID NO:454, SEQID NO:456, SEQID NO:458, SEQID NO:460, SEQ ID NO: 462, SEQID NO:464, SEQID NO:466, SEQID NO:468, SEQID NO:470, SEQ ID NO: 472, SEQID NO:474, SEQID NO:476, SEQID NO:478, SEQID NO:480, SEQ ID NO:482, SEQID NO:484, SEQID NO:486, SEQID NO:488, SEQID NO:490, SEQ ID NO: 492, SEQID NO:494, SEQID NO:496, SEQID NO:498, SEQID N0:500, SEQ ID NO: 502, SEQID NO: 504, SEQID NO: 506, SEQID NO:508, SEQID NO:510, SEQ ID NO:5l2, SEQID NO:5l4, SEQID NO:516, SEQID NO:518, SEQID NO:520, SEQ ID NO: 522, SEQID NO: 524, SEQID NO: 526, SEQID NO:528, SEQID NO:530, SEQ ID NO:532, SEQID NO:534, SEQID NO:536, SEQID NO:538, SEQID NO:540, SEQ ID NO: 542, SEQID NO: 544, SEQID NO: 546, SEQID NO:548, SEQID NO:550, SEQ ID NO:552, SEQID NO:554, SEQID NO:556, SEQID NO:558, SEQID NO:560, SEQ ID NO: 562, SEQID NO: 564, SEQID NO: 566, SEQID NO:568, SEQID NO:570, SEQ ID NO 572, SEQID NO: 574, SEQID NO: 580, SEQID NO:582, SEQID NO: 584, SEQ ID NO:586, SEQID NO:588, SEQID NO: 590, SEQID NO:592, SEQID NO: 594, SEQ ID NO: 596, SEQID NO:598, or SEQID N0:600.

[00673] Exemplary heterologous nucleic acids disclosed herein may include nucleic acids comprising a nucleotide sequence that encodes a CBDAS polypeptide, such as, a full- length CBDAS polypeptide, a fragment of a CBDAS polypeptide, a variant of a CBDAS polypeptide, a truncated CBDAS polypeptide, or a fusion polypeptide that has at least one activity of a CBDAS polypeptide. In some embodiments, the nucleotide sequence is codon- optimized.

[00674] In some embodiments, the CBDAS polypeptide is overexpressed in the modified host cell. Overexpression may be achieved by increasing the copy number of the one or more heterologous nucleic acids comprising a nucleotide sequence encoding the

CBDAS polypeptide, e.g., through use of a high copy number expression vector (e.g., a plasmid that exists at 10-40 copies or about 100 copies per cell) and/or by operably linking the nucleotide sequence encoding the CBDAS polypeptide to a strong promoter. In some embodiments, the modified host cell has one copy of a heterologous nucleic acid comprising a nucleotide sequence encoding the CBDAS polypeptide. In some embodiments, the modified host cell has two copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the CBDAS polypeptide. In some embodiments, the modified host cell has three copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the CBDAS polypeptide. In some embodiments, the modified host cell has four copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the CBDAS polypeptide. In some embodiments, the modified host cell has five copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the CBDAS polypeptide. In some embodiments, the modified host cell has six copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the CBDAS polypeptide. In some embodiments, the modified host cell has seven copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the CBDAS polypeptide. In some embodiments, the modified host cell has eight copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the CBDAS polypeptide. In some embodiments, the modified host cell has eight or more copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the CBDAS polypeptide. Increased copy number of the heterologous nucleic acid and/or codon optimization of the nucleotide sequence may result in an increase in the desired enzyme catalytic activity in the modified host cell.

[00675] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the codon-optimized nucleotide sequence is that set forth in SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO: 6, SEQ ID NO: 7, or SEQ ID NO: 8. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon- optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the codon- optimized nucleotide sequence is that set forth in SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8. In some

embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the codon-optimized nucleotide sequence is that set forth in SEQ ID NO: l, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8, or a codon degenerate nucleotide sequence of any of the foregoing. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the codon-optimized nucleotide sequence is that set forth in SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8, or a codon degenerate nucleotide sequence of any of the foregoing. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8 In some

embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8.

[00676] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO: l, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO: l, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO: l, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO: l, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8. [00677] The disclosure provides nucleic acids comprising one or more sequences encoding a CBDAS polypeptide lacking a stop codon. Nucleic acids comprising a nucleotide sequence encoding a CBDAS polypeptide lacking the stop codon may be useful for expressing said polypeptides in a construct comprising T2A elements. In some

embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the codon-optimized nucleotide sequence is that set forth in SEQ ID NO:256, SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO 260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO:263 (each lacking the stop codon). In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the codon-optimized nucleotide sequence is that set forth in SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO:263. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the codon-optimized nucleotide sequence is that set forth in SEQ ID NO:256, SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:261,

SEQ ID NO:262, or SEQ ID NO:263, or a codon degenerate nucleotide sequence of any of the foregoing. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the codon-optimized nucleotide sequence is that set forth in SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO:263, or a codon degenerate nucleotide sequence of any of the foregoing. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:256, SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:261, SEQ ID NO:262, or SEQ ID NO:263. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:261, SEQ ID NO:262, or SEQ ID NO:263. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:256, SEQ ID NO:258,

SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO:263. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO:263.

[00678] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:256, SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO:263. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO:263. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:256, SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO:263. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO:263. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:256, SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO:263. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO:263. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:256, SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO:263. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO: 263.

[00679] The nucleic acids may comprise a nucleotide sequence encoding a truncated CBDAS polypeptide with an N-terminal truncation to remove the native signal sequence polypeptide. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide is a truncated polypeptide, wherein the codon-optimized nucleotide sequence is that set forth in SEQ ID NO:242 or SEQ ID NO:244. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide is a truncated polypeptide, wherein the codon-optimized nucleotide sequence is that set forth in SEQ ID NO:242 or SEQ ID NO:244, or a codon degenerate nucleotide sequence of any of the foregoing. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide is a truncated polypeptide, wherein the codon-optimized nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:242 or SEQ ID NO:244. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide is a truncated

polypeptide, wherein the codon-optimized nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:242 or SEQ ID NO:244. The nucleic acids comprising a nucleotide sequence encoding a truncated CBDAS polypeptide may also comprise a nucleotide sequence encoding a signal sequence polypeptide. In certain such embodiments, after transcription and translation, the resulting CBDAS polypeptide is modified with the signal sequence polypeptide.

[00680] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide is a truncated

polypeptide, wherein the codon-optimized nucleotide sequence has at least 80% sequence identity to SEQ ID NO:242 or SEQ ID NO:244. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon- optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide is a truncated polypeptide, wherein the codon-optimized nucleotide sequence has at least 85% sequence identity to SEQ ID NO:242 or SEQ ID NO:244. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide is a truncated polypeptide, wherein the codon- optimized nucleotide sequence has at least 90% sequence identity to SEQ ID NO:242 or SEQ ID NO:244. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide is a truncated

polypeptide, wherein the codon-optimized nucleotide sequence has at least 95% sequence identity to SEQ ID NO:242 or SEQ ID NO:244.

[00681] In some embodiments, the one or more heterologous nucleic acids may comprise a codon-optimized nucleotide sequence encoding a variant CBDAS polypeptide with at least one amino acid substitution compared to a CBDAS polypeptide with an amino acid sequence set forth in SEQ ID NO:9. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide is a variant polypeptide, wherein the codon-optimized nucleotide sequence is that set forth in SEQ ID NO:389, SEQ ID NO:39l, SEQ ID NO:393, SEQ ID NO:395, SEQ ID NO:397, SEQ ID NO:399, SEQ ID NO:40l, SEQ ID NO:403, SEQ ID NO:405, SEQ ID NO:407, SEQ ID NO:409, SEQIDNO:41l, SEQIDNO:413, SEQIDNO:4l5, SEQIDNO:4l7, SEQ ID NO:419, SEQ ID NO:42l, SEQ ID NO:423, SEQ ID NO:425, SEQ ID NO:427, SEQ ID NO:429, SEQIDNO:43l, SEQIDNO:433, SEQIDNO:435, SEQIDNO:437, SEQ ID NO:439, SEQ ID NO:44l, SEQ ID NO:443, SEQ ID NO:445, SEQ ID NO:447, SEQ ID NO:449, SEQIDNO:45l, SEQIDNO:453, SEQIDNO:455, SEQIDNO:457, SEQ ID NO:459, SEQ ID NO:46l, SEQ ID NO:463, SEQ ID NO:465, SEQ ID NO:467, SEQ ID NO:469, SEQ ID NO:47l, SEQ ID NO:473, SEQ ID NO:475, SEQ ID NO:477, SEQ ID NO:479, SEQIDNO:48l, SEQIDNO:483, SEQIDNO:485, SEQIDNO:487, SEQ ID NO:489, SEQ ID NO:49l, SEQ ID NO:493, SEQ ID NO:495, SEQ ID NO:497, SEQ ID NO:499, SEQ ID NO:50l, SEQ ID NO:503, SEQ ID NO:505, SEQ ID NO:507, SEQ ID NO:509, SEQIDNO:51l, SEQIDNO:513, SEQIDNO:5l5, SEQIDNO:517, SEQ ID NO:519, SEQIDNO:52l, SEQIDNO:523, SEQIDNO:525, SEQIDNO:527, SEQ ID NO: 529, SEQIDNO:53l, SEQIDNO:533, SEQIDNO:535, SEQIDNO:537, SEQ ID NO:539, SEQ ID NO:54l, SEQ ID NO:543, SEQ ID NO:545, SEQ ID NO:547, SEQ ID NO: 549, SEQIDN0551, SEQIDN0553, SEQIDN0555, SEQIDN0557, SEQ ID NO:559, SEQ ID NO:56l, SEQ ID NO:563, SEQ ID NO:565, SEQ ID NO:567, SEQ ID NO:569, SEQIDNO:57l, SEQIDNO:573, SEQIDNO:579, SEQIDNO:581, SEQ ID N0583, SEQ ID N0585, SEQ ID N0587, SEQ ID N0589, SEQ ID N0591, SEQ ID NO:593, SEQ ID NO:595, SEQ ID NO:597, or SEQ ID NO:599.

[00682] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a CBDAS polypeptide, wherein the CBDAS polypeptide is a variant polypeptide, wherein the codon-optimized nucleotide sequence is that set forth in SEQ ID NO:389, SEQ ID NO:391, SEQ ID NO:393, SEQ ID NO:395, SEQ ID NO:397, SEQ ID NO:399, SEQ ID NO:401, SEQ ID NO:403, SEQ ID NO:405, SEQ ID NO:407, SEQ ID NO:409, SEQ ID NO:411, SEQ ID NO:413, SEQIDNO:415, SEQIDNO:417, SEQIDNO:419, SEQ ID NO:421, SEQ ID NO:423, SEQ ID NO:425, SEQ ID N0427, SEQ ID NO:429, SEQ ID NO:431, SEQ ID NO:433, SEQIDNO:435, SEQIDN0437, SEQIDNO:439, SEQ ID NO:441, SEQ ID NO:443, SEQ ID NO:445, SEQ ID NO:447, SEQ ID NO:449, SEQ ID NO:45l, SEQ ID NO:453, SEQ ID NO:455, SEQ ID NO:457, SEQ ID NO:459, SEQ ID NO:46l, SEQ ID NO:463, SEQ ID NO:465, SEQ ID N0 467, SEQ ID NO:469, SEQ ID NO:47l, SEQ ID NO:473, SEQ ID NO:475, SEQ ID N0 477, SEQ ID NO:479, SEQ ID NO:48l, SEQ ID NO:483, SEQ ID NO:485, SEQ ID NO:487, SEQ ID NO:489, SEQ ID NO:49l, SEQ ID NO:493, SEQ ID NO:495, SEQ ID N0 497, SEQ ID NO:499, SEQ ID NO:50l, SEQ ID NO:503, SEQ ID NO:505, SEQ ID NO: 507, SEQ ID NO:509, SEQ ID NO:5l l, SEQ ID NO:5l3, SEQ ID NO:5 l5, SEQ ID NO:5 l7, SEQ ID NO:5l9, SEQ ID NO:52l, SEQ ID NO:523, SEQ ID NO:525, SEQ ID N0 527, SEQ ID NO:529, SEQ ID NO:53 l, SEQ ID NO:533, SEQ ID NO:535, SEQ ID N0 537, SEQ ID NO:539, SEQ ID NO:54l, SEQ ID NO:543, SEQ ID NO:545, SEQ ID N0 547, SEQ ID NO:549, SEQ ID NO:55l, SEQ ID NO:553, SEQ ID N0 555, SEQ ID N0 557, SEQ ID NO:559, SEQ ID NO:56l, SEQ ID NO:563, SEQ ID NO:565, SEQ ID NO: 567, SEQ ID NO:569, SEQ ID NO:57l, SEQ ID NO:573, SEQ ID NO:579, SEQ ID N0 581, SEQ ID NO:583, SEQ ID NO:585, SEQ ID NO:587, SEQ ID NO: 589, SEQ ID N0 591, SEQ ID NO:593, SEQ ID NO:595, SEQ ID NO:597, or SEQ ID NO:599, or a codon degenerate nucleotide sequence of any of the foregoing.

[00683] In some embodiments, at least one of the one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide is operably linked to an inducible promoter. In some embodiments, at least one of the one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide is operably linked to a constitutive promoter.

[00684] In some embodiments of the modified host cells of the disclosure, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide. In certain such embodiments, the one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide comprise a nucleotide sequence encoding a signal sequence polypeptide. In some embodiments, the signal sequence polypeptide is a secretory signal sequence polypeptide. In some embodiments, the secretory signal sequence polypeptide is a native secretory signal sequence polypeptide. In some embodiments, the secretory signal sequence polypeptide is a synthetic secretory signal sequence polypeptide. In some embodiments, the secretory signal sequence polypeptide is an endoplasmic reticulum retention signal sequence polypeptide. In certain such embodiments, the endoplasmic reticulum retention signal sequence polypeptide is a HDEL polypeptide or a

KDEL polypeptide. In some embodiments, the secretory signal sequence polypeptide is a mitochondrial targeting signal sequence polypeptide. In some embodiments, the secretory signal sequence polypeptide is a Golgi targeting signal sequence polypeptide. In some embodiments, the secretory signal sequence polypeptide is a vacuolar localization signal sequence polypeptide. In certain such embodiments, the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide or a PRClt polypeptide. In certain such embodiments, the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide. In some embodiments, the secretory signal sequence polypeptide is a plasma membrane localization signal sequence polypeptide. In some embodiments, the secretory signal sequence polypeptide is a peroxisome targeting signal sequence polypeptide. In some embodiments, the peroxisome targeting signal sequence polypeptide is a PEX8 polypeptide. In some embodiments, the secretory signal sequence polypeptide is a mating factor secretory signal sequence polypeptide (e.g., a MF polypeptide or an evolved MF polypeptide (MFev)). In some embodiments, the signal sequence polypeptide is linked to the N-terminus of the cannabinoid synthase polypeptide.

[00685] In some embodiments of the modified host cells of the disclosure, the cannabinoid synthase polypeptide is a fusion polypeptide with an AGA2t polypeptide. In certain such embodiments, the one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide comprise a nucleotide sequence encoding an AGA2t polypeptide. In certain such embodiments, the modified host cell of the disclosure is modified with one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AGA1 polypeptide. In certain such embodiments, the cannabinoid synthase-AGA2t fusion polypeptide may be displayed on the cell surface of the modified host cell. In some embodiments, the AGA2t polypeptide is linked to the N- terminus of the cannabinoid synthase polypeptide.

[00686] In some embodiments of the modified host cells of the disclosure, the cannabinoid synthase polypeptide is a fusion polypeptide with a GFP polypeptide. In certain such embodiments, the one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide comprise a nucleotide sequence encoding a GFP polypeptide. In some embodiments, the GFP polypeptide is linked to the N- terminus of the cannabinoid synthase polypeptide.

Geranyl Pyrophosphate :Olivetolic Acid Geranyltransferase (GOT) Polypeptides

[00687] A modified host cell of the present disclosure may comprise one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate: olivetolic acid geranyltransferase (GOT) polypeptide. [00688] Exemplary GOT polypeptides disclosed herein may include a full-length GOT polypeptide, a fragment of a GOT polypeptide, a variant of a GOT polypeptide, a truncated GOT polypeptide, or a fusion polypeptide that has at least one activity of a GOT polypeptide. In some embodiments, the GOT polypeptide has aromatic prenyltransferase (PT) activity. In some embodiments, the GOT polypeptide modifies a cannabinoid precursor or a cannabinoid precursor derivative. In certain such embodiments, the GOT polypeptide modifies olivetolic acid or an olivetolic acid derivative.

[00689] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the GOT polypeptide comprises the amino acid sequence set forth in SEQ ID NO:70 or SEQ ID NO:73. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the GOT polypeptide comprises the amino acid sequence set forth in SEQ ID NO:70 or SEQ ID NO:73, or a conservatively substituted amino acid sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the GOT polypeptide comprises an amino acid sequence having at least 65%, at least 70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO 70 or SEQ ID N0 73.

[00690] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the GOT polypeptide is a truncated GOT (tGOT) polypeptide comprising the amino acid sequence set forth in SEQ ID NO:70. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the GOT polypeptide is a truncated GOT (tGOT) polypeptide comprising the amino acid sequence set forth in SEQ ID NO:70, or a conservatively substituted amino acid sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the GOT polypeptide is a truncated GOT (tGOT) polypeptide comprising an amino acid sequence having at least 65%, at least 70%, or at least 75% amino acid sequence identity to SEQ ID NO:70. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the GOT polypeptide is a truncated GOT (tGOT) polypeptide comprising an amino acid sequence having at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% amino acid sequence identity to SEQ ID NO:70. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the GOT polypeptide is a truncated GOT (tGOT) polypeptide comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:70.

[00691] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the GOT polypeptide comprises the amino acid sequence set forth in SEQ ID NO:73. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the GOT polypeptide comprises the amino acid sequence set forth in SEQ ID NO:73, or a conservatively substituted amino acid sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the GOT polypeptide comprises an amino acid sequence having at least 65%, at least 70%, or at least 75% amino acid sequence identity to SEQ ID NO:73. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the GOT polypeptide comprises an amino acid sequence having at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% amino acid sequence identity to SEQ ID NO:73. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the GOT polypeptide comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:73.

[00692] Exemplary heterologous nucleic acids disclosed herein may include nucleic acids comprising a nucleotide sequence that encodes a GOT polypeptide, such as, a full- length GOT polypeptide, a fragment of a GOT polypeptide, a variant of a GOT polypeptide, a truncated GOT polypeptide, or a fusion polypeptide that has at least one activity of a GOT polypeptide. In some embodiments, the nucleotide sequence is codon-optimized.

[00693] In some embodiments, the GOT polypeptide is overexpressed in the modified host cell. Overexpression may be achieved by increasing the copy number of the one or more heterologous nucleic acids comprising a nucleotide sequence encoding the GOT polypeptide, e.g., through use of a high copy number expression vector (e.g., a plasmid that exists at 10-40 copies or about 100 copies per cell) and/or by operably linking the nucleotide sequence encoding the GOT polypeptide to a strong promoter. In some embodiments, the modified host cell has one copy of a heterologous nucleic acid comprising a nucleotide sequence encoding the GOT polypeptide. In some embodiments, the modified host cell has two copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the GOT polypeptide. In some embodiments, the modified host cell has three copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the GOT polypeptide. In some embodiments, the modified host cell has four copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the GOT polypeptide. In some embodiments, the modified host cell has five copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the GOT polypeptide. In some embodiments, the modified host cell has six copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the GOT polypeptide. In some embodiments, the modified host cell has seven copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the GOT polypeptide. In some embodiments, the modified host cell has eight copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the GOT polypeptide. In some embodiments, the modified host cell has eight or more copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the GOT polypeptide. Increased copy number of the heterologous nucleic acid and/or codon optimization of the nucleotide sequence may result in an increase in the desired enzyme catalytic activity in the modified host cell.

[00694] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:68, SEQ ID NO:69, SEQ ID NO:7l, or SEQ ID NO:72. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:68, SEQ ID NO:69, SEQ ID NO:7l, or SEQ ID NO:72, or a codon degenerate nucleotide sequence of any of the foregoing. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:68, SEQ ID NO:69, SEQ ID NO:7l, or SEQ ID NO:72.

[00695] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the GOT polypeptide is a truncated GOT (tGOT) polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:68. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the GOT polypeptide is a truncated GOT (tGOT) polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:68, or a codon degenerate nucleotide sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the GOT polypeptide is a truncated GOT (tGOT) polypeptide, wherein the nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:68. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the GOT polypeptide is a truncated GOT (tGOT) polypeptide, wherein the nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:68.

[00696] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the GOT polypeptide is a truncated GOT (tGOT) polypeptide, wherein nucleotide sequence has at least 80% sequence identity to SEQ ID NO:68. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the GOT polypeptide is a truncated GOT (tGOT) polypeptide, wherein the nucleotide sequence has at least 85% sequence identity to SEQ ID NO:68. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the GOT polypeptide is a truncated GOT (tGOT) polypeptide, wherein the nucleotide sequence has at least 90% sequence identity to SEQ ID NO:68. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the GOT polypeptide is a truncated GOT (tGOT) polypeptide, wherein the nucleotide sequence has at least 95% sequence identity to SEQ ID NO:68.

[00697] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the GOT polypeptide is a truncated GOT (tGOT) polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:69. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the GOT polypeptide is a truncated GOT (tGOT) polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:69, or a codon degenerate nucleotide sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the GOT polypeptide is a truncated GOT (tGOT) polypeptide, wherein the nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:69. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the GOT polypeptide is a truncated GOT (tGOT) polypeptide, wherein the nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:69.

[00698] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the GOT polypeptide is a truncated GOT (tGOT) polypeptide, wherein the nucleotide sequence has at least 80% sequence identity to SEQ ID NO:69. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the GOT polypeptide is a truncated GOT (tGOT) polypeptide, wherein the nucleotide sequence has at least 85% sequence identity to SEQ ID NO:69. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the GOT polypeptide is a truncated GOT (tGOT) polypeptide, wherein the nucleotide sequence has at least 90% sequence identity to SEQ ID NO:69. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the GOT polypeptide is a truncated GOT (tGOT) polypeptide, wherein the nucleotide sequence has at least 95% sequence identity to SEQ ID NO: 69.

[00699] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:7l. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:71, or a codon degenerate nucleotide sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:7l. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:71.

[00700] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the nucleotide sequence has at least 80% sequence identity to SEQ ID NO:7l. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the nucleotide sequence has at least 85% sequence identity to SEQ ID NO:7l. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the nucleotide sequence has at least 90% sequence identity to SEQ ID NO:7l. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the nucleotide sequence has at least 95% sequence identity to SEQ ID NO:7l.

[00701] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:72. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:72, or a codon degenerate nucleotide sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO: 72. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:72.

[00702] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the nucleotide sequence has at least 80% sequence identity to SEQ ID NO:72. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the nucleotide sequence has at least 85% sequence identity to SEQ ID NO:72. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the nucleotide sequence has at least 90% sequence identity to SEQ ID NO:72. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide, wherein the nucleotide sequence has at least 95% sequence identity to SEQ ID NO:72.

NphB Polypeptides

[00703] In some embodiments, a NphB polypeptide is used instead of a GOT polypeptide to generate cannabigerolic acid from GPP and olivetolic acid. A modified host cell of the present disclosure may comprise one or more heterologous nucleic acids comprising a nucleotide sequence encoding a NphB polypeptide.

[00704] Exemplary NphB polypeptides disclosed herein may include a full-length NphB polypeptide, a fragment of a NphB polypeptide, a variant of a NphB polypeptide, a truncated NphB polypeptide, or a fusion polypeptide that has at least one activity of a NphB polypeptide. In some embodiments, the NphB polypeptide has aromatic prenyltransferase (PT) activity. In some embodiments, the NphB polypeptide modifies a cannabinoid precursor or a cannabinoid precursor derivative. In certain such embodiments, the NphB polypeptide modifies olivetolic acid or an olivetolic acid derivative.

[00705] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a NphB polypeptide, wherein the NphB polypeptide comprises the amino acid sequence set forth in SEQ ID NO:576. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a NphB polypeptide, wherein the NphB polypeptide comprises the amino acid sequence set forth in SEQ ID NO:576, or a conservatively substituted amino acid sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a NphB polypeptide, wherein the NphB polypeptide comprises an amino acid sequence having at least 65%, at least 70%, or at least 75% amino acid sequence identity to SEQ ID NO:576. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a NphB polypeptide, wherein the NphB polypeptide comprises an amino acid sequence having at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% amino acid sequence identity to SEQ ID NO:576. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a NphB polypeptide, wherein the NphB polypeptide comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:576.

[00706] Exemplary heterologous nucleic acids disclosed herein may include nucleic acids comprising a nucleotide sequence that encodes a NphB polypeptide, such as, a full- length NphB polypeptide, a fragment of a NphB polypeptide, a variant of a NphB

polypeptide, a truncated NphB polypeptide, or a fusion polypeptide that has at least one activity of a NphB polypeptide. In some embodiments, the nucleotide sequence is codon- optimized.

[00707] In some embodiments, the NphB polypeptide is overexpressed in the modified host cell. Overexpression may be achieved by increasing the copy number of the one or more heterologous nucleic acids comprising a nucleotide sequence encoding the NphB polypeptide, e.g., through use of a high copy number expression vector (e.g., a plasmid that exists at 10-40 copies or about 100 copies per cell) and/or by operably linking the nucleotide sequence encoding the NphB polypeptide to a strong promoter. In some embodiments, the modified host cell has one copy of a heterologous nucleic acid comprising a nucleotide sequence encoding the NphB polypeptide. In some embodiments, the modified host cell has two copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the NphB polypeptide. In some embodiments, the modified host cell has three copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the NphB polypeptide. In some embodiments, the modified host cell has four copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the NphB polypeptide. In some embodiments, the modified host cell has five copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the NphB polypeptide. In some embodiments, the modified host cell has six copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the NphB polypeptide. In some embodiments, the modified host cell has seven copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the NphB polypeptide. In some embodiments, the modified host cell has eight copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the NphB polypeptide. In some embodiments, the modified host cell has eight or more copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the NphB polypeptide. Increased copy number of the heterologous nucleic acid and/or codon optimization of the nucleotide sequence may result in an increase in the desired enzyme catalytic activity in the modified host cell. [00708] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a NphB polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO: 575. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a NphB polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:575, or a codon degenerate nucleotide sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a NphB polypeptide, wherein the nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:575. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a NphB polypeptide, wherein the nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:575.

[00709] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a NphB polypeptide, wherein the nucleotide sequence has at least 80% sequence identity to SEQ ID NO: 575. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a NphB polypeptide, wherein the nucleotide sequence has at least 85% sequence identity to SEQ ID NO:575. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a NphB polypeptide, wherein the nucleotide sequence has at least 90% sequence identity to SEQ ID NO:575. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a NphB polypeptide, wherein the nucleotide sequence has at least 95% sequence identity to SEQ ID NO:575.

Polypeptides that Generate Acyl-CoA Compounds or Acyl-CoA Compound Derivatives [00710] A modified host cell of the present disclosure may comprise one or more heterologous nucleic acids comprising a nucleotide sequence encoding a polypeptide that generates acyl-CoA compounds or acyl-CoA compound derivatives. Such polypeptides may include, but are not limited to, acyl-activating enzyme (AAE) polypeptides, fatty acyl-CoA synthetases (FAA) polypeptides, or fatty acyl-CoA ligase polypeptides. In some

embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide.

[00711] AAE polypeptides, FAA polypeptides, and fatty acyl-CoA ligase

polypeptides can convert carboxylic acids to their CoA forms and generate acyl-CoA compounds or acyl-CoA compound derivatives. Promiscuous acyl-activating enzyme polypeptides, such as CsAAEl and CsAAE3 polypeptides, FAA polypeptides, or fatty acyl- CoA ligase polypeptides, may permit generation of cannabinoid derivatives (e.g., cannabigerolic acid derivatives), as well as cannabinoids (e.g., cannabigerolic acid). In some embodiments, unsubstituted or substituted hexanoic acid or carboxylic acids other than unsubstituted or substituted hexanoic acid are fed to modified host cells expressing an AAE polypeptide, FAA polypeptide, or fatty acyl-CoA ligase polypeptide (e.g., are present in the culture medium in which the cells are grown) to generate hexanoyl-CoA, acyl-CoA compounds, derivatives of hexanoyl-CoA, or derivatives of acyl-CoA compounds. The hexanoyl-CoA, acyl-CoA compounds, derivatives of hexanoyl-CoA, or derivatives of acyl- CoA compounds can then be further utilized by a modified host cell to generate

cannabinoids or cannabinoid derivatives. In certain such embodiments, the cell culture medium comprising the modified host cells comprises unsubstituted or substituted hexanoic acid. In some embodiments, the cell culture medium comprising the modified host cells comprises a carboxylic acid other than unsubstituted or substituted hexanoic acid.

[00712] Exemplary AAE, FAA, or fatty acyl-CoA ligase polypeptides disclosed herein may include a full-length AAE, FAA, or fatty acyl-CoA ligase polypeptide; a fragment of an AAE, FAA, or fatty acyl-CoA ligase polypeptide; a variant of an AAE, FAA, or fatty acyl-CoA ligase polypeptide; a truncated AAE, FAA, or fatty acyl-CoA ligase polypeptide; or a fusion polypeptide that has at least one activity of an AAE, FAA, or fatty acyl-CoA ligase polypeptide.

[00713] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the AAE polypeptide comprises the amino acid sequence set forth in SEQ ID NO:79, SEQ ID NO:268, or SEQ ID NO:270. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the AAE polypeptide comprises the amino acid sequence set forth in SEQ ID NO:79, SEQ ID NO:268, or SEQ ID NO:270, or a conservatively substituted amino acid sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the AAE polypeptide comprises an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, or at least 75% amino acid sequence identity to SEQ ID NO:79, SEQ ID NO:268, or SEQ ID NO:270. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the AAE polypeptide comprises an amino acid sequence having at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% amino acid sequence identity to SEQ ID NO:79, SEQ ID NO:268, or SEQ ID NO:270. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the AAE polypeptide comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:79, SEQ ID NO:268, or SEQ ID NO:270.

[00714] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the AAE polypeptide comprises the amino acid sequence set forth in SEQ ID NO:79 In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the AAE polypeptide comprises the amino acid sequence set forth in SEQ ID NO:79, or a conservatively substituted amino acid sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the AAE polypeptide comprises an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, or at least 75% amino acid sequence identity to SEQ ID NO:79. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the AAE polypeptide comprises an amino acid sequence having at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% amino acid sequence identity to SEQ ID NO:79. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the AAE polypeptide comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:79.

[00715] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the AAE polypeptide comprises the amino acid sequence set forth in SEQ ID NO:268. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the AAE polypeptide comprises the amino acid sequence set forth in SEQ ID NO:268, or a conservatively substituted amino acid sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the AAE polypeptide comprises an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, or at least 75% amino acid sequence identity to SEQ ID NO:268. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the AAE polypeptide comprises an amino acid sequence having at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% amino acid sequence identity to SEQ ID NO:268. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the AAE polypeptide comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:268.

[00716] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the AAE polypeptide comprises the amino acid sequence set forth in SEQ ID NO:270. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the AAE polypeptide comprises the amino acid sequence set forth in SEQ ID NO:270, or a conservatively substituted amino acid sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the AAE polypeptide comprises an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, or at least 75% amino acid sequence identity to SEQ ID NO:270. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the AAE polypeptide comprises an amino acid sequence having at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% amino acid sequence identity to SEQ ID NO:270. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the AAE polypeptide comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:270. In some embodiments, the AAE polypeptide comprises a C-terminal truncation.

[00717] Exemplary heterologous nucleic acids disclosed herein may include nucleic acids comprising a nucleotide sequence that encodes an AAE, FAA, or fatty acyl-CoA ligase polypeptide, such as, a full-length AAE, FAA, or fatty acyl-CoA ligase polypeptide; a fragment of an AAE, FAA, or fatty acyl-CoA ligase polypeptide; a variant of an AAE, FAA, or fatty acyl-CoA ligase polypeptide; a truncated AAE, FAA, or fatty acyl-CoA ligase polypeptide; or a fusion polypeptide that has at least one activity of an AAE, FAA, or fatty acyl-CoA ligase polypeptide. In some embodiments, the nucleotide sequence is codon- optimized.

[00718] In some embodiments, one or more AAE, FAA, or fatty acyl-CoA ligase polypeptide are overexpressed in the modified host cell. Overexpression may be achieved by increasing the copy number of the one or more heterologous nucleic acids comprising a nucleotide sequence encoding the AAE, FAA, or fatty acyl-CoA ligase polypeptide, e.g., through use of a high copy number expression vector (e.g., a plasmid that exists at 10-40 copies or about 100 copies per cell) and/or by operably linking the nucleotide sequence encoding the AAE, FAA, or fatty acyl-CoA ligase polypeptide to a strong promoter. In some embodiments, the modified host cell has one copy of a heterologous nucleic acid comprising a nucleotide sequence encoding an AAE, FAA, or fatty acyl-CoA ligase polypeptide. In some embodiments, the modified host cell has two copies of a heterologous nucleic acid comprising a nucleotide sequence encoding an AAE, FAA, or fatty acyl-CoA ligase polypeptide. In some embodiments, the modified host cell has three copies of a heterologous nucleic acid comprising a nucleotide sequence encoding an AAE, FAA, or fatty acyl-CoA ligase polypeptide. In some embodiments, the modified host cell has four copies of a heterologous nucleic acid comprising a nucleotide sequence encoding an AAE, FAA, or fatty acyl-CoA ligase polypeptide. In some embodiments, the modified host cell has five copies of a heterologous nucleic acid comprising a nucleotide sequence encoding an AAE, FAA, or fatty acyl-CoA ligase polypeptide. In some embodiments, the modified host cell has six copies of a heterologous nucleic acid comprising a nucleotide sequence encoding an AAE, FAA, or fatty acyl-CoA ligase polypeptide. In some embodiments, the modified host cell has seven copies of a heterologous nucleic acid comprising a nucleotide sequence encoding an AAE, FAA, or fatty acyl-CoA ligase polypeptide. In some embodiments, the modified host cell has eight copies of a heterologous nucleic acid comprising a nucleotide sequence encoding an AAE, FAA, or fatty acyl-CoA ligase polypeptide. In some embodiments, the modified host cell has eight or more copies of a heterologous nucleic acid comprising a nucleotide sequence encoding an AAE, FAA, or fatty acyl-CoA ligase polypeptide. Increased copy number of the heterologous nucleic acid and/or codon optimization of the nucleotide sequence may result in an increase in the desired enzyme catalytic activity in the modified host cell.

[00719] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:78, SEQ ID NO:266, SEQ ID NO:267, or SEQ ID NO:269. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:78, SEQ ID NO:266, SEQ ID NO:267, or SEQ ID N0 269, or a codon degenerate nucleotide sequence of any of the foregoing. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:78, SEQ ID NO:266, SEQ ID NO:267, or SEQ ID NO:269. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:78, SEQ ID NO:266, SEQ ID NO: 267, or SEQ ID NO: 269.

[00720] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:78. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:78, or a codon degenerate nucleotide sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:78. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:78.

[00721] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:266. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:266, or a codon degenerate nucleotide sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:266. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:266.

[00722] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:267. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:267, or a codon degenerate nucleotide sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:267. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:267.

[00723] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:269. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:269, or a codon degenerate nucleotide sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:269. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide, wherein the nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:269. In some

embodiments, the AAE polypeptide comprises a C-terminal truncation.

Polypeptides that Condense an Acyl-CoA Compound or an Acyl-CoA Compound Derivative with Malonyl-CoA to Generate Olivetolic Acid or Derivatives of Olivetolic Acid

[00724] A modified host cell of the present disclosure may comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides that condense an acyl-CoA compound, such as hexanoyl-CoA, or an acyl-CoA compound derivative, such as a hexanoyl-CoA derivative, with malonyl-CoA to generate olivetolic acid, or a derivative of olivetolic acid. Polypeptides that react an acyl-CoA compound or an acyl-CoA compound derivative with malonyl-CoA to generate olivetolic acid, or a derivative of olivetolic acid, may include TKS and OAC polypeptides. TKS and OAC polypeptides have been found to have broad substrate specificity, enabling production of cannabinoid derivatives or cannabinoids. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide.

[00725] Exemplary TKS or OAC polypeptides disclosed herein may include a full- length TKS or OAC polypeptide, a fragment of a TKS or OAC polypeptide, a variant of a TKS or OAC polypeptide, a truncated TKS or OAC polypeptide, or a fusion polypeptide that has at least one activity of a TKS or OAC polypeptide.

[00726] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide, wherein the TKS polypeptide comprises the amino acid sequence set forth in SEQ ID NO:75. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide, wherein the TKS polypeptide comprises the amino acid sequence set forth in SEQ ID NO:75, or a conservatively substituted amino acid sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide, wherein the TKS polypeptide comprises an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, or at least 75% amino acid sequence identity to SEQ ID NO:75. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide, wherein the TKS polypeptide comprises an amino acid sequence having at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% amino acid sequence identity to SEQ ID NO:75. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide, wherein the TKS polypeptide comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:75.

[00727] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide, wherein the OAC polypeptide comprises the amino acid sequence set forth in SEQ ID NO:77 or SEQ ID NO:225. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide, wherein the OAC polypeptide comprises the amino acid sequence set forth in SEQ ID NO:77 or SEQ ID NO:225, or a conservatively substituted amino acid sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide, wherein the OAC polypeptide comprises an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, or at least 75% amino acid sequence identity to SEQ ID NO:77 or SEQ ID NO:225. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide, wherein the OAC polypeptide comprises an amino acid sequence having at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% amino acid sequence identity to SEQ ID NO:77 or SEQ ID NO:225. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide, wherein the OAC polypeptide comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:77 or SEQ ID NO:225. [00728] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide, wherein the OAC polypeptide comprises the amino acid sequence set forth in SEQ ID NO:77. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide, wherein the OAC polypeptide comprises the amino acid sequence set forth in SEQ ID NO:77, or a conservatively substituted amino acid sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide, wherein the OAC polypeptide comprises an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, or at least 75% amino acid sequence identity to SEQ ID NO:77. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide, wherein the OAC polypeptide comprises an amino acid sequence having at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% amino acid sequence identity to SEQ ID NO:77. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide, wherein the OAC polypeptide comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:77.

[00729] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide, wherein the OAC polypeptide is a variant OAC (Y27F variant) polypeptide comprising the amino acid sequence set forth in SEQ ID NO:225. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide, wherein the OAC polypeptide is a variant OAC (Y27F variant) polypeptide comprising the amino acid sequence set forth in SEQ ID NO:225, or a conservatively substituted amino acid sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide, wherein the OAC polypeptide is a variant OAC (Y27F variant) polypeptide comprising an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, or at least 75% amino acid sequence identity to SEQ ID NO:225. In some

embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide, wherein the OAC polypeptide is a variant OAC (Y27F variant) polypeptide comprising an amino acid sequence having at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% amino acid sequence identity to SEQ ID NO:225. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide, wherein the OAC polypeptide is a variant OAC (Y27F variant) polypeptide comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:225.

[00730] Exemplary heterologous nucleic acids disclosed herein may include nucleic acids comprising a nucleotide sequence that encodes a TKS or OAC polypeptide, such as, a full-length TKS or OAC polypeptide, a fragment of a TKS or OAC polypeptide, a variant of a TKS or OAC polypeptide, a truncated TKS or OAC polypeptide, or a fusion polypeptide that has at least one activity of a TKS or OAC polypeptide. In some embodiments, the nucleotide sequence is codon-optimized.

[00731] In some embodiments, the TKS polypeptide is overexpressed in the modified host cell. Overexpression may be achieved by increasing the copy number of the one or more heterologous nucleic acids comprising a nucleotide sequence encoding the TKS polypeptide, e g., through use of a high copy number expression vector (e g., a plasmid that exists at 10-40 copies or about 100 copies per cell) and/or by operably linking the nucleotide sequence encoding the TKS polypeptide to a strong promoter. In some embodiments, the modified host cell has one copy of a heterologous nucleic acid comprising a nucleotide sequence encoding the TKS polypeptide. In some embodiments, the modified host cell has two copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the TKS polypeptide. In some embodiments, the modified host cell has three copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the TKS polypeptide. In some embodiments, the modified host cell has four copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the TKS polypeptide. In some embodiments, the modified host cell has five copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the TKS polypeptide. In some embodiments, the modified host cell has six copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the TKS polypeptide. In some embodiments, the modified host cell has seven copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the TKS polypeptide. In some embodiments, the modified host cell has eight copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the TKS polypeptide. In some embodiments, the modified host cell has nine copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the TKS polypeptide. In some embodiments, the modified host cell has ten copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the TKS polypeptide. In some embodiments, the modified host cell has eleven copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the TKS polypeptide. In some embodiments, the modified host cell has twelve copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the TKS polypeptide. In some

embodiments, the modified host cell has twelve or more copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the TKS polypeptide. Increased copy number of the heterologous nucleic acid and/or codon optimization of the nucleotide sequence may result in an increase in the desired enzyme catalytic activity in the modified host cell.

[00732] In some embodiments, the OAC polypeptide is overexpressed in the modified host cell. Overexpression may be achieved by increasing the copy number of the one or more heterologous nucleic acids comprising a nucleotide sequence encoding the OAC polypeptide, e.g., through use of a high copy number expression vector (e.g., a plasmid that exists at 10-40 copies or about 100 copies per cell) and/or by operably linking the nucleotide sequence encoding the OAC polypeptide to a strong promoter. In some embodiments, the modified host cell has one copy of a heterologous nucleic acid comprising a nucleotide sequence encoding the OAC polypeptide. In some embodiments, the modified host cell has two copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the OAC polypeptide. In some embodiments, the modified host cell has three copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the OAC polypeptide. In some embodiments, the modified host cell has four copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the OAC polypeptide. In some embodiments, the modified host cell has five copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the OAC polypeptide. In some embodiments, the modified host cell has six copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the OAC polypeptide. In some embodiments, the modified host cell has seven copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the OAC polypeptide. In some embodiments, the modified host cell has eight copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the OAC polypeptide. In some embodiments, the modified host cell has nine copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the OAC polypeptide. In some embodiments, the modified host cell has ten copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the OAC polypeptide. In some embodiments, the modified host cell has eleven copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the OAC polypeptide. In some embodiments, the modified host cell has twelve copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the OAC polypeptide. In some

embodiments, the modified host cell has twelve or more copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the OAC polypeptide. Increased copy number of the heterologous nucleic acid and/or codon optimization of the nucleotide sequence may result in an increase in the desired enzyme catalytic activity in the modified host cell.

[00733] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:74. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:74, or a codon degenerate nucleotide sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide, wherein the nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO: 74. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide, wherein the nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:74.

[00734] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:76 or SEQ ID NO: 224. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:76 or SEQ ID NO:224, or a codon degenerate nucleotide sequence of any of the foregoing. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide, wherein the nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:76 or SEQ ID NO:224. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide, wherein the nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:76 or SEQ ID NO:224.

[00735] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:76. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:76, or a codon degenerate nucleotide sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide, wherein the nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:76. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide, wherein the nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:76.

[00736] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide, wherein the OAC polypeptide is a variant OAC (Y27F variant) polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:224. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide, wherein the OAC polypeptide is a variant OAC (Y27F variant) polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:224, or a codon degenerate nucleotide sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide, wherein the OAC polypeptide is a variant OAC (Y27F variant) polypeptide, wherein the nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:224. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide, wherein the OAC polypeptide is a variant OAC (Y27F variant) polypeptide, wherein the nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:224.

Polypeptides that Generate Geranyl Pyrophosphate

[00737] A modified host cell of the present disclosure may comprise one or more heterologous nucleic acids comprising a nucleotide sequence encoding a polypeptide that generates GPP. In some embodiments, the polypeptide that generates GPP is a geranyl pyrophosphate synthetase (GPPS) polypeptide. In some embodiments, the GPPS polypeptide also has farnesyl diphosphate synthase (FPPS) polypeptide activity. In some embodiments, the GPPS polypeptide is modified such that it has reduced FPPS polypeptide activity (e g., at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or more than 90%, less FPPS polypeptide activity) than the corresponding wild-type or parental GPPS polypeptide from which the modified GPPS polypeptide is derived. In some embodiments, the GPPS polypeptide is modified such that it has substantially no FPPS polypeptide activity. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GPPS polypeptide.

[00738] Exemplary GPPS polypeptides disclosed herein may include a full-length GPPS polypeptide, a fragment of a GPPS polypeptide, a variant of a GPPS polypeptide, a truncated GPPS polypeptide, or a fusion polypeptide that has at least one activity of a GPPS polypeptide. [00739] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GPPS polypeptide, wherein the GPPS polypeptide is a variant GPPS (ERG20mut, F96W, N127W) polypeptide comprising the amino acid sequence set forth in SEQ ID NO:97. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GPPS polypeptide, wherein the GPPS polypeptide is a variant GPPS (ERG20mut, F96W, N127W) polypeptide comprising the amino acid sequence set forth in SEQ ID NO:97, or a conservatively substituted amino acid sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GPPS polypeptide, wherein the GPPS polypeptide is a variant GPPS

(ERG20mut, F96W, N127W) polypeptide comprising an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, or at least 75% amino acid sequence identity to SEQ ID NO:97. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GPPS polypeptide, wherein the GPPS polypeptide is a variant GPPS (ERG20mut, F96W, N127W) polypeptide comprising an amino acid sequence having at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% amino acid sequence identity to SEQ ID NO:97. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GPPS polypeptide, wherein the GPPS polypeptide is a variant GPPS

(ERG20mut, F96W, N127W) polypeptide comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:97. The mutation in this amino acid sequence shifts the ratio of GPP to farnesyl diphosphate (FPP), increasing the production of the GPP required to produce cannabinoids or cannabinoids derivatives.

[00740] Exemplary heterologous nucleic acids disclosed herein may include nucleic acids comprising a nucleotide sequence that encodes a GPPS polypeptide, such as, a full- length GPPS polypeptide, a fragment of a GPPS polypeptide, a variant of a GPPS polypeptide, a truncated GPPS polypeptide, or a fusion polypeptide that has at least one activity of a GPPS polypeptide. In some embodiments, the nucleotide sequence is codon- optimized. [00741] In some embodiments, the GPPS polypeptide is overexpressed in the modified host cell. Overexpression may be achieved by increasing the copy number of the one or more heterologous nucleic acids comprising a nucleotide sequence encoding the GPPS polypeptide, e.g., through use of a high copy number expression vector (e.g., a plasmid that exists at 10-40 copies or about 100 copies per cell) and/or by operably linking the nucleotide sequence encoding the GPPS polypeptide to a strong promoter. In some embodiments, the modified host cell has one copy of a heterologous nucleic acid comprising a nucleotide sequence encoding the GPPS polypeptide. In some embodiments, the modified host cell has two copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the GPPS polypeptide. In some embodiments, the modified host cell has three copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the GPPS polypeptide. In some embodiments, the modified host cell has four copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the GPPS polypeptide. In some embodiments, the modified host cell has five copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the GPPS polypeptide. In some embodiments, the modified host cell has six copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the GPPS polypeptide. In some embodiments, the modified host cell has seven copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the GPPS polypeptide. In some embodiments, the modified host cell has eight copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the GPPS polypeptide. In some embodiments, the modified host cell has eight or more copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the GPPS polypeptide. Increased copy number of the heterologous nucleic acid and/or codon optimization of the nucleotide sequence may result in an increase in the desired enzyme catalytic activity in the modified host cell.

[00742] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GPPS polypeptide, wherein the GPPS polypeptide is a variant GPPS (ERG20mut, F96W, N127W) polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:96. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GPPS polypeptide, wherein the GPPS polypeptide is a variant GPPS (ERG20mut, F96W, N127W) polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:96, or a codon degenerate nucleotide sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GPPS polypeptide, wherein the GPPS polypeptide is a variant GPPS (ERG20mut, F96W, N127W) polypeptide, wherein the nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO: 96. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GPPS polypeptide, wherein the GPPS polypeptide is a variant GPPS (ERG20mut, F96W, N127W) polypeptide, wherein the nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:96.

Polypeptides that Generate Acetyl-CoA from Pyruvate

[00743] A modified host cell of the present disclosure may comprise one or more heterologous nucleic acids comprising a nucleotide sequence encoding a polypeptide that generates acetyl-CoA from pyruvate. Polypeptides that generate acetyl-CoA from pyruvate may include a pyruvate decarboxylase (PDC) polypeptide. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a PDC polypeptide.

[00744] Exemplary PDC polypeptides disclosed herein may include a full-length PDC polypeptide, a fragment of a PDC polypeptide, a variant of a PDC polypeptide, a truncated PDC polypeptide, or a fusion polypeptide that has at least one activity of a PDC polypeptide.

[00745] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a PDC polypeptide, wherein the PDC polypeptide comprises the amino acid sequence set forth in SEQ ID NO:91. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a PDC polypeptide, wherein the PDC polypeptide comprises the amino acid sequence set forth in SEQ ID NO:9l, or a conservatively substituted amino acid sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a PDC polypeptide, wherein the PDC polypeptide comprises an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, or at least 75% amino acid sequence identity to SEQ ID NO:9l . In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a PDC polypeptide, wherein the PDC polypeptide comprises an amino acid sequence having at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% amino acid sequence identity to SEQ ID NO:9l. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a PDC polypeptide, wherein the PDC polypeptide comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:9l .

[00746] Exemplary heterologous nucleic acids disclosed herein may include nucleic acids comprising a nucleotide sequence that encodes a PDC polypeptide, such as, a full- length PDC polypeptide, a fragment of a PDC polypeptide, a variant of a PDC polypeptide, a truncated PDC polypeptide, or a fusion polypeptide that has at least one activity of a PDC polypeptide. In some embodiments, the nucleotide sequence is codon-optimized.

[00747] In some embodiments, the PDC polypeptide is overexpressed in the modified host cell. Overexpression may be achieved by increasing the copy number of the one or more heterologous nucleic acids comprising a nucleotide sequence encoding the PDC polypeptide, e.g., through use of a high copy number expression vector (e.g., a plasmid that exists at 10-40 copies or about 100 copies per cell) and/or by operably linking the nucleotide sequence encoding the PDC polypeptide to a strong promoter. In some embodiments, the modified host cell has one copy of a heterologous nucleic acid comprising a nucleotide sequence encoding the PDC polypeptide. In some embodiments, the modified host cell has two copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the PDC polypeptide. In some embodiments, the modified host cell has three copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the PDC polypeptide. In some embodiments, the modified host cell has four copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the PDC polypeptide. In some embodiments, the modified host cell has five copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the PDC polypeptide. In some embodiments, the modified host cell has five or more copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the PDC polypeptide. Increased copy number of the heterologous nucleic acid and/or codon optimization of the nucleotide sequence may result in an increase in the desired enzyme catalytic activity in the modified host cell. [00748] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a PDC polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:90. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a PDC polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:90, or a codon degenerate nucleotide sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a PDC polypeptide, wherein the nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO: 90. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a PDC polypeptide, wherein the nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:90.

Polypeptides that Condense Two Molecules of Acetyl-CoA to Generate Acetoacetyl-CoA [00749] A modified host cell of the disclosure may comprise one or more

heterologous nucleic acids comprising a nucleotide sequence encoding a polypeptide that condenses two molecules of acetyl-CoAto generate acetoacetyl-CoA. In some embodiments, the polypeptide that condenses two molecules of acetyl-CoA to generate acetoacetyl-CoA is an acetoacetyl-CoA thiolase polypeptide. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide.

[00750] Exemplary acetoacetyl-CoA thiolase polypeptides disclosed herein may include a full-length acetoacetyl-CoA thiolase polypeptide, a fragment of an acetoacetyl- CoA thiolase polypeptide, a variant of an acetoacetyl-CoA thiolase polypeptide, a truncated acetoacetyl-CoA thiolase polypeptide, or a fusion polypeptide that has at least one activity of an acetoacetyl-CoA thiolase polypeptide.

[00751] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl- CoA thiolase polypeptide, wherein the acetoacetyl-CoA thiolase polypeptide comprises the amino acid sequence set forth in SEQ ID NO:87. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide, wherein the acetoacetyl-CoA thiolase polypeptide comprises the amino acid sequence set forth in SEQ ID NO:87, or a conservatively substituted amino acid sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide, wherein the acetoacetyl-CoA thiolase polypeptide comprises an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, or at least 75% amino acid sequence identity to SEQ ID NO: 87. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide, wherein the acetoacetyl-CoA thiolase polypeptide comprises an amino acid sequence having at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% amino acid sequence identity to SEQ ID NO: 87. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide, wherein the acetoacetyl-CoA thiolase polypeptide comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:87.

[00752] Exemplary heterologous nucleic acids disclosed herein may include nucleic acids comprising a nucleotide sequence that encodes an acetoacetyl-CoA thiolase polypeptide, such as, a full-length acetoacetyl-CoA thiolase polypeptide, a fragment of an acetoacetyl-CoA thiolase polypeptide, a variant of an acetoacetyl-CoA thiolase polypeptide, a truncated acetoacetyl-CoA thiolase polypeptide, or a fusion polypeptide that has at least one activity of an acetoacetyl-CoA thiolase polypeptide. In some embodiments, the nucleotide sequence is codon-optimized.

[00753] In some embodiments, the acetoacetyl-CoA thiolase polypeptide is overexpressed in the modified host cell. Overexpression may be achieved by increasing the copy number of the one or more heterologous nucleic acids comprising a nucleotide sequence encoding the acetoacetyl-CoA thiolase polypeptide, e.g., through use of a high copy number expression vector (e.g., a plasmid that exists at 10-40 copies or about 100 copies per cell) and/or by operably linking the nucleotide sequence encoding the acetoacetyl- CoA thiolase polypeptide to a strong promoter. In some embodiments, the modified host cell has one copy of a heterologous nucleic acid comprising a nucleotide sequence encoding the acetoacetyl-CoA thiolase polypeptide. In some embodiments, the modified host cell has two copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the acetoacetyl-CoA thiolase polypeptide. In some embodiments, the modified host cell has three copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the acetoacetyl-CoA thiolase polypeptide. In some embodiments, the modified host cell has four copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the acetoacetyl-CoA thiolase polypeptide. In some embodiments, the modified host cell has five copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the acetoacetyl-CoA thiolase polypeptide. In some embodiments, the modified host cell has five or more copies of a heterologous nucleic acid comprising a nucleotide sequence encoding the acetoacetyl-CoA thiolase polypeptide. Increased copy number of the heterologous nucleic acid and/or codon optimization of the nucleotide sequence may result in an increase in the desired enzyme catalytic activity in the modified host cell.

[00754] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl- CoA thiolase polypeptide, wherein the nucleotide sequence is that set forth in SEQ ED NO: 86. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:86, or a codon degenerate nucleotide sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide, wherein the nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:86. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide, wherein the nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:86.

Mevalonate Pathway Polypeptides

[00755] A modified host cell of the present disclosure may comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides having at least one activity of a polypeptide present in the mevalonate (MEV) pathway. In certain such embodiments, the one or more polypeptides having at least one activity of a polypeptide present in the mevalonate (MEV) pathway comprise one or more MEV pathway polypeptides.

[00756] In some embodiments, the one or more polypeptides that are part of a biosynthetic pathway that generates GPP are one or more polypeptides having at least one activity of a polypeptide present in the mevalonate pathway. The mevalonate pathway may comprise polypeptides that catalyze the following steps: (a) condensing two molecules of acetyl-CoAto generate acetoacetyl-CoA (e.g., by action of an acetoacetyl-CoA thiolase polypeptide); (b) condensing acetoacetyl-CoA with acetyl-CoAto form

hydroxymethylglutaryl-CoA (HMG-CoA) (e.g., by action of a HMGS polypeptide); (c) converting HMG-CoA to mevalonate (e.g., by action of an HMGR polypeptide); (d) phosphorylating mevalonate to mevalonate 5-phosphate (e.g., by action of a MK

polypeptide); (e) converting mevalonate 5-phosphate to mevalonate 5-pyrophosphate (e.g., by action of a PMK polypeptide); (f) converting mevalonate 5 -pyrophosphate to isopentenyl pyrophosphate (e.g., by action of a mevalonate pyrophosphate decarboxylase (MPD or MVD1) polypeptide); and (g) converting isopentenyl pyrophosphate to dimethylallyl pyrophosphate (e.g., by action of an isopentenyl pyrophosphate isomerase (IDI1) polypeptide).

[00757] In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a MEV pathway polypeptide. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more MEV pathway polypeptide. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding two or more MEV pathway polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding three or more MEV pathway polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding four or more MEV pathway polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding five or more MEV pathway polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding six or more MEV pathway polypeptides. In some embodiments, a modified host cell of the present disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding all MEV pathway polypeptides.

[00758] Exemplary MEV pathway polypeptides disclosed herein may include a full- length MEV pathway polypeptide, a fragment of a MEV pathway polypeptide, a variant of a MEV pathway polypeptide, a truncated MEV pathway polypeptide, or a fusion polypeptide that has at least one activity of a MEV pathway polypeptide. In some embodiments, the one or more MEV pathway polypeptides are selected from the group consisting of an acetoacetyl-CoA thiolase polypeptide, a HMGS polypeptide, a HMGR polypeptide, an MK polypeptide, a PMK polypeptide, an MVD1 polypeptide, and an IDI1 polypeptide.

[00759] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMGS polypeptide, wherein the HMGS polypeptide comprises the amino acid sequence set forth in SEQ ID NO:85. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMGS polypeptide, wherein the HMGS polypeptide comprises the amino acid sequence set forth in SEQ ID NO:85, or a conservatively substituted amino acid sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMGS polypeptide, wherein the HMGS polypeptide comprises an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, or at least 75% amino acid sequence identity to SEQ ID NO:85. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMGS polypeptide, wherein the HMGS polypeptide comprises an amino acid sequence having at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% amino acid sequence identity to SEQ ID NO:85. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMGS polypeptide, wherein the HMGS polypeptide comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:85.

[00760] In some embodiments, the HMGR polypeptide is a truncated HMGR

(tHMGR) polypeptide. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a tHMGR polypeptide, wherein the tHMGR polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 83. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a tHMGR polypeptide, wherein the tHMGR polypeptide comprises the amino acid sequence set forth in SEQ ID NO:83, or a conservatively substituted amino acid sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a tHMGR polypeptide, wherein the tHMGR polypeptide comprises an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, or at least 75% amino acid sequence identity to SEQ ED NO: 83. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a tHMGR polypeptide, wherein the tHMGR polypeptide comprises an amino acid sequence having at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% amino acid sequence identity to SEQ ID NO: 83. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a tHMGR polypeptide, wherein the tHMGR polypeptide comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO: 83.

[00761] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a MK polypeptide, wherein the MK polypeptide comprises the amino acid sequence set forth in SEQ ID NO:95. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a MK polypeptide, wherein the MK polypeptide comprises the amino acid sequence set forth in SEQ ID NO:95, or a conservatively substituted amino acid sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a MK polypeptide, wherein the MK polypeptide comprises an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, or at least 75% amino acid sequence identity to SEQ ID NO:95. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a MK polypeptide, wherein the MK polypeptide comprises an amino acid sequence having at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% amino acid sequence identity to SEQ ID NO:95. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a MK polypeptide, wherein the MK polypeptide comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:95.

[00762] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a PMK polypeptide, wherein the PMK polypeptide comprises the amino acid sequence set forth in SEQ ID NO:93. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a PMK polypeptide, wherein the PMK polypeptide comprises the amino acid sequence set forth in SEQ ID NO:93, or a conservatively substituted amino acid sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a PMK polypeptide, wherein the PMK polypeptide comprises an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, or at least 75% amino acid sequence identity to SEQ ID NO:93. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a PMK polypeptide, wherein the PMK polypeptide comprises an amino acid sequence having at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% amino acid sequence identity to SEQ ID NO:93. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a PMK polypeptide, wherein the PMK polypeptide comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:93.

[00763] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a MVD1 polypeptide, wherein the MVD1 polypeptide comprises the amino acid sequence set forth in SEQ ID NO:89. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a MYD1 polypeptide, wherein the MVD1 polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 89, or a conservatively substituted amino acid sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a MVD1 polypeptide, wherein the MVD1 polypeptide comprises an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, or at least 75% amino acid sequence identity to SEQ ID NO:89. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a MVD1 polypeptide, wherein the MVD1 polypeptide comprises an amino acid sequence having at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% amino acid sequence identity to SEQ ID NO:89. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a MVD1 polypeptide, wherein the MVD1 polypeptide comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:89.

[00764] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an IDI1 polypeptide, wherein the IDI1 polypeptide comprises the amino acid sequence set forth in SEQ ID NO:81. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an IDI1 polypeptide, wherein the IDI1 polypeptide comprises the amino acid sequence set forth in SEQ ID NO:8l, or a conservatively substituted amino acid sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an IDI1 polypeptide, wherein the IDI1 polypeptide comprises an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, or at least 75% amino acid sequence identity to SEQ ID NO:8l . In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an IDI1 polypeptide, wherein the IDI1 polypeptide comprises an amino acid sequence having at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% amino acid sequence identity to SEQ ID NO:8l. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an IDI1 polypeptide, wherein the IDI1 polypeptide comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to SEQ ID NO:8l .

[00765] Exemplary heterologous nucleic acids disclosed herein may include nucleic acids comprising a nucleotide sequence that encodes a MEV pathway polypeptide, such as, a full-length MEV pathway polypeptide, a fragment of a MEV pathway polypeptide, a variant of a MEV pathway polypeptide, a truncated MEV pathway polypeptide, or a fusion polypeptide that has at least one activity of a polypeptide that is part of the MEV pathway.

In some embodiments, the nucleotide sequence is codon-optimized.

[00766] In some embodiments, one or more MEV pathway polypeptides are overexpressed in the modified host cell. Overexpression may be achieved by increasing the copy number of the one or more heterologous nucleic acids comprising a nucleotide sequence encoding a MEV pathway polypeptide, e.g., through use of a high copy number expression vector (e.g., a plasmid that exists at 10-40 copies or about 100 copies per cell) and/or by operably linking the nucleotide sequence encoding a MEV pathway polypeptide to a strong promoter. In some embodiments, the modified host cell has one copy of a heterologous nucleic acid comprising a nucleotide sequence encoding a MEV pathway polypeptide. In some embodiments, the modified host cell has two copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a MEV pathway polypeptide. In some embodiments, the modified host cell has three copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a MEV pathway polypeptide. In some embodiments, the modified host cell has four copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a MEV pathway polypeptide. In some embodiments, the modified host cell has five copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a MEV pathway polypeptide. In some embodiments, the modified host cell has five or more copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a MEV pathway polypeptide. Increased copy number of the heterologous nucleic acid and/or codon optimization of the nucleotide sequence may result in an increase in the desired enzyme catalytic activity in the modified host cell.

[00767] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMGS polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:84. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMGS polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO: 84, or a codon degenerate nucleotide sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMGS polypeptide, wherein the nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO: 84. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMGS polypeptide, wherein the nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:84.

[00768] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising nucleotide sequence encoding a tHMGR polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:82. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a tHMGR polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO: 82, or a codon degenerate nucleotide sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a tHMGR polypeptide, wherein the nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO: 82. In some

embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a tHMGR polypeptide, wherein the nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:82. [00769] In some embodiments, a modified host cell of the present disclosure comprises two or more heterologous nucleic acids comprising a nucleotide sequence that encodes a tHMGR polypeptide. In some embodiments, a modified host cell of the present disclosure comprises two heterologous nucleic acids comprising a nucleotide sequence that encodes a tHMGR polypeptide.

[00770] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a MK polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:94. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a MK polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:94, or a codon degenerate nucleotide sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a MK polypeptide, wherein the nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO: 94. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a MK polypeptide, wherein the nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:94.

[00771] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a PMK polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:92. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a PMK polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:92, or a codon degenerate nucleotide sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a PMK polypeptide, wherein the nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO: 92. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a PMK polypeptide, wherein the nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:92.

[00772] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a MVD1 polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:88. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a MVD1 polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:88, or a codon degenerate nucleotide sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a MVD1 polypeptide, wherein the nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO:88. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a MVDl polypeptide, wherein the nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:88.

[00773] In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an IDI1 polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO:80. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an IDI1 polypeptide, wherein the nucleotide sequence is that set forth in SEQ ID NO: 80, or a codon degenerate nucleotide sequence thereof. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an IDI1 polypeptide, wherein the nucleotide sequence has at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% sequence identity to SEQ ID NO: 80. In some embodiments, a modified host cell of the disclosure comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an IDI1 polypeptide, wherein the nucleotide sequence has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to SEQ ID NO:80.

Modified Host Cells to Generate Cannabinoids or Cannabinoid Derivatives and/or Express Cannabinoid Synthase Polypeptides

[00774] The disclosure provides for modified host cells for producing cannabinoids or cannabinoid derivatives. For producing cannabinoids or cannabinoid derivatives, modified host cells disclosed herein may be modified to express or overexpress one or more heterologous nucleic acids disclosed herein comprising nucleotide sequences encoding a cannabinoid synthase polypeptide, one or more secretory pathway polypeptides, and/or one or more polypeptides involved in cannabinoid or cannabinoid precursor (e.g.,

geranylpyrophosphate (GPP), prenyl phosphates, olivetolic acid, or hexanoyl-CoA) biosynthesis. In some embodiments, the nucleotide sequences encoding the one or more polypeptides involved in cannabinoid or cannabinoid precursor biosynthesis are codon- optimized. In some embodiments, the nucleotide sequences encoding the one or more secretory pathway polypeptides are codon-optimized. A modified host cell for producing cannabinoids or cannabinoid derivatives may comprise a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides. In certain such embodiments, the modified host cell for producing cannabinoids or cannabinoid derivatives may comprise a deletion of one or more genes encoding one or more secretory pathway polypeptides. In some embodiments, the modified host cell for producing cannabinoids or cannabinoid derivatives may comprise a downregulation of one or more genes encoding one or more secretory pathway polypeptides. In some embodiments, the nucleotide sequence encoding the cannabinoid synthase polypeptide is codon-optimized. In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide, such as a secretory signal sequence polypeptide. In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the cannabinoid synthase polypeptide is a fusion polypeptide with an AGA2t polypeptide. In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the cannabinoid synthase polypeptide is a fusion polypeptide with a GFP polypeptide. In the modified host cells for producing a cannabinoid or a cannabinoid derivative disclosed herein, the cannabinoid synthase polypeptide may include a tetrahydrocannabinolic acid synthase polypeptide, a cannabichromenic acid synthase polypeptide, or a cannabidiolic acid synthase polypeptide. [00775] The disclosure also provides modified host cells modified to express or overexpress one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide. In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and one or more heterologous nucleic acids disclosed herein comprising nucleotide sequences encoding secretory pathway polypeptides. In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides. In certain such embodiments, the modified host cell may comprise a deletion of one or more genes encoding one or more secretory pathway polypeptides. In some embodiments, the modified host cell may comprise a downregulation of one or more genes encoding one or more secretory pathway polypeptides. In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the nucleotide sequence encoding the cannabinoid synthase polypeptide comprises a codon-optimized nucleotide sequence. In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide, such as a secretory signal sequence polypeptide. In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the cannabinoid synthase polypeptide is a fusion polypeptide with an AGA2t polypeptide. In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the cannabinoid synthase polypeptide is a fusion polypeptide with a GFP polypeptide. In the modified host cells for expressing a cannabinoid synthase polypeptide disclosed herein, the cannabinoid synthase polypeptide may include a tetrahydrocannabinolic acid synthase polypeptide, a cannabichromenic acid synthase polypeptide, or a cannabidiolic acid synthase polypeptide. In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in

cannabinoid or cannabinoid precursor biosynthesis. In some embodiments, the nucleotide sequences encoding the one or more secretory pathway polypeptides and/or one or more polypeptides involved in cannabinoid or cannabinoid precursor biosynthesis are codon- optimized. [00776] To produce cannabinoids or cannabinoid derivatives, expression or overexpression of one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide in a modified host cell may be done in combination with expression or overexpression by the modified host cell of one or more other heterologous nucleic acids disclosed herein (e.g., one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides) and/or with deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides. In some embodiments, the nucleotide sequence encoding a cannabinoid synthase polypeptide comprises a codon-optimized nucleotide sequence. In some embodiments, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide, such as a secretory signal sequence polypeptide. In some embodiments, the cannabinoid synthase polypeptide is a fusion polypeptide with an AGA2t polypeptide. In some embodiments, the cannabinoid synthase polypeptide is a fusion polypeptide with a GFP polypeptide. In some embodiments, the nucleotide sequences encoding the one or more secretory pathway polypeptides are codon-optimized.

[00777] To express or overexpress cannabinoid synthase polypeptides, expression or overexpression of one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide in a modified host cell may be done in combination with expression or overexpression by the modified host cell of one or more other heterologous nucleic acids disclosed herein (e g., one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides) and/or with deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides In some embodiments, the nucleotide sequence encoding a cannabinoid synthase polypeptide comprises a codon-optimized nucleotide sequence. In some embodiments, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide, such as a secretory signal sequence polypeptide. In some embodiments, the cannabinoid synthase polypeptide is a fusion polypeptide with an AGA2t polypeptide. In some embodiments, the cannabinoid synthase polypeptide is a fusion polypeptide with a GFP polypeptide. In some embodiments, the nucleotide sequences encoding the one or more secretory pathway polypeptides are codon-optimized.

[00778] In some embodiments, the growth and/or viability of modified host cells of the disclosure for producing cannabinoids or cannabinoid derivatives is not significantly decreased compared to the growth and/or viability of an unmodified host cell. In some embodiments, a culture of modified host cells of the disclosure for producing cannabinoids or cannabinoid derivatives has a cell density that is at least 25% or greater, at least 30% or greater, at least 35% or greater, at least 40% or greater, at least 45% or greater, at least 50% or greater, at least 55% or greater, at least 60% or greater, at least 65% or greater, at least 70% or greater, at least 75% or greater, at least 80% or greater, at least 85% or greater at least 90% or greater, at least 95% or greater, at least 100% or greater, at least 110% or greater, at least 120% or greater, at least 130% or greater, at least 140% or greater, or at least 150% or greater than the cell density of a culture of unmodified control host cells grown for the same period, in the same culture medium, and under the same culture conditions.

[00779] In some embodiments, the growth and/or viability of modified host cells of the disclosure for expressing a cannabinoid synthase polypeptide is not significantly decreased compared to the growth and/or viability of an unmodified host cell. In some embodiments, a culture of modified host cells of the disclosure for expressing a cannabinoid synthase polypeptide has a cell density that is at least 25% or greater, at least 30% or greater, at least 35% or greater, at least 40% or greater, at least 45% or greater, at least 50% or greater, at least 55% or greater, at least 60% or greater, at least 65% or greater, at least 70% or greater, at least 75% or greater, at least 80% or greater, at least 85% or greater at least 90% or greater, at least 95% or greater, at least 100% or greater, at least 110% or greater, at least 120% or greater, at least 130% or greater, at least 140% or greater, or at least 150% or greater than the cell density of a culture of unmodified control host cells grown for the same period, in the same culture medium, and under the same culture conditions.

[00780] In some embodiments, the growth and/or viability of modified host cells of the disclosure comprising one or more heterologous nucleic acids comprising a codon- optimized nucleotide sequence encoding a cannabinoid synthase polypeptide is not significantly decreased compared to the growth and/or viability of an unmodified host cell.

In some embodiments, a culture of modified host cells of the disclosure comprising one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide has a cell density that is at least 25% or greater, at least 30% or greater, at least 35% or greater, at least 40% or greater, at least 45% or greater, at least 50% or greater, at least 55% or greater, at least 60% or greater, at least 65% or greater, at least 70% or greater, at least 75% or greater, at least 80% or greater, at least 85% or greater at least 90% or greater, at least 95% or greater, at least 100% or greater, at least 110% or greater, at least 120% or greater, at least 130% or greater, at least 140% or greater, or at least 150% or greater than the cell density of a culture of unmodified control host cells grown for the same period, in the same culture medium, and under the same culture conditions. In some embodiments of the modified host cells of the disclosure comprising one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide, such as a secretory signal sequence polypeptide. In some embodiments of the modified host cells of the disclosure comprising one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, the cannabinoid synthase polypeptide is a fusion polypeptide with an AGA2t polypeptide. In some embodiments of the modified host cells of the disclosure comprising one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, the cannabinoid synthase polypeptide is a fusion polypeptide with a GFP polypeptide. In some embodiments of the modified host cells of the disclosure comprising one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide, the modified host cells comprise one or more

heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cannabinoid or cannabinoid precursor (e.g., geranylpyrophosphate (GPP), prenyl phosphates, olivetolic acid, or hexanoyl-CoA) biosynthesis. In some embodiments, the nucleotide sequences encoding the one or more polypeptides involved in cannabinoid or cannabinoid precursor (e g., geranylpyrophosphate (GPP), prenyl phosphates, olivetolic acid, or hexanoyl-CoA) biosynthesis are codon-optimized.

[00781] In some embodiments, the growth and/or viability of modified host cells of the disclosure comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and one or more modifications to modulate the expression of one or more secretory pathway polypeptides is not significantly decreased compared to the growth and/or viability of an unmodified host cell. In some embodiments, a culture of modified host cells of the disclosure comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and one or more modifications to modulate the expression of one or more secretory pathway polypeptides has a cell density that is at least 25% or greater, at least 30% or greater, at least 35% or greater, at least 40% or greater, at least 45% or greater, at least 50% or greater, at least 55% or greater, at least 60% or greater, at least 65% or greater, at least 70% or greater, at least 75% or greater, at least 80% or greater, at least 85% or greater at least 90% or greater, at least 95% or greater, at least 100% or greater, at least 110% or greater, at least 120% or greater, at least 130% or greater, at least 140% or greater, or at least 150% or greater than the cell density of a culture of unmodified control host cells grown for the same period, in the same culture medium, and under the same culture conditions. In certain such embodiments, the nucleotide sequence encoding the cannabinoid synthase polypeptide is codon-optimized. In some embodiments of the modified host cells of the disclosure comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and one or more modifications to modulate the expression of one or more secretory pathway polypeptides, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide, such as a secretory signal sequence polypeptide. In some embodiments of the modified host cells of the disclosure comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and one or more modifications to modulate the expression of one or more secretory pathway polypeptides, the cannabinoid synthase polypeptide is a fusion polypeptide with an AGA2t polypeptide. In some embodiments of the modified host cells of the disclosure comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and one or more modifications to modulate the expression of one or more secretory pathway polypeptides, the cannabinoid synthase polypeptide is a fusion polypeptide with a GFP polypeptide. In some embodiments of the modified host cells of the disclosure comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and one or more modifications to modulate the expression of one or more secretory pathway polypeptides, the modified host cells comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cannabinoid or cannabinoid precursor (e g., geranylpyrophosphate (GPP), prenyl phosphates, olivetolic acid, or hexanoyl-CoA) biosynthesis. In some embodiments, the nucleotide sequences encoding the one or more secretory pathway polypeptides, and/or one or more polypeptides involved in cannabinoid or cannabinoid precursor (e.g., geranylpyrophosphate (GPP), prenyl phosphates, olivetolic acid, or hexanoyl-CoA) biosynthesis are codon-optimized.

[00782] In some embodiments, the growth and/or viability of modified host cells of the disclosure comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides is not significantly decreased compared to the growth and/or viability of an unmodified host cell. In some embodiments, a culture of modified host cells of the disclosure comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides has a cell density that is at least 25% or greater, at least 30% or greater, at least 35% or greater, at least 40% or greater, at least 45% or greater, at least 50% or greater, at least 55% or greater, at least 60% or greater, at least 65% or greater, at least 70% or greater, at least 75% or greater, at least 80% or greater, at least 85% or greater at least 90% or greater, at least 95% or greater, at least 100% or greater, at least 110% or greater, at least 120% or greater, at least 130% or greater, at least 140% or greater, or at least 150% or greater than the cell density of a culture of unmodified control host cells grown for the same period, in the same culture medium, and under the same culture conditions. In certain such embodiments, the nucleotide sequence encoding the cannabinoid synthase polypeptide is codon-optimized. In some embodiments of the modified host cells of the disclosure comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide, such as a secretory signal sequence polypeptide. In some embodiments of the modified host cells of the disclosure comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, the cannabinoid synthase polypeptide is a fusion polypeptide with an AGA2t polypeptide. In some embodiments of the modified host cells of the disclosure comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, the cannabinoid synthase polypeptide is a fusion polypeptide with a GFP polypeptide. In some embodiments of the modified host cells of the disclosure comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, the modified host cells comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cannabinoid or cannabinoid precursor (e.g., geranylpyrophosphate (GPP), prenyl phosphates, olivetolic acid, or hexanoyl-CoA) biosynthesis. In some embodiments, the nucleotide sequences encoding the one or more secretory pathway polypeptides, and/or one or more polypeptides involved in cannabinoid or cannabinoid precursor (e.g., geranylpyrophosphate (GPP), prenyl phosphates, olivetolic acid, or hexanoyl-CoA) biosynthesis are codon-optimized.

[00783] In some embodiments, the growth and/or viability of modified host cells of the disclosure comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides is not significantly decreased compared to the growth and/or viability of an unmodified host cell. In some embodiments, a culture of modified host cells of the disclosure comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides has a cell density that is at least 25% or greater, at least 30% or greater, at least 35% or greater, at least 40% or greater, at least 45% or greater, at least 50% or greater, at least 55% or greater, at least 60% or greater, at least 65% or greater, at least 70% or greater, at least 75% or greater, at least 80% or greater, at least 85% or greater at least 90% or greater, at least 95% or greater, at least 100% or greater, at least 110% or greater, at least 120% or greater, at least 130% or greater, at least 140% or greater, or at least 150% or greater than the cell density of a culture of unmodified control host cells grown for the same period, in the same culture medium, and under the same culture conditions. In certain such embodiments, the nucleotide sequence encoding the cannabinoid synthase polypeptide is codon-optimized. In some embodiments of the modified host cells of the disclosure comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide, such as a secretory signal sequence polypeptide. In some embodiments of the modified host cells of the disclosure comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, the cannabinoid synthase polypeptide is a fusion polypeptide with an AGA2t polypeptide. In some embodiments of the modified host cells of the disclosure comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, the cannabinoid synthase polypeptide is a fusion polypeptide with a GFP polypeptide. In some embodiments of the modified host cells of the disclosure comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, the modified host cells comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cannabinoid or cannabinoid precursor (e.g., geranylpyrophosphate (GPP), prenyl phosphates, olivetolic acid, or hexanoyl-CoA) biosynthesis. In some embodiments, the nucleotide sequences encoding the one or more polypeptides involved in cannabinoid or cannabinoid precursor (e g., geranylpyrophosphate (GPP), prenyl phosphates, olivetolic acid, or hexanoyl-CoA) biosynthesis are codon-optimized.

[00784] In some embodiments, the growth and/or viability of modified host cells of the disclosure comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides, and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides is not significantly decreased compared to the growth and/or viability of an unmodified host cell. In some embodiments, a culture of modified host cells of the disclosure comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides, and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides has a cell density that is at least 25% or greater, at least 30% or greater, at least 35% or greater, at least 40% or greater, at least 45% or greater, at least 50% or greater, at least 55% or greater, at least 60% or greater, at least 65% or greater, at least 70% or greater, at least 75% or greater, at least 80% or greater, at least 85% or greater at least 90% or greater, at least 95% or greater, at least 100% or greater, at least 110% or greater, at least 120% or greater, at least 130% or greater, at least 140% or greater, or at least 150% or greater than the cell density of a culture of unmodified control host cells grown for the same period, in the same culture medium, and under the same culture conditions. In certain such embodiments, the nucleotide sequence encoding the cannabinoid synthase polypeptide is codon-optimized. In some embodiments of the modified host cells of the disclosure comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides, and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide, such as a secretory signal sequence polypeptide. In some

embodiments of the modified host cells of the disclosure comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, the cannabinoid synthase polypeptide is a fusion polypeptide with an AGA2t polypeptide. In some embodiments of the modified host cells of the disclosure comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, the cannabinoid synthase polypeptide is a fusion polypeptide with a GFP polypeptide. In some embodiments of the modified host cells of the disclosure comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, the modified host cells comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cannabinoid or cannabinoid precursor (e.g., geranylpyrophosphate (GPP), prenyl phosphates, olivetolic acid, or hexanoyl-CoA) biosynthesis. In some embodiments, the nucleotide sequences encoding the one or more secretory pathway polypeptides, and/or one or more polypeptides involved in cannabinoid or cannabinoid precursor (e.g., geranylpyrophosphate (GPP), prenyl phosphates, olivetolic acid, or hexanoyl-CoA) biosynthesis are codon-optimized.

Exemplary Modified Host Cells for Producing Cannabinoids or Cannabinoid Derivatives [00785] The present disclosure provides a modified host cell for producing cannabinoids or cannabinoid derivatives, wherein the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide. In certain such embodiments, the nucleotide sequence encoding the cannabinoid synthase polypeptide is codon-optimized. In the modified host cells for producing a cannabinoid or a cannabinoid derivative disclosed herein, the cannabinoid synthase polypeptide may include a tetrahydrocannabinolic acid synthase polypeptide, a cannabichromenic acid synthase polypeptide, or a cannabidiolic acid synthase polypeptide.

In some embodiments disclosed herein, the nucleotide sequences encoding the one or more secretory pathway polypeptides, and/or one or more polypeptides involved in cannabinoid or cannabinoid precursor (e.g., geranylpyrophosphate (GPP), prenyl phosphates, olivetolic acid, or hexanoyl-CoA) biosynthesis are codon-optimized.

[00786] The present disclosure provides a modified host cell for producing a cannabinoid or a cannabinoid derivative, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide and one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide; and c) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide.

[00787] The present disclosure provides a modified host cell for producing a cannabinoid or a cannabinoid derivative, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide.

[00788] The present disclosure provides a modified host cell for producing a cannabinoid or a cannabinoid derivative, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide; c) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMG-CoA synthase (HMGS) polypeptide; d) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3 -hydroxy-3 -methyl-glutaryl-CoA reductase (tHMGR) polypeptide; e) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate kinase (MK) polypeptide; f) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a phosphomevalonate kinase (PMK) polypeptide; g) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate pyrophosphate decarboxylase (MVD1) polypeptide; h) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a isopentenyl diphosphate isomerase (IDI1) polypeptide; i) one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide; j) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a pyruvate decarboxylase (PDC) polypeptide; and k) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate synthetase (GPPS) polypeptide.

[00789] The present disclosure provides a modified host cell for producing a cannabinoid or a cannabinoid derivative, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide; c) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMG-CoA synthase (HMGS) polypeptide; d) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3 -hydroxy-3 -methyl-glutaryl-CoA reductase (tHMGR) polypeptide; e) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate kinase (MK) polypeptide; f) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a phosphomevalonate kinase (PMK) polypeptide; g) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate pyrophosphate decarboxylase (MVD1) polypeptide; h) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a isopentenyl diphosphate isomerase (IDI1) polypeptide; i) one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide; and j) one or more

heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate synthetase (GPPS) polypeptide

[00790] The present disclosure provides a modified host cell for producing a cannabinoid or a cannabinoid derivative, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide; c) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMG-CoA synthase (HMGS) polypeptide; d) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3 -hydroxy-3 -methyl-glutaryl-CoA reductase (tHMGR) polypeptide; e) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate kinase (MK) polypeptide; f) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a phosphomevalonate kinase (PMK) polypeptide; g) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate pyrophosphate decarboxylase (MVD1) polypeptide; h) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a isopentenyl diphosphate isomerase (IDI1) polypeptide; i) one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide; j) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a pyruvate decarboxylase (PDC) polypeptide; k) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate synthetase (GPPS) polypeptide; and 1) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide and one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide.

[00791] The present disclosure provides a modified host cell for producing a cannabinoid or a cannabinoid derivative, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide; c) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMG-CoA synthase (HMGS) polypeptide; d) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3 -hydroxy-3 -methyl-glutaryl-CoA reductase (tHMGR) polypeptide; e) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate kinase (MK) polypeptide; f) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a phosphomevalonate kinase (PMK) polypeptide; g) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate pyrophosphate decarboxylase (MVD1) polypeptide; h) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a isopentenyl diphosphate isomerase (IDI1) polypeptide; i) one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide; j) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate synthetase (GPPS) polypeptide; and k) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide and one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide.

[00792] The present disclosure provides a modified host cell for producing a cannabinoid or a cannabinoid derivative, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide; c) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMG-CoA synthase (HMGS) polypeptide; d) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3 -hydroxy-3 -methyl-glutaryl-CoA reductase (tHMGR) polypeptide; e) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate kinase (MK) polypeptide; f) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a phosphomevalonate kinase (PMK) polypeptide; g) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate pyrophosphate decarboxylase (MVD1) polypeptide; h) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a isopentenyl diphosphate isomerase (IDI1) polypeptide; i) one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide; j) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a pyruvate decarboxylase (PDC) polypeptide; k) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate synthetase (GPPS) polypeptide; 1) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide and one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide; and m) one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acyl-activating enzyme (AAE) polypeptide.

[00793] The present disclosure provides a modified host cell for producing a cannabinoid or a cannabinoid derivative, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide; c) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMG-CoA synthase (HMGS) polypeptide; d) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3 -hydroxy-3 -methyl-glutaryl-CoA reductase (tHMGR) polypeptide; e) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate kinase (MK) polypeptide; f) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a phosphomevalonate kinase (PMK) polypeptide; g) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate pyrophosphate decarboxylase (MVD1) polypeptide; h) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a isopentenyl diphosphate isomerase (IDI1) polypeptide; i) one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide; j) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate synthetase (GPPS) polypeptide; k) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide and one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide; and 1) one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acyl-activating enzyme (AAE) polypeptide.

[00794] The present disclosure provides a modified host cell for producing a cannabinoid or a cannabinoid derivative, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide and one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide; c) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide; and d) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides. In certain such embodiments, the modified host cell for producing a cannabinoid or a cannabinoid derivative comprises a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides.

[00795] The present disclosure provides a modified host cell for producing a cannabinoid or a cannabinoid derivative, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide; and c) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides. In certain such embodiments, the modified host cell for producing a cannabinoid or a cannabinoid derivative comprises a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides.

[00796] The present disclosure provides a modified host cell for producing a cannabinoid or a cannabinoid derivative, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and b) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides. In certain such embodiments, the modified host cell for producing a cannabinoid or a cannabinoid derivative comprises a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides.

[00797] The present disclosure provides a modified host cell for producing a cannabinoid or a cannabinoid derivative, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide; c) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMG-CoA synthase (HMGS) polypeptide; d) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3 -hydroxy-3 -methyl-glutaryl-CoA reductase (tHMGR) polypeptide; e) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate kinase (MK) polypeptide; f) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a phosphomevalonate kinase (PMK) polypeptide; g) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate pyrophosphate decarboxylase (MVD1) polypeptide; h) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a isopentenyl diphosphate isomerase (IDI1) polypeptide; i) one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide; j) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a pyruvate decarboxylase (PDC) polypeptide; k) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate synthetase (GPPS) polypeptide; and 1) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides. In certain such embodiments, the modified host cell for producing a cannabinoid or a cannabinoid derivative comprises a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides.

[00798] The present disclosure provides a modified host cell for producing a cannabinoid or a cannabinoid derivative, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide; c) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMG-CoA synthase (HMGS) polypeptide; d) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3 -hydroxy-3 -methyl-glutaryl-CoA reductase (tHMGR) polypeptide; e) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate kinase (MK) polypeptide; f) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a phosphomevalonate kinase (PMK) polypeptide; g) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate pyrophosphate decarboxylase (MVD1) polypeptide; h) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a isopentenyl diphosphate isomerase (IDI1) polypeptide; i) one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide; j) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate synthetase (GPPS) polypeptide; and k) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides. In certain such embodiments, the modified host cell for producing a cannabinoid or a cannabinoid derivative comprises a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides.

[00799] The present disclosure provides a modified host cell for producing a cannabinoid or a cannabinoid derivative, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide; c) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMG-CoA synthase (HMGS) polypeptide; d) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3 -hydroxy-3 -methyl-glutaryl-CoA reductase (tHMGR) polypeptide; e) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate kinase (MK) polypeptide; f) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a phosphomevalonate kinase (PMK) polypeptide; g) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate pyrophosphate decarboxylase (MVD1) polypeptide; h) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a isopentenyl diphosphate isomerase (IDI1) polypeptide; i) one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide; j) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a pyruvate decarboxylase (PDC) polypeptide; k) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate synthetase (GPPS) polypeptide; 1) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide and one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide; and m) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides. In certain such embodiments, the modified host cell for producing a cannabinoid or a cannabinoid derivative comprises a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides.

[00800] The present disclosure provides a modified host cell for producing a cannabinoid or a cannabinoid derivative, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide; c) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMG-CoA synthase (HMGS) polypeptide; d) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3 -hydroxy-3 -methyl-glutaryl-CoA reductase (tHMGR) polypeptide; e) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate kinase (MK) polypeptide; f) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a phosphomevalonate kinase (PMK) polypeptide; g) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate pyrophosphate decarboxylase (MVD1) polypeptide; h) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a isopentenyl diphosphate isomerase (IDI1) polypeptide; i) one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide; j) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate synthetase (GPPS) polypeptide; k) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide and one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide; and 1) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides. In certain such embodiments, the modified host cell for producing a cannabinoid or a cannabinoid derivative comprises a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides.

[00801] The present disclosure provides a modified host cell for producing a cannabinoid or a cannabinoid derivative, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide; c) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMG-CoA synthase (HMGS) polypeptide; d) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3 -hydroxy-3 -methyl-glutaryl-CoA reductase (tHMGR) polypeptide; e) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate kinase (MK) polypeptide; f) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a phosphomevalonate kinase (PMK) polypeptide; g) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate pyrophosphate decarboxylase (MVD1) polypeptide; h) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a isopentenyl diphosphate isomerase (IDI1) polypeptide; i) one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide; j) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a pyruvate decarboxylase (PDC) polypeptide; k) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate synthetase (GPPS) polypeptide; 1) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide and one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide; m) one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acyl-activating enzyme (AAE) polypeptide; and n) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides. In certain such embodiments, the modified host cell for producing a cannabinoid or a cannabinoid derivative comprises a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides.

[00802] The present disclosure provides a modified host cell for producing a cannabinoid or a cannabinoid derivative, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide; c) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMG-CoA synthase (HMGS) polypeptide; d) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3 -hydroxy-3 -methyl-glutaryl-CoA reductase (tHMGR) polypeptide; e) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate kinase (MK) polypeptide; f) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a phosphomevalonate kinase (PMK) polypeptide; g) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate pyrophosphate decarboxylase (MVD1) polypeptide; h) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a isopentenyl diphosphate isomerase (IDI1) polypeptide; i) one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide; j) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate synthetase (GPPS) polypeptide; k) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide and one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide; 1) one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acyl-activating enzyme (AAE) polypeptide; and m) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides. In certain such embodiments, the modified host cell for producing a cannabinoid or a cannabinoid derivative comprises a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides.

[00803] The present disclosure provides a modified host cell for producing a cannabinoid or a cannabinoid derivative, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide and one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide; c) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide; and d) a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides. In certain such embodiments, the modified host cell for producing a cannabinoid or a cannabinoid derivative comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides.

[00804] The present disclosure provides a modified host cell for producing a cannabinoid or a cannabinoid derivative, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide; and c) a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides. In certain such embodiments, the modified host cell for producing a cannabinoid or a cannabinoid derivative comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides. [00805] The present disclosure provides a modified host cell for producing a cannabinoid or a cannabinoid derivative, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and b) a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides. In certain such embodiments, the modified host cell for producing a cannabinoid or a cannabinoid derivative comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides.

[00806] The present disclosure provides a modified host cell for producing a cannabinoid or a cannabinoid derivative, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide; c) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMG-CoA synthase (HMGS) polypeptide; d) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3 -hydroxy-3 -methyl-glutaryl-CoA reductase (tHMGR) polypeptide; e) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate kinase (MK) polypeptide; f) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a phosphomevalonate kinase (PMK) polypeptide; g) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate pyrophosphate decarboxylase (MVD1) polypeptide; h) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a isopentenyl diphosphate isomerase (IDI1) polypeptide; i) one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide; j) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a pyruvate decarboxylase (PDC) polypeptide; k) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate synthetase (GPPS) polypeptide; and 1) a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides. In certain such embodiments, the modified host cell for producing a cannabinoid or a cannabinoid derivative comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides.

[00807] The present disclosure provides a modified host cell for producing a cannabinoid or a cannabinoid derivative, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide; c) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMG-CoA synthase (HMGS) polypeptide; d) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3 -hydroxy-3 -methyl-glutaryl-CoA reductase (tHMGR) polypeptide; e) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate kinase (MK) polypeptide; f) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a phosphomevalonate kinase (PMK) polypeptide; g) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate pyrophosphate decarboxylase (MVD1) polypeptide; h) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a isopentenyl diphosphate isomerase (IDI1) polypeptide; i) one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide; j) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate synthetase (GPPS) polypeptide; and k) a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides. In certain such embodiments, the modified host cell for producing a cannabinoid or a cannabinoid derivative comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides.

[00808] The present disclosure provides a modified host cell for producing a cannabinoid or a cannabinoid derivative, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide; c) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMG-CoA synthase (HMGS) polypeptide; d) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3 -hydroxy-3 -methyl-glutaryl-CoA reductase (tHMGR) polypeptide; e) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate kinase (MK) polypeptide; f) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a phosphomevalonate kinase (PMK) polypeptide; g) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate pyrophosphate decarboxylase (MVD1) polypeptide; h) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a isopentenyl diphosphate isomerase (IDI1) polypeptide; i) one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide; j) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a pyruvate decarboxylase (PDC) polypeptide; k) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate synthetase (GPPS) polypeptide; 1) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide and one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide; and m) a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides. In certain such embodiments, the modified host cell for producing a cannabinoid or a cannabinoid derivative comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides.

[00809] The present disclosure provides a modified host cell for producing a cannabinoid or a cannabinoid derivative, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide; c) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMG-CoA synthase (HMGS) polypeptide; d) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3 -hydroxy-3 -methyl-glutaryl-CoA reductase (tHMGR) polypeptide; e) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate kinase (MK) polypeptide; f) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a phosphomevalonate kinase (PMK) polypeptide; g) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate pyrophosphate decarboxylase (MVD1) polypeptide; h) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a isopentenyl diphosphate isomerase (IDI1) polypeptide; i) one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide; j) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate synthetase (GPPS) polypeptide; k) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide and one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide; and 1) a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides. In certain such embodiments, the modified host cell for producing a cannabinoid or a cannabinoid derivative comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides.

[00810] The present disclosure provides a modified host cell for producing a cannabinoid or a cannabinoid derivative, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide; c) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMG-CoA synthase (HMGS) polypeptide; d) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3 -hydroxy-3 -methyl-glutaryl-CoA reductase (tHMGR) polypeptide; e) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate kinase (MK) polypeptide; f) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a phosphomevalonate kinase (PMK) polypeptide; g) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate pyrophosphate decarboxylase (MVD1) polypeptide; h) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a isopentenyl diphosphate isomerase (IDI1) polypeptide; i) one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide; j) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a pyruvate decarboxylase (PDC) polypeptide; k) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate synthetase (GPPS) polypeptide; 1) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide and one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide; m) one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acyl-activating enzyme (AAE) polypeptide; and n) a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides. In certain such embodiments, the modified host cell for producing a cannabinoid or a cannabinoid derivative comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides.

[00811] The present disclosure provides a modified host cell for producing a cannabinoid or a cannabinoid derivative, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide; c) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMG-CoA synthase (HMGS) polypeptide; d) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3 -hydroxy-3 -methyl-glutaryl-CoA reductase (tHMGR) polypeptide; e) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate kinase (MK) polypeptide; f) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a phosphomevalonate kinase (PMK) polypeptide; g) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate pyrophosphate decarboxylase (MVD1) polypeptide; h) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a isopentenyl diphosphate isomerase (IDI1) polypeptide; i) one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide; j) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate synthetase (GPPS) polypeptide; k) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide and one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide; 1) one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acyl-activating enzyme (AAE) polypeptide; and m) a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides. In certain such embodiments, the modified host cell for producing a cannabinoid or a cannabinoid derivative comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides.

[00812] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides. In certain such embodiments, the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin mononucleotide (FMN) synthetase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosidase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosyltransferase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more peptidyl-prolyl isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more fatty acid desaturase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein transport and trafficking polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more signal peptidase or signal peptidase complex polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR), one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in endoplasmic reticulum-associated degradation (ERAD), one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cell wall assembly, or one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in vacuolar protein sorting, or a combination of any of the foregoing.

[00813] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a JEM1 polypeptide and a KAR2 polypeptide. In some embodiments, the one or more chaperone or co-chaperone

polypeptides comprise a LHSl polypeptide and a KAR2 polypeptide. In some embodiments, the one or more chaperone or co-chaperone polypeptides comprise a SIL1 polypeptide and a KAR2 polypeptide. In some embodiments, the one or more chaperone or co-chaperone polypeptides comprise a CNE1 polypeptide and a KAR2 polypeptide. In some embodiments, the one or more chaperone or co-chaperone polypeptides comprise a SIS1 polypeptide and a KAR2 polypeptide. In some embodiments, the one or more chaperone or co-chaperone polypeptides comprise a SSB1 polypeptide and a KAR2 polypeptide. In some embodiments, the one or more chaperone or co-chaperone polypeptides comprise a ROT1 polypeptide and a KAR2 polypeptide. In some embodiments, the one or more chaperone or co-chaperone polypeptides comprise a DED1 polypeptide and a KAR2 polypeptide. In some

embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide. In some embodiments, the one or more chaperone or co-chaperone

polypeptides comprise a SSB1 polypeptide. In some embodiments, the one or more chaperone or co-chaperone polypeptides comprise a CPR5 polypeptide. In some

embodiments, the one or more chaperone or co-chaperone polypeptides comprise a LHS1 polypeptide. In some embodiments, the one or more chaperone or co-chaperone

polypeptides comprise a SSA1 polypeptide. In some embodiments, the one or more chaperone or co-chaperone polypeptides comprise a CNS1 polypeptide. In some embodiments, the one or more chaperone or co-chaperone polypeptides comprise a DED1 polypeptide. In some embodiments, the one or more chaperone or co-chaperone

polypeptides comprise a PFD2s polypeptide. In some embodiments, the one or more chaperone or co-chaperone polypeptides comprise a JEM1 polypeptide. In some

embodiments, the one or more chaperone or co-chaperone polypeptides comprise a CNE1 polypeptide. In some embodiments, the one or more chaperone or co-chaperone polypeptides comprise a SCJ1 polypeptide. In some embodiments, the one or more chaperone or co chaperone polypeptides comprise a SIL1 polypeptide. In some embodiments, the one or more chaperone or co-chaperone polypeptides comprise a FPRl polypeptide. In some embodiments, the one or more chaperone or co-chaperone polypeptides comprise a ROT1 polypeptide.

[00814] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum. In certain such embodiments, the one or more polypeptides involved in protein translocation into the endoplasmic reticulum comprise a SEC61 polypeptide, a SBH1 polypeptide, and a SSS1 polypeptide. In some embodiments, the one or more polypeptides involved in protein translocation into the endoplasmic reticulum comprise a SRP14 polypeptide, a SRP21 polypeptide, a SRP68 polypeptide, a SRP72 polypeptide, and a SRP54 polypeptide. In certain such embodiments, the modified host cell for producing a cannabinoid or a cannabinoid derivative comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding SRP protein complex RNA component SCR1. In some embodiments, the one or more polypeptides involved in protein translocation into the endoplasmic reticulum comprise a SRP101 polypeptide and a SRP102 polypeptide.

[00815] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR). In certain such embodiments, the one or more chaperone or co chaperone polypeptides comprise KAR2 and the one or more polypeptides involved in UPR comprise a HACls polypeptide.

[00816] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR). In certain such embodiments, the one or more chaperone or co chaperone polypeptides comprise a KAR2 polypeptide and the one or more polypeptides involved in UPR comprise an IRE1 polypeptide.

[00817] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR), one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides. In certain such embodiments, the one or more chaperone or co chaperone polypeptides comprise a KAR2 polypeptide, the one or more polypeptides involved in UPR comprise an IRE1 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, and the one or more thiol oxidase polypeptides comprise an EROl polypeptide.

[00818] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide and a DED1 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, and the one or more thiol oxidase polypeptides comprise an EROl polypeptide.

[00819] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR), one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides. In certain such embodiments, the one or more chaperone or co chaperone polypeptides comprise a KAR2 polypeptide, the one or more polypeptides involved in UPR comprise a PPQ1 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, and the one or more thiol oxidase polypeptides comprise an EROl polypeptide.

[00820] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein transport and trafficking polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide and the one or more protein transport and trafficking polypeptides comprise a BFR2 polypeptide.

[00821] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR) In certain such embodiments, the one or more polypeptides involved in UPR comprise a HACls polypeptide and/or an IRE1 polypeptide. In some embodiments, the one or more polypeptides involved in UPR comprise a HACls polypeptide. In some embodiments, the one or more polypeptides involved in unfolded protein response (UPR) comprise an IRE1 polypeptide.

[00822] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, and the one or more thiol oxidase polypeptides comprise an EROl polypeptide.

[00823] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, and the one or more thiol oxidase polypeptides comprise an ERV2 polypeptide.

[00824] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more thiol oxidase polypeptides comprise an EROl polypeptide, and the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide.

[00825] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in UPR, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide and a SSB1 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more thiol oxidase polypeptides comprise an EROl polypeptide, the one or more polypeptides involved in EPR comprise a HACls polypeptide, and the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide. In certain such embodiments, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide, wherein the signal sequence polypeptide is a vacuolar localization signal sequence polypeptide, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide or a PRClt polypeptide.

[00826] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in ERAD, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more polypeptides involved in ERAD comprise a NPL4 polypeptide, and the one or more thiol oxidase polypeptides comprise an EROl polypeptide.

[00827] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cell wall assembly, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more polypeptides involved in cell wall assembly comprise a KRE1 polypeptide, and the one or more thiol oxidase polypeptides comprise an EROl polypeptide.

[00828] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in vacuolar protein sorting, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more polypeptides involved in vacuolar protein sorting comprise a PEP1 polypeptide, and the one or more thiol oxidase polypeptides comprise an EROl polypeptide.

[00829] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein transport and trafficking polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more protein transport and trafficking polypeptides comprise a LDB17 polypeptide, and the one or more thiol oxidase polypeptides comprise an EROl polypeptide.

[00830] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides. In certain such embodiments, the one or more chaperone or co chaperone polypeptides comprise a KAR2 polypeptide, a PFD1 polypeptide, and a PFD2s polypeptide; the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide; the one or more polypeptides involved in protein translocation into the endoplasmic reticulum comprise a SRP14 polypeptide, a SRP21 polypeptide, a SRP68 polypeptide, a SRP72 polypeptide, a SEC65 polypeptide, and a SRP54 polypeptide; and the one or more thiol oxidase polypeptides comprise an EROl polypeptide. In certain such embodiments, the modified host cell for producing a cannabinoid or a cannabinoid derivative comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding SRP protein complex RNA component SCR1.

[00831] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides. In certain such embodiments, the one or more chaperone or co chaperone polypeptides comprise a KAR2 polypeptide; the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide; the one or more polypeptides involved in protein translocation into the endoplasmic reticulum comprise a SRP14 polypeptide, a SRP21 polypeptide, a SRP68 polypeptide, a SRP72 polypeptide, a SEC65 polypeptide, a SRP54 polypeptide, a SEC61 polypeptide, a SBH1 polypeptide, and a SSS1 polypeptide, and the one or more thiol oxidase polypeptides comprise an EROl polypeptide. In certain such embodiments, the modified host cell for producing a cannabinoid or a cannabinoid derivative comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding SRP protein complex RNA component SCR1.

[00832] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides. In certain such embodiments, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide and the one or more thiol oxidase polypeptides comprise an EROl polypeptide and an ERV2 polypeptide.

[00833] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR). In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide and a SSB1 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more thiol oxidase polypeptides comprise an EROl polypeptide, and the one or more polypeptides involved in EPR comprise a HACls polypeptide.

[00834] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides. In certain such embodiments, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide and the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide.

[00835] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin mononucleotide (FMN) synthetase polypeptides and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides. In certain such embodiments, the one or more flavin

mononucleotide (FMN) synthetase polypeptides comprise a FMN1 polypeptide and the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide.

[00836] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosidase polypeptides. In certain such embodiments, the one or more glycosidase polypeptides comprise a CWH41 polypeptide.

[00837] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cell wall assembly. In certain such embodiments, the one or more polypeptides involved in cell wall assembly comprise a KRE1 polypeptide.

[00838] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in ERAD. In certain such embodiments, the one or more polypeptides involved in ERAD comprise a NPL4 polypeptide. [00839] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, and the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide.

[00840] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide and the one or more protein disulfide isomerase polypeptides comprise a PDIl polypeptide.

[00841] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein transport and trafficking polypeptides. In certain such embodiments, the one or more protein transport and trafficking polypeptides comprise a SEC7 polypeptide. In some embodiments, the one or more protein transport and trafficking polypeptides comprise a BFR2 polypeptide. In some embodiments, the one or more protein transport and trafficking polypeptides comprise a SEC23 polypeptide, a SEC24 polypeptide, a SEC13 polypeptide, a SEC31 polypeptide, a SAR1 polypeptide, a SEC 12 polypeptide, a SEC4 polypeptide, a SEC 16 polypeptide, and an ERV29 polypeptide. In some embodiments, the one or more protein transport and trafficking polypeptides comprise a SEC5 polypeptide, a SEC22 polypeptide, a BOS1 polypeptide, a BETl polypeptide, a BE!Gl polypeptide, a GRH1 polypeptide, a USOl polypeptide, a SEC 17 polypeptide, a SEC 18 polypeptide, a SSOl polypeptide, a SEC9 polypeptide, and a SNC2 polypeptide.

[00842] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more signal peptidase or signal peptidase complex polypeptides. In certain such embodiments, the one or more signal peptidase or signal peptidase complex polypeptides comprise a KEX2 polypeptide. In some embodiments, the one or more signal peptidase or signal peptidase complex polypeptides comprise a SPC1 polypeptide, a SPC2 polypeptide, a SPC3 polypeptide, and a SEC11 polypeptide.

[00843] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein transport and trafficking polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide and the one or more protein transport and trafficking polypeptides comprise a SEC24 polypeptide.

[00844] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides. In certain such embodiments, the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more glycosidase polypeptides, one or more genes encoding one or more glycosyltransferase polypeptides, one or more genes encoding one or more polypeptides involved in lipid droplet assembly, one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism, or one or more genes encoding one or more vacuolar proteinase polypeptides, or a combination of any of the foregoing. In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises a deletion of one or more genes encoding one or more secretory pathway polypeptides. In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises a downregulation of one or more genes encoding one or more secretory pathway polypeptides.

[00845] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more glycosidase polypeptides and one or more genes encoding one or more polypeptides involved in lipid droplet assembly. In certain such embodiments, the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene and the one or more genes encoding one or more polypeptides involved in lipid droplet assembly comprise a FLD1 gene.

[00846] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more polypeptides involved in lipid droplet assembly. In certain such embodiments, the one or more genes encoding one or more polypeptides involved in lipid droplet assembly comprise a FLD1 gene.

[00847] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more glycosidase polypeptides. In certain such embodiments, the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 and/or MNS1 gene. In some embodiments, the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene. In some

embodiments, the one or more genes encoding one or more glycosidase polypeptides comprise a MNS1 gene.

[00848] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more glycosyltransferase polypeptides. In certain such embodiments, the one or more genes encoding one or more glycosyltransferase polypeptides comprise an ALG12 gene.

[00849] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides. In certain such embodiments, the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene.

[00850] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides. In certain such embodiments, the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin mononucleotide (FMN) synthetase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosidase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosyltransferase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more peptidyl-prolyl isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more fatty acid desaturase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein transport and trafficking polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more signal peptidase or signal peptidase complex polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR), one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in endoplasmic reticulum-associated degradation (ERAD), one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cell wall assembly, or one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in vacuolar protein sorting, or a combination of any of the foregoing, and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more glycosidase polypeptides, one or more genes encoding one or more glycosyltransferase polypeptides, one or more genes encoding one or more polypeptides involved in lipid droplet assembly, one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism, or one or more genes encoding one or more vacuolar proteinase polypeptides, or a combination of any of the foregoing. In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and a deletion of one or more genes encoding one or more secretory pathway polypeptides. In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and a downregulation of one or more genes encoding one or more secretory pathway polypeptides.

[00851] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more polypeptides involved in lipid droplet assembly. In certain such embodiments, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide, and the one or more genes encoding one or more polypeptides involved in lipid droplet assembly comprise a FLDl gene.

[00852] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more glycosidase polypeptides. In certain such embodiments, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide, and the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene.

[00853] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more glycosidase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide, and the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene.

[00854] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more glycosidase polypeptides and one or more genes encoding one or more vacuolar proteinase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene, and the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene.

[00855] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR), and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more glycosidase polypeptides and one or more genes encoding one or more vacuolar proteinase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more polypeptides involved in unfolded protein response (UPR) comprise an IRE1 polypeptide, the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene, and the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene.

[00856] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more glycosidase polypeptides and one or more genes encoding one or more polypeptides involved in lipid droplet assembly. In certain such embodiments, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide, the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene, and the one or more genes encoding one or more polypeptides involved in lipid droplet assembly comprise a FLD1 gene. [00857] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more glycosidase polypeptides, one or more genes encoding one or more polypeptides involved in lipid droplet assembly, and one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism. In certain such embodiments, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FADl polypeptide, the one or more thiol oxidase polypeptides comprise an EROl polypeptide, the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene, the one or more genes encoding one or more polypeptides involved in lipid droplet assembly comprise a FLD1 gene, and the one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism comprise an OPI1 gene

[00858] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides and one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a CPR5 polypeptide and a KAR2 polypeptide, the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide, the one or more thiol oxidase polypeptides comprise an EROl polypeptide, the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene, and the one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism comprise an OPI1 gene. In certain such embodiments, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide, wherein the signal sequence polypeptide is a vacuolar localization signal sequence polypeptide, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide.

[00859] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a CPR5 polypeptide and the one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism comprise an OPI1 gene. In certain such embodiments, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide, wherein the signal sequence polypeptide is a vacuolar localization signal sequence polypeptide, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide.

[00860] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides, one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism, and one or more genes encoding one or more glycosidase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a CPR5 polypeptide and a KAR2 polypeptide, the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide, the one or more thiol oxidase polypeptides comprise an EROl polypeptide, the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene, the one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism comprise an OPI1 gene, and the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene. In certain such embodiments, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide, wherein the signal sequence polypeptide is a vacuolar localization signal sequence polypeptide, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide.

[00861] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR), and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides, one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism, and one or more genes encoding one or more glycosidase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a CPR5 polypeptide and a KAR2 polypeptide, the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide, the one or more thiol oxidase polypeptides comprise an EROl polypeptide, the one or more polypeptides involved in unfolded protein response (UPR) comprise an IRE1 polypeptide, the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene, the one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism comprise an OPI1 gene, and the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene. In certain such embodiments, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide, wherein the signal sequence polypeptide is a vacuolar localization signal sequence polypeptide, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide.

[00862] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR), and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides, one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism, and one or more genes encoding one or more glycosidase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a CPR5 polypeptide, a DED1 polypeptide, and a KAR2 polypeptide, the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide, the one or more thiol oxidase polypeptides comprise an EROl polypeptide, the one or more polypeptides involved in unfolded protein response (UPR) comprise an IRE1 polypeptide, the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene, the one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism comprise an OPI1 gene, and the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene. In certain such embodiments, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide, wherein the signal sequence polypeptide is a vacuolar localization signal sequence polypeptide, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide.

[00863] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR), and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides and one or more genes encoding one or more glycosidase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide and a DED1 polypeptide, the one or more thiol oxidase polypeptides comprise an EROl polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more polypeptides involved in unfolded protein response (UPR) comprise an IRE1 polypeptide, the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene, and the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene. [00864] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR), and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides, one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism, and one or more genes encoding one or more glycosidase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a CPR5 polypeptide and a KAR2 polypeptide, the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide, the one or more thiol oxidase polypeptides comprise an EROl polypeptide, the one or more polypeptides involved in unfolded protein response (UPR) comprise an IRE1 polypeptide and a PPQ1 polypeptide, the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene, the one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism comprise an OPI1 gene, and the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene. In certain such embodiments, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide, wherein the signal sequence polypeptide is a vacuolar localization signal sequence polypeptide, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide.

[00865] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in vacuolar protein sorting, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR), and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides, one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism, and one or more genes encoding one or more glycosidase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a CPR5 polypeptide and a KAR2 polypeptide, the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide, the one or more thiol oxidase polypeptides comprise an EROl polypeptide, the one or more polypeptides involved in vacuolar protein sorting comprise a PEP1 polypeptide, the one or more polypeptides involved in unfolded protein response (LJPR) comprise an IRE1 polypeptide, the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene, the one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism comprise an OPI1 gene, and the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene. In certain such embodiments, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide, wherein the signal sequence polypeptide is a vacuolar localization signal sequence polypeptide, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide.

[00866] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in vacuolar protein sorting, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR), and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides and one or more genes encoding one or more glycosidase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide, the one or more thiol oxidase polypeptides comprise an EROl polypeptide, the one or more polypeptides involved in vacuolar protein sorting comprise a PEP1 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more polypeptides involved in unfolded protein response (UPR) comprise an IRE1 polypeptide, the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene, and the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene.

[00867] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein transport and trafficking polypeptides, and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides and one or more genes encoding one or more glycosidase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more protein transport and trafficking polypeptides comprise a SEC24 polypeptide, the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene, and the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene.

[00868] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR), and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides and one or more genes encoding one or more glycosidase polypeptides. In certain such embodiments, the one or more chaperone or co chaperone polypeptides comprise a KAR2 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more thiol oxidase polypeptides comprise an EROl polypeptide, the one or more polypeptides involved in unfolded protein response (UPR) comprise an IRE1 polypeptide, the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene, and the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene.

[00869] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR), and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides and one or more genes encoding one or more glycosidase polypeptides. In certain such embodiments, the one or more chaperone or co chaperone polypeptides comprise a KAR2 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more thiol oxidase polypeptides comprise an ERV2 polypeptide, the one or more polypeptides involved in unfolded protein response (UPR) comprise an IRE1 polypeptide, the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene, and the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene.

[00870] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more thiol oxidase polypeptides comprise an EROl polypeptide, and the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene.

[00871] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, and the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene.

[00872] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein transport and trafficking polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR), and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides, one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism, and one or more genes encoding one or more glycosidase polypeptides. In certain such embodiments, the one or more chaperone or co chaperone polypeptides comprise a CPR5 polypeptide and a KAR2 polypeptide, the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide, the one or more thiol oxidase polypeptides comprise an EROl polypeptide, the one or more protein transport and trafficking polypeptides comprise a SEC24 polypeptide, the one or more polypeptides involved in unfolded protein response (UPR) comprise an IRE1 polypeptide, the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene, the one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism comprise an OPI1 gene, and the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene. In certain such embodiments, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide, wherein the signal sequence polypeptide is a vacuolar localization signal sequence polypeptide, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide.

[00873] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR), and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides, one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism, and one or more genes encoding one or more glycosidase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a CPR5 polypeptide and a KAR2 polypeptide, the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide, the one or more thiol oxidase polypeptides comprise an EROl polypeptide and an ERV2 polypeptide, the one or more polypeptides involved in unfolded protein response (UPR) comprise an IRE1 polypeptide, the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene, the one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism comprise an OPI1 gene, and the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene. In certain such embodiments, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide, wherein the signal sequence polypeptide is a vacuolar localization signal sequence polypeptide, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide.

[00874] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises:

a) two or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide;

1) two or more heterologous nucleic acids comprising a nucleotide sequence encoding a thiol oxidase polypeptide, wherein the thiol oxidase polypeptide is an EROl polypeptide;

In certain such embodiments, the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide, one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide, and/or one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acyl-activating enzyme (AAE) polypeptide. In some embodiments, the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00875] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises:

a) two heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide;

[00876] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises:

i) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a pyruvate decarboxylase (PDC) polypeptide; j) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate synthetase (GPPS) polypeptide;

[00877] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises: a) two heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide;

1) two heterologous nucleic acids comprising a nucleotide sequence encoding a chaperone or co-chaperone polypeptide, wherein the chaperone or co- chaperone polypeptide is a KAR2 polypeptide;

2) a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in unfolded protein response, wherein the polypeptide involved in unfolded protein response is an IRE1 polypeptide; and 3) a heterologous nucleic acid comprising a nucleotide sequence encoding a protein disulfide isomerase polypeptide, wherein the protein disulfide isomerase polypeptide is a PDIl polypeptide; and

[00878] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises:

g) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a isopentenyl diphosphate isomerase (IDI1) polypeptide; h) one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide;

[00879] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises: a) two heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide;

c) two heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3-hydroxy-3-methyl-glutaryl-CoA reductase (tHMGR) polypeptide; d) a heterologous nucleic acid comprising a nucleotide sequence encoding a mevalonate kinase (MK) polypeptide;

1) a heterologous nucleic acid comprising a nucleotide sequence encoding a chaperone or co-chaperone polypeptide, wherein the chaperone or co chaperone polypeptide is a KAR2 polypeptide; and

2) a heterologous nucleic acid comprising a nucleotide sequence encoding a protein disulfide isomerase polypeptide, wherein the protein disulfide isomerase polypeptide is a PDIl polypeptide; and m) deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise:

[00880] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises:

a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide;

[00881] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises:

a) a heterologous nucleic acid comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide;

b) a heterologous nucleic acid comprising a nucleotide sequence encoding a HMG- CoA synthase (HMGS) polypeptide; c) two heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3-hydroxy-3-methyl-glutaryl-CoA reductase (tHMGR) polypeptide; d) a heterologous nucleic acid comprising a nucleotide sequence encoding a mevalonate kinase (MK) polypeptide;

k) a heterologous nucleic acid comprising a nucleotide sequence encoding a geranyl pyrophosphate:olivetolic acid geranyltransferase (GOT) polypeptide; and l) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise:

3) a heterologous nucleic acid comprising a nucleotide sequence encoding a flavin adenine dinucleotide (FAD) synthetase polypeptide, wherein the flavin adenine dinucleotide (FAD) synthetase polypeptide is a FADl polypeptide; and 4) a heterologous nucleic acid comprising a nucleotide sequence encoding a protein disulfide isomerase polypeptide, wherein the protein disulfide isomerase polypeptide is a PDIl polypeptide.

[00882] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises:

j) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate synthetase (GPPS) polypeptide; k) two or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate:olivetolic acid geranyltransferase (GOT) polypeptide; and

[00883] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises:

k) two heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate:olivetolic acid geranyltransferase (GOT) polypeptide; and l) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise:

4) a heterologous nucleic acid comprising a nucleotide sequence encoding a protein disulfide isomerase polypeptide, wherein the protein disulfide isomerase polypeptide is a PDIl polypeptide. In certain such embodiments, the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide, one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide, and/or one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acyl-activating enzyme (AAE) polypeptide. In some embodiments, the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00884] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises:

5) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a chaperone or co-chaperone polypeptide, wherein the chaperone or co-chaperone polypeptide is a SSB1 polypeptide; and

6) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a polypeptide involved in unfolded protein response, wherein the polypeptide involved in unfolded protein response is a HACls polypeptide.

[00885] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises:

4) a heterologous nucleic acid comprising a nucleotide sequence encoding a protein disulfide isomerase polypeptide, wherein the protein disulfide isomerase polypeptide is a PDIl polypeptide; 5) a heterologous nucleic acid comprising a nucleotide sequence encoding a chaperone or co-chaperone polypeptide, wherein the chaperone or co chaperone polypeptide is a SSB1 polypeptide; and

[00886] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises:

[00887] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises:

e) a heterologous nucleic acid comprising a nucleotide sequence encoding a phosphomevalonate kinase (PMK) polypeptide; f) a heterologous nucleic acid comprising a nucleotide sequence encoding a mevalonate pyrophosphate decarboxylase (MVD1) polypeptide;

[00888] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises two or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide. In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises three or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide. In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises three or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide. In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises two or more heterologous nucleic acids comprising a nucleotide sequence encoding an acyl-activating enzyme (AAE) polypeptide. In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises two or more heterologous nucleic acids comprising a nucleotide sequence encoding a tHMGR polypeptide. In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises two or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide.

[00889] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises two heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide. In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises three heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide. In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises three heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide. In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises two heterologous nucleic acids comprising a nucleotide sequence encoding an acyl-activating enzyme (AAE) polypeptide. In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises two heterologous nucleic acids comprising a nucleotide sequence encoding a tHMGR polypeptide. In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises two heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide.

[00890] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises a) two or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; b) three or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide; c) three or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide; and d) two or more heterologous nucleic acids comprising a nucleotide sequence encoding an acyl-activating enzyme (AAE) polypeptide. In certain such embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises two or more heterologous nucleic acids comprising a nucleotide sequence encoding a tHMGR

polypeptide.

[00891] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises a) two heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; b) three heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide; c) three heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide; and d) two heterologous nucleic acids comprising a nucleotide sequence encoding an acyl-activating enzyme (AAE) polypeptide. In certain such embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises two heterologous nucleic acids comprising a nucleotide sequence encoding a tHMGR polypeptide.

[00892] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises a) three or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide; b) three or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide; and c) two or more heterologous nucleic acids comprising a nucleotide sequence encoding an acyl-activating enzyme (AAE) polypeptide. In certain such embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises two or more heterologous nucleic acids comprising a nucleotide sequence encoding a tHMGR polypeptide.

[00893] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises a) three heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide; b) three heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide; and c) two heterologous nucleic acids comprising a nucleotide sequence encoding an acyl-activating enzyme (AAE) polypeptide. In certain such embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises two heterologous nucleic acids comprising a nucleotide sequence encoding a tHMGR

polypeptide. [00894] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises two or more heterologous nucleic acids comprising a nucleotide sequence encoding a KAR2 polypeptide. In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises three or more heterologous nucleic acids comprising a nucleotide sequence encoding a KAR2 polypeptide. In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises four or more heterologous nucleic acids comprising a nucleotide sequence encoding a KAR2 polypeptide. In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises two or more heterologous nucleic acids comprising a nucleotide sequence encoding an EROl polypeptide. In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises two or more heterologous nucleic acids comprising a nucleotide sequence encoding a PDI1 polypeptide.

[00895] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises two heterologous nucleic acids comprising a nucleotide sequence encoding a KAR2 polypeptide. In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises three heterologous nucleic acids comprising a nucleotide sequence encoding a KAR2 polypeptide. In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises four

heterologous nucleic acids comprising a nucleotide sequence encoding a KAR2 polypeptide. In some embodiments of the modified host cell for producing a cannabinoid or a

cannabinoid derivative, the modified host cell comprises two heterologous nucleic acids comprising a nucleotide sequence encoding an EROl polypeptide. In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the modified host cell comprises two heterologous nucleic acids comprising a nucleotide sequence encoding a PDI1 polypeptide.

[00896] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the nucleotide sequence encoding the cannabinoid synthase polypeptide is codon-optimized.

[00897] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide. In certain such embodiments, the one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide comprise a nucleotide sequence encoding a signal sequence polypeptide. In some embodiments, the signal sequence polypeptide is a secretory signal sequence polypeptide. In some embodiments, the secretory signal sequence polypeptide is a native secretory signal sequence polypeptide. In some embodiments, the secretory signal sequence polypeptide is a synthetic secretory signal sequence polypeptide. In some embodiments, the secretory signal sequence polypeptide is an endoplasmic reticulum retention signal sequence polypeptide. In certain such embodiments, the endoplasmic reticulum retention signal sequence polypeptide is a HDEL polypeptide or a KDEL polypeptide. In some embodiments, the secretory signal sequence polypeptide is a mitochondrial targeting signal sequence polypeptide. In some embodiments, the secretory signal sequence polypeptide is a Golgi targeting signal sequence polypeptide. In some embodiments, the secretory signal sequence polypeptide is a vacuolar localization signal sequence polypeptide. In certain such embodiments, the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide or a PRClt polypeptide. In certain such embodiments, the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide. In some embodiments, the secretory signal sequence polypeptide is a plasma membrane localization signal sequence polypeptide. In some embodiments, the secretory signal sequence polypeptide is a peroxisome targeting signal sequence polypeptide. In some embodiments, the peroxisome targeting signal sequence polypeptide is a PEX8 polypeptide. In some embodiments, the secretory signal sequence polypeptide is a mating factor secretory signal sequence polypeptide (e.g., a MF polypeptide or an evolved MF polypeptide (MFev)). In some embodiments, the signal sequence polypeptide is linked to the N-terminus of the cannabinoid synthase polypeptide.

[00898] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the cannabinoid synthase polypeptide is a fusion polypeptide with an AGA2t polypeptide. In certain such embodiments, the one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide comprise a nucleotide sequence encoding an AGA2t polypeptide. In certain such embodiments, the modified host cell for producing a cannabinoid or a cannabinoid derivative is modified with one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AGA1 polypeptide. In certain such embodiments, the cannabinoid synthase-AGA2t fusion polypeptide may be displayed on the cell surface of the modified host cell. In some embodiments, the AGA2t polypeptide is linked to the N-terminus of the cannabinoid synthase polypeptide. [00899] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the cannabinoid synthase polypeptide is a fusion polypeptide with a GFP polypeptide. In certain such embodiments, the one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide comprise a nucleotide sequence encoding a GFP polypeptide. In some embodiments, the GFP polypeptide is linked to the N-terminus of the cannabinoid synthase polypeptide.

Exemplary Modified Host Cells for Expressing Cannabinoid Synthase Polypeptides

[00900] The present disclosure provides a modified host cell for expressing a cannabinoid synthase polypeptide, wherein the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide. In certain such embodiments, the nucleotide sequence encoding a cannabinoid synthase polypeptide is codon-optimized. In the modified host cells for expressing cannabinoid synthases disclosed herein, the cannabinoid synthase polypeptide may include a tetrahydrocannabinolic acid synthase polypeptide, a cannabichromenic acid synthase polypeptide, or a cannabidiolic acid synthase polypeptide. In some embodiments disclosed herein, the nucleotide sequences encoding the one or more secretory pathway polypeptides, and/or one or more polypeptides involved in cannabinoid or cannabinoid precursor (e.g., geranylpyrophosphate (GPP), prenyl phosphates, olivetolic acid, or hexanoyl-CoA) biosynthesis are codon-optimized.

[00901] The present disclosure provides a modified host cell for expressing a cannabinoid synthase polypeptide, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide.

[00902] The present disclosure provides a modified host cell for expressing a cannabinoid synthase polypeptide, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide; c) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMG-CoA synthase (HMGS) polypeptide; d) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3 -hydroxy-3 -methyl-glutaryl-CoA reductase (tHMGR) polypeptide; e) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate kinase (MK) polypeptide; f) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a phosphomevalonate kinase (PMK) polypeptide; g) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate pyrophosphate decarboxylase (MVD1) polypeptide; h) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a isopentenyl diphosphate isomerase (IDI1) polypeptide; i) one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide; j) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a pyruvate decarboxylase (PDC) polypeptide; and k) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate synthetase (GPPS) polypeptide.

[00903] The present disclosure provides a modified host cell for expressing a cannabinoid synthase polypeptide, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide; c) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMG-CoA synthase (HMGS) polypeptide; d) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3 -hydroxy-3 -methyl-glutaryl-CoA reductase (tHMGR) polypeptide; e) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate kinase (MK) polypeptide; f) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a phosphomevalonate kinase (PMK) polypeptide; g) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate pyrophosphate decarboxylase (MVD1) polypeptide, h) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a isopentenyl diphosphate isomerase (IDI1) polypeptide; i) one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide; and j) one or more

heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate synthetase (GPPS) polypeptide.

[00904] The present disclosure provides a modified host cell for expressing a cannabinoid synthase polypeptide, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide; and c) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides. In certain such embodiments, the modified host cell for expressing a cannabinoid synthase polypeptide comprises a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides.

[00905] The present disclosure provides a modified host cell for expressing a cannabinoid synthase polypeptide, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and b) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides. In certain such embodiments, the modified host cell for expressing a cannabinoid synthase polypeptide comprises a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides.

[00906] The present disclosure provides a modified host cell for expressing a cannabinoid synthase polypeptide, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide; c) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMG-CoA synthase (HMGS) polypeptide; d) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3 -hydroxy-3 -methyl-glutaryl-CoA reductase (tHMGR) polypeptide; e) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate kinase (MK) polypeptide; f) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a phosphomevalonate kinase (PMK) polypeptide; g) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate pyrophosphate decarboxylase (MVD1) polypeptide; h) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a isopentenyl diphosphate isomerase (IDI1) polypeptide; i) one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide; j) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a pyruvate decarboxylase (PDC) polypeptide; k) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate synthetase (GPPS) polypeptide; and 1) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides. In certain such embodiments, the modified host cell for expressing a cannabinoid synthase polypeptide comprises a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides. [00907] The present disclosure provides a modified host cell for expressing a cannabinoid synthase polypeptide, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide; c) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMG-CoA synthase (HMGS) polypeptide; d) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3 -hydroxy-3 -methyl-glutaryl-CoA reductase (tHMGR) polypeptide; e) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate kinase (MK) polypeptide; f) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a phosphomevalonate kinase (PMK) polypeptide; g) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate pyrophosphate decarboxylase (MVD1) polypeptide; h) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a isopentenyl diphosphate isomerase (IDI1) polypeptide; i) one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide; j) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate synthetase (GPPS) polypeptide; and k) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides. In certain such embodiments, the modified host cell for expressing a cannabinoid synthase polypeptide comprises a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides.

[00908] The present disclosure provides a modified host cell for expressing a cannabinoid synthase polypeptide, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide; and c) a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides. In certain such embodiments, the modified host cell for expressing a cannabinoid synthase polypeptide comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides.

[00909] The present disclosure provides a modified host cell for expressing a cannabinoid synthase polypeptide, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and b) a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides. In certain such embodiments, the modified host cell for expressing a cannabinoid synthase polypeptide comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides.

[00910] The present disclosure provides a modified host cell for expressing a cannabinoid synthase polypeptide, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide; c) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMG-CoA synthase (HMGS) polypeptide; d) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3 -hydroxy-3 -methyl-glutaryl-CoA reductase (tHMGR) polypeptide; e) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate kinase (MK) polypeptide; f) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a phosphomevalonate kinase (PMK) polypeptide; g) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate pyrophosphate decarboxylase (MVD1) polypeptide; h) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a isopentenyl diphosphate isomerase (IDI1) polypeptide; i) one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide; j) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a pyruvate decarboxylase (PDC) polypeptide; k) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate synthetase (GPPS) polypeptide; and 1) a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides. In certain such embodiments, the modified host cell for expressing a cannabinoid synthase polypeptide comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides.

[00911] The present disclosure provides a modified host cell for expressing a cannabinoid synthase polypeptide, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide; c) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMG-CoA synthase (HMGS) polypeptide; d) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3 -hydroxy-3 -methyl-glutaryl-CoA reductase (tHMGR) polypeptide; e) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate kinase (MK) polypeptide; f) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a phosphomevalonate kinase (PMK) polypeptide; g) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate pyrophosphate decarboxylase (MVD1) polypeptide; h) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a isopentenyl diphosphate isomerase (IDI1) polypeptide; i) one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide; j) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate synthetase (GPPS) polypeptide; and k) a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides. In certain such embodiments, the modified host cell for expressing a cannabinoid synthase polypeptide comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides.

[00912] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides. In certain such embodiments, the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin mononucleotide (FMN) synthetase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosidase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosyltransferase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more peptidyl-prolyl isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more fatty acid desaturase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein transport and trafficking polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more signal peptidase or signal peptidase complex polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR), one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in endoplasmic reticulum-associated degradation (ERAD), one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cell wall assembly, or one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in vacuolar protein sorting, or a combination of any of the foregoing.

[00913] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a JEM1 polypeptide and a KAR2 polypeptide. In some embodiments, the one or more chaperone or co-chaperone

polypeptides comprise a PFD2s polypeptide. In some embodiments, the one or more chaperone or co-chaperone polypeptides comprise a IEM1 polypeptide. In some

embodiments, the one or more chaperone or co-chaperone polypeptides comprise a CNE1 polypeptide. In some embodiments, the one or more chaperone or co-chaperone polypeptides comprise a SCI1 polypeptide. In some embodiments, the one or more chaperone or co chaperone polypeptides comprise a SIL1 polypeptide. In some embodiments, the one or more chaperone or co-chaperone polypeptides comprise a FPRl polypeptide. In some embodiments, the one or more chaperone or co-chaperone polypeptides comprise a ROT1 polypeptide.

[00914] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum. In certain such embodiments, the one or more polypeptides involved in protein translocation into the endoplasmic reticulum comprise a SEC61 polypeptide, a SBH1 polypeptide, and a SSS1 polypeptide. In some embodiments, the one or more polypeptides involved in protein translocation into the endoplasmic reticulum comprise a SRP14 polypeptide, a SRP21 polypeptide, a SRP68 polypeptide, a SRP72 polypeptide, and a SRP54 polypeptide. In certain such embodiments, the modified host cell for expressing a cannabinoid synthase polypeptide comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding SRP protein complex RNA component SCR1. In some embodiments, the one or more polypeptides involved in protein translocation into the endoplasmic reticulum comprise a SRP101 polypeptide and a SRP102 polypeptide.

[00915] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR). In certain such embodiments, the one or more chaperone or co chaperone polypeptides comprise KAR2 and the one or more polypeptides involved in UPR comprise a HACls polypeptide.

[00916] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR). In certain such embodiments, the one or more chaperone or co- chaperone polypeptides comprise a KAR2 polypeptide and the one or more polypeptides involved in UPR comprise an IRE1 polypeptide.

[00917] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR), one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides. In certain such embodiments, the one or more chaperone or co chaperone polypeptides comprise a KAR2 polypeptide, the one or more polypeptides involved in UPR comprise an IRE1 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, and the one or more thiol oxidase polypeptides comprise an EROl polypeptide.

[00918] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide and a DED1 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, and the one or more thiol oxidase polypeptides comprise an EROl polypeptide.

[00919] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR), one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides. In certain such embodiments, the one or more chaperone or co chaperone polypeptides comprise a KAR2 polypeptide, the one or more polypeptides involved in UPR comprise a PPQ1 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, and the one or more thiol oxidase polypeptides comprise an EROl polypeptide.

[00920] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein transport and trafficking polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide and the one or more protein transport and trafficking polypeptides comprise a BFR2 polypeptide.

[00921] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR) In certain such embodiments, the one or more polypeptides involved in UPR comprise a HACls polypeptide and/or an IRE1 polypeptide. In some embodiments, the one or more polypeptides involved in UPR comprise a HACls polypeptide In some embodiments, the one or more polypeptides involved in unfolded protein response (UPR) comprise an IRE1 polypeptide.

[00922] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, and the one or more thiol oxidase polypeptides comprise an EROl polypeptide.

[00923] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, and the one or more thiol oxidase polypeptides comprise an ERV2 polypeptide.

[00924] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more thiol oxidase polypeptides comprise an EROl polypeptide, and the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide.

[00925] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in UPR, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide and a SSB1 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more thiol oxidase polypeptides comprise an EROl polypeptide, the one or more polypeptides involved in UPR comprise a HACls polypeptide, and the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide. In certain such embodiments, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide, wherein the signal sequence polypeptide is a vacuolar localization signal sequence polypeptide, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide or a PRClt polypeptide.

[00926] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in ERAD, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more polypeptides involved in ERAD comprise a NPL4 polypeptide, and the one or more thiol oxidase polypeptides comprise an EROl polypeptide.

[00927] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cell wall assembly, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more polypeptides involved in cell wall assembly comprise a KRE1 polypeptide, and the one or more thiol oxidase polypeptides comprise an EROl polypeptide.

[00928] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in vacuolar protein sorting, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more polypeptides involved in vacuolar protein sorting comprise a PEP1 polypeptide, and the one or more thiol oxidase polypeptides comprise an EROl polypeptide. [00929] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein transport and trafficking polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more protein transport and trafficking polypeptides comprise a LDB17 polypeptide, and the one or more thiol oxidase polypeptides comprise an EROl polypeptide.

[00930] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides. In certain such embodiments, the one or more chaperone or co chaperone polypeptides comprise a KAR2 polypeptide, a PFD1 polypeptide, and a PFD2s polypeptide; the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide; the one or more polypeptides involved in protein translocation into the endoplasmic reticulum comprise a SRP14 polypeptide, a SRP21 polypeptide, a SRP68 polypeptide, a SRP72 polypeptide, a SEC65 polypeptide, and a SRP54 polypeptide; and the one or more thiol oxidase polypeptides comprise an EROl polypeptide. In certain such embodiments, the modified host cell for expressing a cannabinoid synthase polypeptide comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding SRP protein complex RNA component SCR1.

[00931] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides. In certain such embodiments, the one or more chaperone or co chaperone polypeptides comprise a KAR2 polypeptide; the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide; the one or more polypeptides involved in protein translocation into the endoplasmic reticulum comprise a SRP14 polypeptide, a SRP21 polypeptide, a SRP68 polypeptide, a SRP72 polypeptide, a SEC65 polypeptide, a SRP54 polypeptide, a SEC61 polypeptide, a SBH1 polypeptide, and a SSS1 polypeptide, and the one or more thiol oxidase polypeptides comprise an EROl polypeptide. In certain such embodiments, the modified host cell for expressing a cannabinoid synthase polypeptide comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding SRP protein complex RNA component SCR1.

[00932] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides. In certain such embodiments, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide and the one or more thiol oxidase polypeptides comprise an EROl polypeptide and an ERV2 polypeptide.

[00933] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR). In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide and a SSB1 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more thiol oxidase polypeptides comprise an EROl polypeptide, and the one or more polypeptides involved in UPR comprise a HACls polypeptide.

[00934] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides. In certain such embodiments, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide and the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide.

[00935] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin mononucleotide (FMN) synthetase polypeptides and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides. In certain such embodiments, the one or more flavin

[00936] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosidase polypeptides. In certain such embodiments, the one or more glycosidase polypeptides comprise a CWH41 polypeptide.

[00937] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cell wall assembly. In certain such embodiments, the one or more polypeptides involved in cell wall assembly comprise a KRE1 polypeptide.

[00938] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in ERAD. In certain such embodiments, the one or more polypeptides involved in ERAD comprise a NPL4 polypeptide.

[00939] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, and the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide.

[00940] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide and the one or more protein disulfide isomerase polypeptides comprise a PDIl polypeptide.

[00941] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein transport and trafficking polypeptides. In certain such embodiments, the one or more protein transport and trafficking polypeptides comprise a SEC7 polypeptide. In some embodiments, the one or more protein transport and trafficking polypeptides comprise a BFR2 polypeptide. In some embodiments, the one or more protein transport and trafficking polypeptides comprise a SEC23 polypeptide, a SEC24 polypeptide, a SEC13 polypeptide, a SEC31 polypeptide, a SAR1 polypeptide, a SEC 12 polypeptide, a SEC4 polypeptide, a SEC 16 polypeptide, and an ERV29 polypeptide. In some embodiments, the one or more protein transport and trafficking polypeptides comprise a SEC5 polypeptide, a SEC22 polypeptide, a BOS1 polypeptide, a BETl polypeptide, a BUGl polypeptide, a GRH1 polypeptide, a USOl polypeptide, a SEC 17 polypeptide, a SEC 18 polypeptide, a SSOl polypeptide, a SEC9 polypeptide, and a SNC2 polypeptide.

[00942] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more signal peptidase or signal peptidase complex polypeptides. In certain such embodiments, the one or more signal peptidase or signal peptidase complex polypeptides comprise a KEX2 polypeptide. In some embodiments, the one or more signal peptidase or signal peptidase complex polypeptides comprise a SPC1 polypeptide, a SPC2 polypeptide, a SPC3 polypeptide, and a SEC11 polypeptide.

[00943] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein transport and trafficking polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide and the one or more protein transport and trafficking polypeptides comprise a SEC24 polypeptide.

[00944] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides. In certain such embodiments, the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more glycosidase polypeptides, one or more genes encoding one or more glycosyltransferase polypeptides, one or more genes encoding one or more polypeptides involved in lipid droplet assembly, one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism, or one or more genes encoding one or more vacuolar proteinase polypeptides, or a combination of any of the foregoing. In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises a deletion of one or more genes encoding one or more secretory pathway polypeptides. In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises a downregulation of one or more genes encoding one or more secretory pathway polypeptides.

[00945] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more glycosidase polypeptides and one or more genes encoding one or more polypeptides involved in lipid droplet assembly. In certain such embodiments, the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene and the one or more genes encoding one or more polypeptides involved in lipid droplet assembly comprise a FLD1 gene.

[00946] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more polypeptides involved in lipid droplet assembly. In certain such embodiments, the one or more genes encoding one or more polypeptides involved in lipid droplet assembly comprise a FLD1 gene.

[00947] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more glycosidase polypeptides. In certain such embodiments, the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 and/or MNS1 gene. In some embodiments, the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene. In some embodiments, the one or more genes encoding one or more glycosidase polypeptides comprise a MNS1 gene. [00948] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more glycosyltransferase polypeptides. In certain such embodiments, the one or more genes encoding one or more glycosyltransferase polypeptides comprise an ALG12 gene.

[00949] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides. In certain such embodiments, the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene.

[00950] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides. In certain such embodiments, the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin mononucleotide (FMN) synthetase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosidase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosyltransferase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more peptidyl-prolyl isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more fatty acid desaturase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein transport and trafficking polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more signal peptidase or signal peptidase complex polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR), one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in endoplasmic reticulum-associated degradation (ERAD), one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cell wall assembly, or one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in vacuolar protein sorting, or a combination of any of the foregoing, and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more glycosidase polypeptides, one or more genes encoding one or more glycosyltransferase polypeptides, one or more genes encoding one or more polypeptides involved in lipid droplet assembly, one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism, or one or more genes encoding one or more vacuolar proteinase polypeptides, or a combination of any of the foregoing. In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more

heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and a deletion of one or more genes encoding one or more secretory pathway polypeptides. In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more

heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and a downregulation of one or more genes encoding one or more secretory pathway polypeptides.

[00951] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more polypeptides involved in lipid droplet assembly. In certain such embodiments, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide, and the one or more genes encoding one or more polypeptides involved in lipid droplet assembly comprise a FLDl gene.

[00952] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more glycosidase polypeptides. In certain such embodiments, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide, and the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene.

[00953] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more glycosidase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide, and the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene.

[00954] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more glycosidase polypeptides and one or more genes encoding one or more vacuolar proteinase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene, and the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene.

[00955] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR), and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more glycosidase polypeptides and one or more genes encoding one or more vacuolar proteinase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more polypeptides involved in unfolded protein response (UPR) comprise an IRE1 polypeptide, the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene, and the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene.

[00956] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more glycosidase polypeptides and one or more genes encoding one or more polypeptides involved in lipid droplet assembly. In certain such embodiments, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide, the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene, and the one or more genes encoding one or more polypeptides involved in lipid droplet assembly comprise a FLD1 gene.

[00957] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more glycosidase polypeptides, one or more genes encoding one or more polypeptides involved in lipid droplet assembly, and one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism. In certain such embodiments, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FADl polypeptide, the one or more thiol oxidase polypeptides comprise an EROl polypeptide, the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene, the one or more genes encoding one or more polypeptides involved in lipid droplet assembly comprise a FLD1 gene, and the one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism comprise an OPI1 gene.

[00958] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides and one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a CPR5 polypeptide and a KAR2 polypeptide, the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide, the one or more thiol oxidase polypeptides comprise an EROl polypeptide, the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene, and the one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism comprise an OPI1 gene. In certain such embodiments, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide, wherein the signal sequence polypeptide is a vacuolar localization signal sequence polypeptide, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide.

[00959] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a CPR5 polypeptide and the one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism comprise an OPI1 gene. In certain such embodiments, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide, wherein the signal sequence polypeptide is a vacuolar localization signal sequence polypeptide, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide.

[00960] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides, one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism, and one or more genes encoding one or more glycosidase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a CPR5 polypeptide and a KAR2 polypeptide, the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide, the one or more thiol oxidase polypeptides comprise an EROl polypeptide, the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene, the one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism comprise an OPI1 gene, and the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene. In certain such embodiments, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide, wherein the signal sequence polypeptide is a vacuolar localization signal sequence polypeptide, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide.

[00961] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR), and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides, one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism, and one or more genes encoding one or more glycosidase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a CPR5 polypeptide and a KAR2 polypeptide, the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide, the one or more thiol oxidase polypeptides comprise an EROl polypeptide, the one or more polypeptides involved in unfolded protein response (UPR) comprise an IRE1 polypeptide, the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene, the one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism comprise an OPI1 gene, and the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene. In certain such embodiments, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide, wherein the signal sequence polypeptide is a vacuolar localization signal sequence polypeptide, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide.

[00962] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR), and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides, one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism, and one or more genes encoding one or more glycosidase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a CPR5 polypeptide, a DED1 polypeptide, and a KAR2 polypeptide, the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide, the one or more thiol oxidase polypeptides comprise an EROl polypeptide, the one or more polypeptides involved in unfolded protein response (UPR) comprise an IRE1 polypeptide, the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene, the one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism comprise an OPI1 gene, and the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene. In certain such embodiments, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide, wherein the signal sequence polypeptide is a vacuolar localization signal sequence polypeptide, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide.

[00963] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR), and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides and one or more genes encoding one or more glycosidase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide and a DED1 polypeptide, the one or more thiol oxidase polypeptides comprise an EROl polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more polypeptides involved in unfolded protein response (UPR) comprise an IRE1 polypeptide, the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene, and the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene.

[00964] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR), and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides, one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism, and one or more genes encoding one or more glycosidase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a CPR5 polypeptide and a KAR2 polypeptide, the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide, the one or more thiol oxidase polypeptides comprise an EROl polypeptide, the one or more polypeptides involved in unfolded protein response (UPR) comprise an IRE1 polypeptide and a PPQ1 polypeptide, the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene, the one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism comprise an OPI1 gene, and the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene. In certain such embodiments, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide, wherein the signal sequence polypeptide is a vacuolar localization signal sequence polypeptide, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide.

[00965] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in vacuolar protein sorting, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR), and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides, one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism, and one or more genes encoding one or more glycosidase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a CPR5 polypeptide and a KAR2 polypeptide, the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide, the one or more thiol oxidase polypeptides comprise an ERO 1 polypeptide, the one or more polypeptides involved in vacuolar protein sorting comprise a PEP1 polypeptide, the one or more polypeptides involved in unfolded protein response (UPR) comprise an IRE1 polypeptide, the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene, the one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism comprise an OPI1 gene, and the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene. In certain such embodiments, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide, wherein the signal sequence polypeptide is a vacuolar localization signal sequence polypeptide, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide.

[00966] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in vacuolar protein sorting, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR), and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides and one or more genes encoding one or more glycosidase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide, the one or more thiol oxidase polypeptides comprise an EROl polypeptide, the one or more polypeptides involved in vacuolar protein sorting comprise a PEP1 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more polypeptides involved in unfolded protein response (UPR) comprise an IRE1 polypeptide, the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene, and the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene.

[00967] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein transport and trafficking polypeptides, and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides and one or more genes encoding one or more glycosidase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more protein transport and trafficking polypeptides comprise a SEC24 polypeptide, the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene, and the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene. [00968] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR), and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides and one or more genes encoding one or more glycosidase polypeptides. In certain such embodiments, the one or more chaperone or co- chaperone polypeptides comprise a KAR2 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more thiol oxidase polypeptides comprise an EROl polypeptide, the one or more polypeptides involved in unfolded protein response (UPR) comprise an IRE1 polypeptide, the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene, and the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene.

[00969] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR), and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides and one or more genes encoding one or more glycosidase polypeptides. In certain such embodiments, the one or more chaperone or co- chaperone polypeptides comprise a KAR2 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more thiol oxidase polypeptides comprise an ERV2 polypeptide, the one or more polypeptides involved in unfolded protein response (UPR) comprise an IRE1 polypeptide, the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene, and the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene.

[00970] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, the one or more thiol oxidase polypeptides comprise an EROl polypeptide, and the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene.

[00971] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide, the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide, and the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene.

[00972] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein transport and trafficking polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR), and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides, one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism, and one or more genes encoding one or more glycosidase polypeptides. In certain such embodiments, the one or more chaperone or co chaperone polypeptides comprise a CPR5 polypeptide and a KAR2 polypeptide, the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide, the one or more thiol oxidase polypeptides comprise an EROl polypeptide, the one or more protein transport and trafficking polypeptides comprise a SEC24 polypeptide, the one or more polypeptides involved in unfolded protein response (EIPR) comprise an IREl polypeptide, the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene, the one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism comprise an OPI1 gene, and the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene. In certain such embodiments, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide, wherein the signal sequence polypeptide is a vacuolar localization signal sequence polypeptide, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide.

[00973] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, and one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR), and the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides, one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism, and one or more genes encoding one or more glycosidase polypeptides. In certain such embodiments, the one or more chaperone or co-chaperone polypeptides comprise a CPR5 polypeptide and a KAR2 polypeptide, the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide, the one or more thiol oxidase polypeptides comprise an EROl polypeptide and an ERY2 polypeptide, the one or more polypeptides involved in unfolded protein response (UPR) comprise an IRE1 polypeptide, the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene, the one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism comprise an OPI1 gene, and the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene. In certain such embodiments, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide, wherein the signal sequence polypeptide is a vacuolar localization signal sequence polypeptide, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide.

[00974] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises:

b) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise:

c) deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise:

In certain such embodiments, the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[00975] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises: a) two heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide;

4) a heterologous nucleic acid comprising a nucleotide sequence encoding a protein disulfide isomerase polypeptide, wherein the protein disulfide isomerase polypeptide is a PDIl polypeptide; and

[00976] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises:

[00977] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises:

[00978] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises:

1) a gene encoding a glycosidase polypeptide, wherein the gene encoding the glycosidase polypeptide is a ROT2 gene; and 2) a gene encoding a vacuolar proteinase polypeptide, wherein the gene encoding the vacuolar proteinase polypeptide is a PEP4 gene.

[00979] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises:

[00980] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises:

a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; and

[00981] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises:

a) a heterologous nucleic acid comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; and

[00982] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises:

[00983] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises:

a) a heterologous nucleic acid comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; and b) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise:

[00984] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises:

2) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a chaperone or co-chaperone polypeptide, wherein the chaperone or co-chaperone polypeptide is a KAR2 polypeptide; 3) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a flavin adenine dinucleotide (FAD) synthetase polypeptide, wherein the flavin adenine dinucleotide (FAD) synthetase polypeptide is a FAD1 polypeptide;

[00985] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises:

3) a heterologous nucleic acid comprising a nucleotide sequence encoding a flavin adenine dinucleotide (FAD) synthetase polypeptide, wherein the flavin adenine dinucleotide (FAD) synthetase polypeptide is a FADl polypeptide; 4) a heterologous nucleic acid comprising a nucleotide sequence encoding a protein disulfide isomerase polypeptide, wherein the protein disulfide isomerase polypeptide is a PDIl polypeptide;

[00986] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises:

a) two or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; and

[00987] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises:

a) two heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide; and

2) a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in unfolded protein response, wherein the polypeptide involved in unfolded protein response is a HACl s polypeptide.

[00988] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises two or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide. In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises two heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide.

[00989] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises two or more heterologous nucleic acids comprising a nucleotide sequence encoding a tHMGR polypeptide. In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises two heterologous nucleic acids comprising a nucleotide sequence encoding a tHMGR polypeptide.

[00990] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises two or more heterologous nucleic acids comprising a nucleotide sequence encoding a KAR2 polypeptide. In some

embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises three or more heterologous nucleic acids comprising a nucleotide sequence encoding a KAR2 polypeptide. In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises four or more heterologous nucleic acids comprising a nucleotide sequence encoding a KAR2 polypeptide. In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises two or more heterologous nucleic acids comprising a nucleotide sequence encoding an EROl polypeptide. In some

embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises two or more heterologous nucleic acids comprising a nucleotide sequence encoding a PDIl polypeptide.

[00991] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises two heterologous nucleic acids comprising a nucleotide sequence encoding a KAR2 polypeptide. In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises three heterologous nucleic acids comprising a nucleotide sequence encoding a KAR2 polypeptide. In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises four heterologous nucleic acids comprising a nucleotide sequence encoding a KAR2 polypeptide. In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises two heterologous nucleic acids comprising a nucleotide sequence encoding an EROl polypeptide. In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the modified host cell comprises two heterologous nucleic acids comprising a nucleotide sequence encoding a PDI1 polypeptide.

[00992] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the nucleotide sequence encoding the cannabinoid synthase polypeptide is codon-optimized.

[00993] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide. In certain such embodiments, the one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide comprise a nucleotide sequence encoding a signal sequence polypeptide. In some embodiments, the signal sequence polypeptide is a secretory signal sequence polypeptide. In some

embodiments, the secretory signal sequence polypeptide is a native secretory signal sequence polypeptide. In some embodiments, the secretory signal sequence polypeptide is a synthetic secretory signal sequence polypeptide. In some embodiments, the secretory signal sequence polypeptide is an endoplasmic reticulum retention signal sequence polypeptide. In certain such embodiments, the endoplasmic reticulum retention signal sequence polypeptide is a HDEL polypeptide or a KDEL polypeptide. In some embodiments, the secretory signal sequence polypeptide is a mitochondrial targeting signal sequence polypeptide. In some embodiments, the secretory signal sequence polypeptide is a Golgi targeting signal sequence polypeptide. In some embodiments, the secretory signal sequence polypeptide is a vacuolar localization signal sequence polypeptide. In certain such embodiments, the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide or a PRClt polypeptide. In certain such embodiments, the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide. In some embodiments, the secretory signal sequence polypeptide is a plasma membrane localization signal sequence polypeptide. In some embodiments, the secretory signal sequence polypeptide is a peroxisome targeting signal sequence polypeptide. In some embodiments, the peroxisome targeting signal sequence polypeptide is a PEX8 polypeptide. In some embodiments, the secretory signal sequence polypeptide is a mating factor secretory signal sequence polypeptide (e.g., a MF polypeptide or an evolved MF polypeptide (MFev)). In some embodiments, the signal sequence polypeptide is linked to the N-terminus of the cannabinoid synthase polypeptide.

[00994] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the cannabinoid synthase polypeptide is a fusion polypeptide with an AGA2t polypeptide. In certain such embodiments, the one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide comprise a nucleotide sequence encoding an AGA2t polypeptide. In certain such embodiments, the modified host cell for expressing a cannabinoid synthase polypeptide is modified with one or more heterologous nucleic acids comprising a nucleotide sequence encoding an AGA1 polypeptide. In certain such embodiments, the cannabinoid synthase- AGA2t fusion polypeptide may be displayed on the cell surface of the modified host cell. In some embodiments, the AGA2t polypeptide is linked to the N-terminus of the cannabinoid synthase polypeptide.

[00995] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the cannabinoid synthase polypeptide is a fusion polypeptide with a GFP polypeptide. In certain such embodiments, the one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide comprise a nucleotide sequence encoding a GFP polypeptide. In some embodiments, the GFP polypeptide is linked to the N-terminus of the cannabinoid synthase polypeptide.

Suitable Host Cells

[00996] Parent host cells that are suitable for use in generating a modified host cell of the present disclosure may include eukaryotic cells. In some embodiments, the eukaryotic cells are yeast cells.

[00997] Host cells (including parent host cells and modified host cells) are in some embodiments unicellular organisms, or are grown in culture as single cells. In some embodiments, the host cell is a eukaryotic cell. Suitable eukaryotic host cells may include, but are not limited to, yeast cells and fungal cells. Suitable eukaryotic host cells may include, but are not limited to, Pichia pastoris (now known as Komagataella phaffii ), Pichia finlandica , Pichia trehalophila , Pichia koclamae , Pichia membranaefaciens , Pichia opuntiae, Pichia thermotolerans, Pichia salictaria, Pichia guercuum, Pichia pijperi, Pichia stiptis , Pichia methanolica , Pichia sp., Saccharomyces cerevisiae, Saccharomyces sp., Hansenula polymorpha (now known as Pichia angusta ), Yarrowia lipolytica , Kluyveromyces sp., Kluyveromyces lactis, Kluyveromyces marxianus, Schizosaccharomyces pombe, Scheffersomyces stipites, Dekkera bruxellensis , Blastobotrys adeninivorans (formerly Arxula adeninivorans ), Candida albicans , Aspergillus nidulans , Aspergillus niger , Aspergillus oryzae, Trichoderma reesei, Chrysosporium lucknowense, Fusarium sp., Fusarium gramineum , Fusarium venenatum , Neurospora crassa , and the like. In some embodiments, the modified host cell disclosed herein is cultured in vitro.

[00998] In some embodiments, the host cell of the disclosure is a yeast cell. In some embodiments, the host cell is a protease-deficient strain of Saccharomyces cerevisiae. In some embodiments, the host cell is Saccharomyces cerevisiae. In some embodiments, the host cell for use in generating a modified host cell of the present disclosure may be selected because of ease of culture; rapid growth; availability of tools for modification, such as promoters and vectors; and the host cell’s safety profile. In some embodiments, the host cell for use in generating a modified host cell of the present disclosure may be selected because of its ability or inability to introduce certain posttranslational modifications onto expressed polypeptides, such as cannabinoid synthase polypeptides. For instance, modified

Komagataella phaffii host cells may hyperglycosylate cannabinoid synthase polypeptides and hyperglycosylation may alter the activity of the resultant expressed polypeptide.

Genetic Modification of Host Cells

[00999] The present disclosure provides for a method of making a modified host cell for producing a cannabinoid or a cannabinoid derivative, the method comprising introducing into a host cell one or more heterologous nucleic acids disclosed herein. The disclosure provides a method of making a modified host cell for producing a cannabinoid or a cannabinoid derivative, the method comprising introducing into a host cell: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and b) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides. In some embodiments, the nucleotide sequence encoding the cannabinoid synthase polypeptide is codon-optimized. In some embodiments, the nucleotide sequences encoding the secretory pathway polypeptides are codon-optimized. In some embodiments, the modified host cell for producing a cannabinoid or a cannabinoid derivative comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides.

[001000] In some embodiments, the modified host cell for producing a cannabinoid or a cannabinoid derivative comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides. The disclosure provides a method of making a modified host cell for producing a cannabinoid or a cannabinoid derivative, the method comprising introducing into a host cell: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and b) a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides.

[001001] In some embodiments, the modified host cell for producing a cannabinoid or a cannabinoid derivative comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides. The disclosure provides a method of making a modified host cell for producing a cannabinoid or a cannabinoid derivative, the method comprising introducing into a host cell: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide, b) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, and c) a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides.

[001002] In some embodiments, the modified host cell for producing a cannabinoid or a cannabinoid derivative may comprise one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and express or overexpress combinations of heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cannabinoid or cannabinoid precursor (e.g., geranylpyrophosphate (GPP), prenyl phosphates, olivetolic acid, or hexanoyl-CoA) biosynthesis. In some embodiments, the methods of making a modified host cell for producing a cannabinoid or a cannabinoid derivative comprise introducing into a host cell one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cannabinoid or cannabinoid precursor biosynthesis. In some embodiments disclosed herein, the nucleotide sequences encoding the one or more polypeptides involved in cannabinoid or cannabinoid precursor biosynthesis are codon- optimized.

[001003] In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide, such as a secretory signal sequence polypeptide. In some

embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the cannabinoid synthase polypeptide is a fusion polypeptide with an AGA2t polypeptide. In some embodiments of the modified host cell for producing a cannabinoid or a cannabinoid derivative, the cannabinoid synthase polypeptide is a fusion polypeptide with a GFP polypeptide.

[001004] The present disclosure provides for a method of making a modified host cell for expressing a cannabinoid synthase polypeptide, the method comprising introducing into a host cell one or more heterologous nucleic acids disclosed herein. The disclosure provides a method of making a modified host cell for expressing a cannabinoid synthase polypeptide, the method comprising introducing into a host cell: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and b) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides. In some embodiments, the nucleotide sequence encoding the cannabinoid synthase polypeptide is codon-optimized. In some embodiments, the nucleotide sequences encoding the secretory pathway polypeptides are codon-optimized. In some embodiments, the modified host cell for expressing a cannabinoid synthase polypeptide comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides.

[001005] In some embodiments, the modified host cell for expressing a cannabinoid synthase polypeptide comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides. The disclosure provides a method of making a modified host cell for expressing a cannabinoid synthase polypeptide, the method comprising introducing into a host cell: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and b) a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides.

[001006] In some embodiments, the modified host cell for expressing a cannabinoid synthase polypeptide comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptide, and a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides. The disclosure provides a method of making a modified host cell for expressing a cannabinoid synthase polypeptide, the method comprising introducing into a host cell: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide, b) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, and c) a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides.

[001007] In some embodiments, the modified host cell expressing a cannabinoid synthase polypeptide may comprise one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and express or overexpress combinations of heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cannabinoid or cannabinoid precursor (e g., geranylpyrophosphate (GPP), prenyl phosphates, olivetolic acid, or hexanoyl-CoA) biosynthesis. In some embodiments, the methods of making a modified host cell expressing a cannabinoid synthase polypeptide comprise introducing into a host cell one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cannabinoid or cannabinoid precursor biosynthesis. In some embodiments disclosed herein, the nucleotide sequences encoding the one or more polypeptides involved in cannabinoid or cannabinoid precursor biosynthesis are codon- optimized.

[001008] In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the cannabinoid synthase polypeptide comprises a signal sequence polypeptide, such as a secretory signal sequence polypeptide. In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the cannabinoid synthase polypeptide is a fusion polypeptide with an AGA2t polypeptide. In some embodiments of the modified host cell for expressing a cannabinoid synthase polypeptide, the cannabinoid synthase polypeptide is a fusion polypeptide with a GFP polypeptide.

[001009] To modify a parent host cell to produce a modified host cell of the present disclosure, one or more heterologous nucleic acids disclosed herein may be introduced stably or transiently into a host cell, using established techniques. Such techniques may include, but are not limited to, electroporation, calcium phosphate precipitation, DEAE-dextran mediated transfection, liposome-mediated transfection, the lithium acetate method, and the like. See Gietz, R.D. and R.A. Woods. (2002) TRANSFORMATION OF YEAST BY THE Liac/SS CARRIER DN A/PEG METHOD. For stable transformation, a plasmid, vector, expression construct, etc. comprising one or more nucleic acids (e.g., heterologous) disclosed herein will generally further include a selectable marker, e g., any of several well-known selectable markers such as neomycin resistance, ampicillin resistance, tetracycline resistance, chloramphenicol resistance, kanamycin resistance, and the like. In some embodiments, the selectable marker gene to provide a phenotypic trait for selection of transformed host cells is dihydrofolate reductase. In some embodiments, a parent host cell is modified to produce a modified host cell of the present disclosure using a CRISPR/Cas9 system to modify a parent host cell with one or more heterologous nucleic acids disclosed herein.

[001010] In some embodiments, varying polypeptide expression level, such as cannabinoid synthase polypeptide expression level, and/or the production of cannabinoids or cannabinoid derivatives in a modified host cell may be done by changing the gene copy number, promoter strength, and/or promoter regulation and/or by use of codon optimization.

[001011] One or more heterologous nucleic acids disclosed herein can be present in an expression vector or construct. Suitable expression vectors may include, but are not limited to, plasmids, yeast plasmids, yeast artificial chromosomes, and any other vectors specific for specific hosts of interest (such as yeast). Thus, for example, one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate pathway gene product(s) is included in any one of a variety of expression vectors for expressing the mevalonate pathway gene product(s). Such vectors may include chromosomal, non- chromosomal, and synthetic DNA sequences.

[001012] The present disclosure provides for a method of making a modified host cell for producing a cannabinoid or a cannabinoid derivative, the method comprising introducing into a host cell one or more vectors disclosed herein. In certain such embodiments, the one or more vectors comprise one or more vectors comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide. In certain such embodiments, the one or more vectors comprise one or more vectors comprising one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides. In some embodiments, the method comprises introducing into the host cell a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides. In some embodiments, the one or more vectors comprise one or more vectors comprising one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cannabinoid or cannabinoid precursor biosynthesis. In some embodiments, the nucleotide sequences are codon-optimized.

[001013] The present disclosure provides for a method of making a modified host cell for expressing a cannabinoid synthase polypeptide, the method comprising introducing into a host cell one or more vectors disclosed herein. In certain such embodiments, the one or more vectors comprise one or more vectors comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide. In certain such embodiments, the one or more vectors comprise one or more vectors comprising one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides. In some embodiments, the method comprises introducing into the host cell a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides. In some embodiments, the one or more vectors comprise one or more vectors comprising one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cannabinoid or cannabinoid precursor biosynthesis. In some embodiments, the nucleotide sequences are codon-optimized

[001014] Numerous additional suitable expression vectors are known to those of skill in the art, and many are commercially available. The following vectors are provided by way of example; for yeast, the low copy CEN ARS and high copy 2 micron plasmids. However, any other plasmid or other vector may be used so long as it is compatible with the host cell.

[001015] In some embodiments, one or more of the heterologous nucleic acids disclosed herein are present in a single expression vector. In some embodiments, two or more of the heterologous nucleic acids disclosed herein are present in a single expression vector. In some embodiments, three or more of the heterologous nucleic acids disclosed herein are present in a single expression vector. In some embodiments, four or more of the heterologous nucleic acids disclosed herein are present in a single expression vector. In some embodiments, five or more of the heterologous nucleic acids disclosed herein are present in a single expression vector. In some embodiments, six or more of the heterologous nucleic acids disclosed herein are present in a single expression vector. In some

embodiments, seven or more of the heterologous nucleic acids disclosed herein are present in a single expression vector.

[001016] In some embodiments, two or more heterologous nucleic acids disclosed herein are in separate expression vectors. In some embodiments, three or more heterologous nucleic acids disclosed herein are in separate expression vectors. In some embodiments, four or more heterologous nucleic acids disclosed herein are in separate expression vectors. In some embodiments, five or more heterologous nucleic acids disclosed herein are in separate expression vectors. In some embodiments, six or more heterologous nucleic acids disclosed herein are in separate expression vectors. In some embodiments, seven or more heterologous nucleic acids disclosed herein are in separate expression vectors. In some embodiments, eight or more heterologous nucleic acids disclosed herein are in separate expression vectors. In some embodiments, nine or more heterologous nucleic acids disclosed herein are in separate expression vectors. In some embodiments, ten or more heterologous nucleic acids disclosed herein are in separate expression vectors.

[001017] In some embodiments, one or more of the heterologous nucleic acids disclosed herein are present in a single expression construct. In some embodiments, two or more of the heterologous nucleic acids disclosed herein are present in a single expression construct. In some embodiments, three or more of the heterologous nucleic acids disclosed herein are present in a single expression construct. In some embodiments, four or more of the heterologous nucleic acids disclosed herein are present in a single expression construct. In some embodiments, five or more of the heterologous nucleic acids disclosed herein are present in a single expression construct. In some embodiments, six or more of the heterologous nucleic acids disclosed herein are present in a single expression construct. In some embodiments, seven or more of the heterologous nucleic acids disclosed herein are present in a single expression construct.

[001018] In some embodiments, two or more heterologous nucleic acids disclosed herein are in separate expression constructs. In some embodiments, three or more heterologous nucleic acids disclosed herein are in separate expression constructs. In some embodiments, four or more heterologous nucleic acids disclosed herein are in separate expression constructs. In some embodiments, five or more heterologous nucleic acids disclosed herein are in separate expression constructs. In some embodiments, six or more heterologous nucleic acids disclosed herein are in separate expression constructs. In some embodiments, seven or more heterologous nucleic acids disclosed herein are in separate expression constructs. In some embodiments, eight or more heterologous nucleic acids disclosed herein are in separate expression constructs. In some embodiments, nine or more heterologous nucleic acids disclosed herein are in separate expression constructs. In some embodiments, ten or more heterologous nucleic acids disclosed herein are in separate expression constructs.

[001019] In some embodiments, one or more of the heterologous nucleic acids disclosed herein is present in a high copy number plasmid, e.g., a plasmid that exists in about 10-50 copies per cell, or more than 50 copies per cell. In some embodiments, one or more of the heterologous nucleic acids disclosed herein is present in a low copy number plasmid. In some embodiments, one or more of the heterologous nucleic acids disclosed herein is present in a medium copy number plasmid. The copy number of the plasmid may be selected to reduce expression of one or more polypeptides disclosed herein, such as a cannabinoid synthase polypeptide. Reducing expression by limiting the copy number of the plasmid may prevent saturation of the secretory pathway leading to possible protein degradation and/or modified host cell death or a loss of modified host cell viability.

[001020] In some embodiments, the modified host cell has one copy of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide disclosed herein. In some embodiments, the modified host cell has two copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide disclosed herein. In some embodiments, the modified host cell has three copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide disclosed herein. In some embodiments, the modified host cell has four copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide disclosed herein. In some embodiments, the modified host cell has five copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide disclosed herein. In some embodiments, the modified host cell has six copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide disclosed herein. In some embodiments, the modified host cell has seven copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide disclosed herein. In some embodiments, the modified host cell has eight copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide disclosed herein. In some embodiments, the modified host cell has nine copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide disclosed herein. In some embodiments, the modified host cell has ten copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide disclosed herein. In some embodiments, the modified host cell has eleven copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide disclosed herein. In some embodiments, the modified host cell has twelve copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide disclosed herein. In some embodiments, the modified host cell has twelve or more copies of a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide disclosed herein.

[001021] Depending on the host/vector or host/construct system utilized, any of a number of suitable transcription and translation control elements, including constitutive and inducible promoters, transcription enhancer elements, transcription terminators, etc. may be used in the expression vector or construct (see e.g., Bitter et al. (1987 ) Methods in

Enzymology, 153 :516-544).

[001022] In some embodiments, the heterologous nucleic acids disclosed herein are operably linked to a promoter. In some embodiments, the promoter is a constitutive promoter. In some embodiments, the promoter is an inducible promoter. In some embodiments, the promoter is functional in a eukaryotic cell. In some embodiments, the promoter can be a strong driver of expression. In some embodiments, the promoter can be a weak driver of expression. In some embodiments, the promoter can be a medium driver of expression. The promoter may be selected to reduce expression of one or more polypeptides disclosed herein, such as a cannabinoid synthase polypeptide. Reducing expression through promoter selection may prevent saturation of the secretory pathway leading to possible protein degradation and/or modified host cell death or a loss of modified host cell viability. Examples of strong constitutive promoters include, but are not limited to: pTDH3 and pFBAl . Examples of medium constitutive promoters include, but are not limited to: pACTl and pCYCl. An example of a weak constitutive promoter includes, but is not limited to: pSLNl . Examples of strong inducible promoters include, but are not limited to: pGALl and pGALlO. An example of a medium inducible promoter includes, but is not limited to:

pGAL7. An example of a weak inducible promoter includes, but is not limited to: pGAL3.

[001023] Non-limiting examples of suitable eukaryotic promoters may include CMV immediate early, HSV thymidine kinase, early and late SV40, LTRs from retrovirus, and mouse metallothionein-I. Selection of the appropriate vector, construct, and promoter is well within the level of ordinary skill in the art. The expression vector or construct may also contain a ribosome binding site for translation initiation and a transcription terminator. The expression vector or construct may also include appropriate sequences for amplifying expression.

[001024] Inducible promoters are well known in the art. Suitable inducible promoters may include, but are not limited to, a tetracycline-inducible promoter; an estradiol inducible promoter, a sugar inducible promoter, e.g, pGall or pSUC2, an amino acid inducible promoter, e.g., pMet25; a metal inducible promoter, e.g., pCupl, a methanol-inducible promoter, e.g., pAOXl, and the like.

[001025] In yeast, a number of vectors or constructs containing constitutive or inducible promoters may be used. For a review see, Current Protocols in Molecular Biology, Vol. 2, 1988, Ed. Ausubel, et al., Greene Publish. Assoc. & Wiley Interscience, Ch. 13; Grant, et al., 1987, Expression and Secretion Vectors for Yeast, in Methods in Enzymology, Eds. Wu & Grossman, 31987, Acad. Press, N.Y., Vol. 153, pp.516-544; Glover, 1986, DNA Cloning, Vol. II, IRL Press, Wash., D.C., Ch. 3; and Bitter, 1987, Heterologous Gene Expression in Yeast, Methods in Enzymology, Eds. Berger & Kimmel, Acad. Press, N.Y., Vol. 152, pp. 673-684; and The Molecular Biology of the Yeast Saccharomyces , 1982, Eds. Strathem et al., Cold Spring Harbor Press, Vols. I and II. A constitutive yeast promoter such as pADH, pTDH3, pFBAl, pACTl, pCYCl, and pSLNl or an inducible promoter such as pGALl, pGALlO, pGAL7, and pGAL3 may be used (Cloning in Yeast, Ch. 3, R. Rothstein In: DNA Cloning Vol. 11, A Practical Approach, Ed. DM Glover, 1986, IRL Press, Wash.,

D C ). Alternatively, vectors may be used which promote integration of foreign DNA sequences into the yeast chromosome. Generally, recombinant expression vectors will include origins of replication and selectable markers permitting transformation of the host cell, e.g., the S. cerevisiae TRP1 gene or a gene cassette encoding resistance to an antibiotic, etc.; and a promoter derived from a highly-expressed gene to direct transcription of the coding sequence. Such promoters can be derived from genetic sequences encoding glycolytic enzymes such as 3-phosphoglycerate kinase (PGK), a-factor, acid phosphatase, or heat shock proteins, among others.

[001026] In some embodiments, one or more nucleic acids (e.g., heterologous) disclosed herein is integrated into the genome of the modified host cell disclosed herein. In some embodiments, one or more nucleic acids (e.g., heterologous) disclosed herein is integrated into a chromosome of the modified host cell disclosed herein. In some embodiments, one or more nucleic acids (e.g., heterologous) disclosed herein remains episomal (i.e., is not integrated into the genome or a chromosome of the modified host cell). In some embodiments, at least one of the one or more heterologous nucleic acids disclosed herein is maintained extrachromosomally (e.g., on a plasmid or artificial chromosome). The gene copy number of one or more genes encoding one or more polypeptides disclosed herein, such as a cannabinoid synthase polypeptide, may be selected to reduce expression of the one or more polypeptides disclosed herein, such as a cannabinoid synthase polypeptide. Reducing expression by limiting the gene copy number may prevent saturation of the secretory pathway leading to possible protein degradation and/or modified host cell death or a loss of modified host cell viability.

[001027] As will be appreciated by the skilled artisan, slight changes in nucleotide sequence do not necessarily alter the amino acid sequence of the encoded polypeptide. It will be appreciated by persons skilled in the art that changes in the identities of nucleotides in a specific gene sequence that change the amino acid sequence of the encoded polypeptide may result in reduced or enhanced effectiveness of the genes and that, in some applications (e.g., anti-sense, co-suppression, or RNAi), partial sequences often work as effectively as full length versions. The ways in which the nucleotide sequence can be varied or shortened are well known to persons skilled in the art, as are ways of testing the effectiveness of the altered genes. In certain embodiments, effectiveness may easily be tested by, for example, conventional gas chromatography. All such variations of the genes are therefore included as part of the present disclosure.

[001028] Genomic deletion of the open reading frame encoding the protein may abolish all expression of a gene. Downregulation of a gene can be accomplished in several ways at the DNA, RNA, or protein level, with the result being a reduction in the amount of active protein in the cell. Truncations of the open reading frame or the introduction of mutations that destabilize the protein or reduce catalytic activity achieve a similar goal, as does fusing a“degron” polypeptide that destabilizes the protein. Engineering of the regulatory regions of the gene can also be used to change gene expression. Alteration of the promoter sequence or replacement with a different promoter is one method. Truncation of the terminator, known as decreased abundance of mRNA perturbation (DAmP), is also known to reduce gene expression. Other methods that reduce the stability of the mRNA include the use of cis- or trans-acting ribozymes, e.g., self-cleaving ribozymes, or RNA elements that recruit an exonuclease, or antisense DNA. RNAi may be used to silence genes in budding yeast strains via import of the required protein factors from other species, e.g., Drosha or Dice

(Drinnenberg et al 2009). Gene expression may also be silenced in S. cerevisiae via recruitment of native or heterologous silencing factors or repressors, which may be accomplished at arbitrary loci using the D-Cas9 CRISPR system (Qi et al 2013). Protein level can also be reduced by engineering the amino acid sequence of the target protein. A variety of degron sequences may be used to target the protein for rapid degradation, including, but not limited to, ubiquitin fusions and N-end rule residues at the amino terminus. These methods may be implemented in a constitutive or conditional fashion.

[001029] In some embodiments, the modified host cell comprises two or more nucleotide sequences linked via a T2A or an Internal Ribosome Entry Site (IRES) element, allowing for co-expression of the sequences under the control of a single promoter and terminator sequence. In some embodiments, the T2A element is introduced between a nucleotide sequence encoding a cannabinoid synthase polypeptide upstream and a nucleotide sequence encoding a reporter polypeptide, such as a GFP polypeptide, downstream, resulting in two polypeptides separated at the GP (glycine-proline) junction at the end of the T2A element. Use of a T2A element in combination with a reporter polypeptide, such as a GFP polypeptide, may allow for screening or selection of modified host cells that are expressing a polypeptide of interest, as any modified host cells positive for the reporter polypeptide should also express the polypeptide of interest because they are both derived from the same transcript. Such screening may allow for rapid assessment of the solubilization and expression of cannabinoid synthase polypeptides in modified host cells comprising one or more modifications to modulate the expression of one or more secretory pathway polypeptides and/or one or more heterologous nucleic acids comprising a codon-optimized nucleotide sequence encoding a cannabinoid synthase polypeptide. In certain such embodiments, the cannabinoid synthase polypeptide may comprise a signal sequence polypeptide.

[001030] The disclosure provides an assay for screening for expression of a cannabinoid synthase polypeptide in a modified host cell disclosed herein, wherein the modified host cell comprises a nucleotide sequence encoding a cannabinoid synthase polypeptide and a nucleotide sequence encoding a reporter polypeptide, wherein a T2A or an IRES element is introduced between the nucleotide sequence encoding the cannabinoid synthase polypeptide upstream and the nucleotide sequence encoding a reporter polypeptide downstream, said assay comprising: a) culturing the modified host cell in a culture medium and b) detecting the reporter polypeptide. In certain such embodiments, the reporter polypeptide is a GFP polypeptide. In certain such embodiments, the GFP polypeptide is detected by fluorescence detection. In certain such embodiments, a T2A element is used. Induction Systems

[001031] To adapt to a constantly changing environment, microbes such as yeast have evolved a wide range of natural inducible promoter systems. Any promoter that is regulated by a small molecule or change in environment (temperature, pH, oxygen level, osmolarity, oxidative damage) can in principle be converted into an inducible system for the expression of heterologous genes. The best known system in S. cerevisiae is the galactose regulon, which is strongly repressed by glucose and activated by galactose. Heterologous genetic pathways under the control of galactose-inducible promoters are regulated in the same way, and thus an engineered strain can be grown in glucose media to build biomass, and then switched to galactose to induce pathway expression. A range of expression levels can be achieved, from very strong pGALl to relatively weak pGAL3. However, galactose may be expensive and a poor carbon source for S. cerevisiae. Therefore, for industrial applications, it may be advantageous to re-engineer the regulon such that the cells can be induced in a non galactose media. The galactose regulon can be modified for this purpose in many ways, including:

• Overexpressing the negative regulator of GAL80, GAL3, from an inducible promoter, e.g., pSUC2-GAL3, such that switching from glucose to sucrose relieves GAL80 expression and activates the pathway.

• Deleting the repressor GAL80 and replacing the native GAL4 cassette with a version under the control of a sucrose inducible promoter, e.g., pSUC2- GAL4, such that expression is induced by a switch from glucose to sucrose.

• Replacing the native GAL80 gene with an inducible version, e.g., pSUC2- GAL80, such that expression is induced by a switch from sucrose to glucose.

[001032] These strategies often require fine-tuning of the activator and repressor levels to achieve the proper dynamics (very low or no expression in the off state, and desired expression level in the on state). There are a variety of ways to fine tune protein expression, including use of protein stabilization or degradation tags (e.g., degrons) or use of temperature sensitive mutants of the activators or regulators. In the examples above, the pSUC2 promoter is used to induce the galactose regulon in sucrose media. However, any inducible promoter can be used for this purpose, or for control of individual genes outside of the context of the galactose regulon. The list below provides some examples:

• Phosphate regulated promoters, e.g., pPH05

• Carbon source regulated promoters, e.g., pADH2

• Amino acid regulated promoters, e.g., pMET25 • Metal ion induced promoters, e g., pCUPl

• Temperature regulated promoters, e.g., pHSPl2, pHSP26

• pH regulated promoters, e g., pHSPl2, pHSP26

• Oxygen level regulated promoters, e g., pDANl

• Oxidative stress regulated promoters, e.g., promoters from AHP1, TRR1, TRX2, TSA1, GPX2, GSH1, GSH2, GLR1, SOD1, or SOD2 genes.

• ER stress regulated promoters, e.g., unfolded protein response element

promoters.

[001033] In addition to these natural examples, there are a variety of synthetic inducible promoter systems. These are generally based on re-arrangement of native or foreign transcriptional elements into a basal promoter scaffold and/or fusions of activator domains and DNA binding domains to create novel transcription factors. Two examples are provided below:

• Estradiol-inducible systems involving fusion of the estradiol receptor to

DNA-binding and transcriptional activation domain, paired with synthetic or native promoters with binding sites.

• tet Trans Activator (tTA) or reverse tet Trans Activator (rtTA) systems paired with tetO-containing promoters.

[001034] In some embodiments, one of the above inducible promoter systems is used in a modified host cell of the disclosure. In some embodiments, the inducible promoter system is a natural inducible promoter system. In some embodiments, the inducible promoter system is a synthetic inducible promoter system. In some embodiments, a suitable media for culturing modified host cells of the disclosure comprises one or more of the inducers disclosed herein. Possible inducers include:

• Phosphate regulated promoters, e.g., pPH05

o KH2PO4

• Carbon source regulated promoters, e.g., pADH2

o Galactose (e.g., pGALl)

o Glucose (e.g., pADH2)

o Sucrose (e.g., pSUC2, pGPHl, pMALl2)

o Maltose (e.g., pMALl2, pMAL32)

• Amino acid regulated promoters, e.g., pMET25

o Methionine (e.g., pMET25)

o Lysine (e.g., pLYS9) o Other amino acids

• Metal ion induced promoters, e.g., pCUPl

o CuS0₄

• Temperature regulated promoters, e.g., pHSPl2, pHSP26

o Change in temperature, e.g., 30 °C to 37 °C

• pH regulated promoters, e g , pHSPl2, pHSP26

o Change in pH, e.g., pH 6 to pH 4

• Oxygen level regulated promoters, e.g., pDANl

o Change in oxygen level, e.g., 20% to 1% dissolved oxygen levels

• Oxidative stress regulated promoters, e.g., pSODl

o Addition of hydrogen peroxide or superoxide-generating drug menadione

• ER stress regulated promoters, e.g., unfolded protein response element promoters. o Tunicamycin, or expression of proteins prone to misfolding (e.g., cannabinoid synthases)

• Estradiol-inducible systems involving fusion of the estradiol receptor to DNA- binding and transcriptional activation domain, paired with synthetic or native promoters with binding sites.

o Estradiol

o Doxycyclin

Codon Usage

[001035] As is well known to those of skill in the art, it is possible to improve the expression of a heterologous nucleic acid in a host organism by replacing the nucleotide sequences coding for a particular amino acid (i.e., a codon) with another codon which is better expressed in the host organism (i.e., codon-optimization). One reason that this effect arises is due to the fact that different organisms show preferences for different codons. In some embodiments, a nucleic acid disclosed herein is modified or optimized such that the nucleotide sequence reflects the codon preference for the particular host cell. For example, the nucleotide sequence will in some embodiments be modified or optimized for yeast codon preference. In some embodiments, a nucleotide sequence disclosed herein is codon- optimized. See, e.g., Bennetzen and Hall (1982) J Biol. Chem. 257(6): 3026-3031.

[001036] Statistical methods have been generated to analyze codon usage bias in various organisms and many computer algorithms have been developed to implement these statistical analyses in the design of codon optimized gene sequences (Lithwick G, Margalit H (2003) Hierarchy of sequence-dependent features associated with prokaryotic translation. Genome Research 13 : 2665-73). Other modifications in codon usage to increase protein expression that are not dependent on codon bias have also been described (Welch et al. (2009). In some embodiments, codon optimization of the nucleotide sequence may result in an increase in the desired polypeptide or enzyme catalytic activity in the modified host cell.

[001037] In some embodiments, the codon usage of a nucleotide sequence is modified or optimized such that the level of translation of the encoded mRNA is decreased. In some embodiments, a codon-optimized nucleotide sequence may be optimized such that the level of translation of the encoded mRNA is decreased. Reducing the level of translation of an mRNA by modifying codon usage may be achieved by modifying the nucleotide sequence to include codons that are rare or not commonly used by the host cell. Codon usage tables for many organisms are available that summarize the percentage of time a specific organism uses a specific codon to encode for an amino acid. Certain codons are used more often than other,“rare” codons. The use of“rare” codons in a nucleotide sequence generally decreases its rate of translation. Thus, e.g., the nucleotide sequence is modified by introducing one or more rare codons, which affect the rate of translation, but not the amino acid sequence of the polypeptide translated. For example, there are six codons that encode for arginine: CGT, CGC, CGA, CGG, AGA, and AGG. In E. coli the codons CGT and CGC are used far more often (encoding approximately 40% of the arginines in E. coli each) than the codon AGG (encoding approximately 2% of the arginines in E. coli). Modifying a CGT codon within the sequence of a gene to an AGG codon would not change the sequence of the polypeptide, but would likely decrease the gene’s rate of translation. In some embodiments, a codon- optimized nucleotide sequence may be optimized such that the rate of translation of the encoded mRNA is decreased. Slowing translation of the mRNA encoded by nucleotide sequences encoding a cannabinoid synthase polypeptide through use of codon-optimized nucleotide sequences encoding a cannabinoid synthase polypeptide may prevent saturation of the secretory pathway leading to possible protein degradation and/or modified host cell death or a loss of modified host cell viability. Without wishing to be bound by theory, slowing translation of the mRNA encoded by nucleotide sequences encoding a cannabinoid synthase polypeptide may improve translocation of the nascent cannabinoid synthase polypeptide chain to the endoplasmic reticulum (ER), assisting in delivery of the nascent cannabinoid synthase polypeptide chain to the secretory pathway. [001038] In some embodiments, a codon-optimized nucleotide sequence may be optimized for expression in a yeast cell. In certain such embodiments, the yeast cell is Saccharomyces cerevisiae.

[001039] Further, it will be appreciated that this disclosure embraces the degeneracy of codon usage as would be understood by one of ordinary skill in the art and illustrated in the following table.

Codon Deeeneracies

Methods of Producing a Cannabinoid or a Cannabinoid Derivative or of Expressing a Cannabinoid Synthase Polypeptide

[001040] The disclosure provides methods for expressing a cannabinoid synthase polypeptide. In certain such embodiments, the methods may comprise culturing a modified host cell of the disclosure in a culture medium. [001041] The present disclosure also provides methods of producing a cannabinoid or a cannabinoid derivative. The methods may involve culturing a modified host cell of the present disclosure in a culture medium. In certain such embodiments, the methods comprise recovering the produced cannabinoid or cannabinoid derivative. The methods may also involve cell-free production of cannabinoids or cannabinoid derivatives using one or more polypeptides disclosed herein expressed or overexpressed by a modified host cell of the disclosure.

[001042] Cannabinoids or cannabinoid derivatives that can be produced with the methods or modified host cells of the present disclosure may include, but are not limited to, cannabichromene (CBC) type (e g., cannabichromenic acid), cannabigerol (CBG) type (e.g., cannabigerolic acid), cannabidiol (CBD) type (e.g., cannabidiolic acid), A⁹-trans- tetrahydrocannabinol (D⁹ -THC) type (e.g., A⁹-tetrahydrocannabinolic acid), A⁸-trans- tetrahydrocannabinol (D⁸ -THC) type, cannabicyclol (CBL) type, cannabielsoin (CBE) type, cannabinol (CBN) type, cannabinodiol (CBND) type, cannabitriol (CBT) type, derivatives of any of the foregoing, and others as listed in Elsohly M.A. and Slade D., Life Sci. 2005 Dec 22;78(5):539-48. Epub 2005 Sep 30. In some embodiments, the cannabinoid or cannabinoid derivative is produced in an amount of more than 100 mg/L culture medium. In some embodiments, the cannabinoid or cannabinoid derivative is produced in an amount of more than 50 mg/L culture medium.

[001043] Cannabinoids or cannabinoid derivatives that can be produced with the methods or modified host cells of the present disclosure may also include, but are not limited to, cannabigerolic acid (CBGA), cannabigerolic acid monomethylether (CBGAM), cannabigerol (CBG), cannabigerol monomethylether (CBGM), cannabigerovarinic acid (CBGVA), cannabigerovarin (CBGV), cannabichromenic acid (CBCA), cannabichromene (CBC), cannabichromevarinic acid (CBCVA), cannabichromevarin (CBCV), cannabidiolic acid (CBDA), cannabidiol (CBD), cannabidiol monomethylether (CBDM), cannabidiol-C4 (CBD-C4), cannabidivarinic acid (CBDVA), cannabidivarin (CBDV), cannabidiorcol (CBD- Ci), D⁹ -tetrahydrocannabinolic acid A (THCA-A), D⁹ -tetrahydrocannabinolic acid B (THCA-B), D⁹ -tetrahydrocannabinol (THC), D⁹ -tetrahydrocannabinolic acid-C4 (THCA- C4), D⁹ -tetrahydrocannabinol-C4 (THC-C4), D⁹ -tetrahydrocannabivarinic acid (THCYA),

D⁹ -tetrahydrocannabivarin (THCV), D⁹ -tetrahydrocannabiorcolic acid (THCA-Ci), D⁹ - tetrahydrocannabiorcol (THC-Ci), D⁷ -cis-iso-tetrahydrocannabivarin, D⁸ - tetrahydrocannabinolic acid (D⁸ -THCA), D⁸ -tetrahydrocannabinol (D⁸ -THC), cannabicyclolic acid (CBLA), cannabicyclol (CBL), cannabicyclovarin (CBLV), cannabielsoic acid A (CBEA-A), cannabielsoic acid B (CBEA-B), cannabielsoin (CBE), cannabielsoinic acid, cannabicitranic acid, cannabinolic acid (CBNA), cannabinol (CBN), cannabinol methylether (CBNM), cannabinol -C4, (CBN-C4), cannabivarin (CBV), cannabinol-C2 (CNB-C2), cannabiorcol (CBN-Ci), cannabinodiol (CBND), cannabinodivarin (CBVD), cannabitriol (CBT), l0-ethyoxy-9-hydroxy-delta-6a-tetrahydrocannabinol, 8,9- dihydroxyl-delta-6a-tetrahydrocannabinol, cannabitriolvarin (CBTVE), dehydrocannabifuran (DCBF), cannabifuran (CBF), cannabichromanon (CBCN), cannabicitran (CBT), 10-oxo- delta-6a-tetrahydrocannabinol (OTHC), delta-9-cis-tetrahydrocannabinol (cis-THC), 3, 4,5,6- tetrahydro-7-hydroxy-alpha-alpha-2-trimethyl-9-n-propyl-2,6-methano-2H-l-benzoxocin-5- methanol (OH-iso-HHCV), cannabiripsol (CBR), trihydroxy-del ta-9-tetrahydrocannabinol (triOH-THC), CBGA-hydrocinnamic acid (3-[(2E)-3,7-dimethylocta-2,6-dien-l-yl]-2,4- dihydroxy-6-(2-phenylethyl)benzoic acid), CBG-hydrocinnamic acid (2-[(2E)-3 ,7- dimethyl octa-2,6-dien- 1 -yl]-5-(2-phenyl ethyl )benzene- 1 ,3 -diol), CBDA-hydrocinnamic acid (2,4-dihydroxy-3-[3-methyl-6-(prop-l-en-2-yl)cyclohex-2-en-l-yl]-6-(2- phenylethyl)benzoic acid), CBD-hydrocinnamic acid (2-[3-methyl-6-(prop-l-en-2- yl)cyclohex-2-en-l-yl]-5-(2-phenylethyl)benzene-l,3-diol), THCA-hydrocinnamic acid (1- hydroxy-6,6,9-trimethyl-3-(2-phenylethyl)-6H,6aH,7H,8H, l0aH-benzo[c]isochromene-2- carboxylic acid), THC-hydrocinnamic acid (6,6,9-trimethyl-3-(2-phenylethyl)- 6H,6aH,7H,8H,l0aH-benzo[c]isochromen-l-ol, perrottetinene), and derivatives of any of the foregoing. In some embodiments, the cannabinoid or cannabinoid derivative is produced in an amount of more than 100 mg/L culture medium. In some embodiments, the cannabinoid or cannabinoid derivative is produced in an amount of more than 50 mg/L culture medium.

[001044] In some embodiments, the cannabinoid produced with the methods or modified host cells of the present disclosure is cannabigerolic acid, cannabigerol, D⁹- tetrahydrocannabinolic acid, A⁹ -tetrahydrocannabinol, A⁸-tetrahydrocannabinolic acid, D⁸- tetrahydrocannabinol, cannabidiolic acid, cannabidiol, cannabichromenic acid,

cannabichromene, cannabinolic acid, cannabinol, cannabidivarinic acid, cannabidivarin, tetrahydrocannabivarinic acid, tetrahydrocannabivarin, cannabichromevarinic acid, cannabichromevarin, cannabigerovarinic acid, cannabigerovarin, cannabicyclolic acid, cannabicyclol, cannabielsoinic acid, cannabielsoin, cannabicitranic acid, or cannabicitran. In some embodiments, the cannabinoid is produced in an amount of more than 100 mg/L culture medium. In some embodiments, the cannabinoid is produced in an amount of more than 50 mg/L culture medium. [001045] Additional cannabinoid derivatives that can be produced with the methods or modified host cells of the present disclosure may also include, but are not limited to, 2- geranyl-5-pentyl-resorcylic acid, 2-geranyl-5-(4-pentynyl)-resorcylic acid, 2-geranyl-5- (trans-2-pentenyl)-resorcylic acid, 2-geranyl-5-(4-methylhexyl)-resorcylic acid, 2-geranyl-5- (5-hexynyl) resorcylic acid, 2-geranyl-5-(trans-2-hexenyl)-resorcylic acid, 2-geranyl-5-(5- hexenyl)-resorcylic acid, 2-geranyl-5-heptyl-resorcylic acid, 2-geranyl-5-(6-heptynoic)- resorcylic acid, 2-geranyl-5-octyl-resorcylic acid, 2-geranyl-5-(trans-2-octenyl)-resorcylic acid, 2-geranyl-5-nonyl-resorcylic acid, 2-geranyl-5-(trans-2-nonenyl) resorcylic acid, 2- geranyl-5-decyl-resorcylic acid, 2-geranyl-5-(4-phenylbutyl)-resorcylic acid, 2-geranyl-5-(5- phenylpentyl)-resorcylic acid, 2-geranyl-5-(6-phenylhexyl)-resorcylic acid, 2-geranyl-5-(7- phenylheptyl)-resorcylic acid, (6aR,10aR)-l -hydroxy-6, 6, 9-trimethyl-3-propyl-6a, 7,8, 10a- tetrahydro-6H-dibenzo[b,d]pyran-2-carboxylic acid, (6aR, l0aR)-l-hydroxy-6,6,9-trimethyl- 3-(4-methylhexyl)-6a,7,8,l0a-tetrahydro-6H-dibenzo[b,d]pyran-2-carboxylic acid,

(6aR,l0aR)-l-hydroxy-6,6,9-trimethyl-3-(5-hexenyl)-6a,7,8,l0a-tetrahydro-6H- dibenzo[b,d]pyran-2-carboxylic acid, (6aR, l0aR)-l-hydroxy-6,6,9-trimethyl-3-heptyl- 6a, 7, 8, l0a-tetrahydro-6H-dibenzo[b,d]pyran-2-carboxylic acid, (6aR, l0aR)-l -hydroxy- 6, 6, 9-trimethyl-3-(6-heptynyl)-6a, 7,8, 10a-tetrahydro-6H-dibenzo[b,d]pyran-2-carboxylic acid, 3-[(2E)-3,7-dimethylocta-2,6-dien-l-yl]-6-(hexan-2-yl)-2,4-dihydroxybenzoic acid, 3- [(2E)-3,7-dimethylocta-2,6-dien-l-yl]-2,4-dihydroxy-6-(2-methylpentyl)benzoic acid, 3- [(2E)-3,7-dimethylocta-2,6-dien-l-yl]-2,4-dihydroxy-6-(3-methylpentyl)benzoic acid, 3- [(2E)-3,7-dimethylocta-2,6-dien-l-yl]-2,4-dihydroxy-6-(4-methylpentyl)benzoic acid, 3- [(2E)-3,7-dimethylocta-2,6-dien-l-yl]-2,4-dihydroxy-6-[(lE)-pent-l-en-l-yl]benzoic acid, 3- [(2E)-3,7-dimethylocta-2,6-dien-l-yl]-2,4-dihydroxy-6-[(2E)-pent-2-en-l-yl]benzoic acid, 3- [(2E)-3,7-dimethylocta-2,6-dien-l-yl]-2,4-dihydroxy-6-[(2E)-pent-3-en-l-yl]benzoic acid, 3- [(2E)-3,7-dimethylocta-2,6-dien-l-yl]-2,4-dihydroxy-6-(pent-4-en-l-yl)benzoic acid, 3- [(2E)-3,7-dimethylocta-2,6-dien-l-yl]-2,4-dihydroxy-6-propylbenzoic acid, 3-[(2E)-3,7- dimethylocta-2,6-dien-l-yl]-2,4-dihydroxy-6-butylbenzoic acid, 3-[(2E)-3,7-dimethylocta- 2,6-dien-l-yl]-2,4-dihydroxy-6-hexylbenzoic acid, 3-[(2E)-3,7-dimethylocta-2,6-dien-l-yl]- 2,4-dihydroxy-6-heptylbenzoic acid, 3-[(2E)-3,7-dimethylocta-2,6-dien-l-yl]-2,4-dihydroxy- 6-octylbenzoic acid, 3-[(2E)-3,7-dimethylocta-2,6-dien-l-yl]-2,4-dihydroxy-6- nonanylbenzoic acid, 3-[(2E)-3,7-dimethylocta-2,6-dien-l-yl]-2,4-dihydroxy-6- decanylbenzoic acid, 3-[(2E)-3,7-dimethylocta-2,6-dien-l-yl]-2,4-dihydroxy-6- undecanylbenzoic acid, 6-(4-chlorobutyl)-3-[(2E)-3,7-dimethylocta-2,6-dien-l-yl]-2,4- dihydroxybenzoic acid, 3-[(2E)-3,7-dimethylocta-2,6-dien-l-yl]-2,4-dihydroxy-6-[4- (methylsulfanyl)butyl]benzoic acid, and others as listed in Bow, E. W. and Rimoldi, J. M., “The Structure-Function Relationships of Classical Cannabinoids: CB1/CB2 Modulation,” Perspectives in Medicinal Chemistry 2016:8 17-39 doi: 10.4137/PMC.S32171, incorporated by reference herein. In some embodiments, the cannabinoid derivative is produced in anamount of more than 100 mg/L culture medium. In some embodiments, the cannabinoid derivative is produced in an amount of more than 50 mg/L culture medium.

[001046] Additional cannabinoids and cannabinoid derivatives that can be produced with the methods or modified host cells of the present disclosure may also include, but are not limited to, 3-[(2E)-3,7-dimethylocta-2,6-dien-l-yl]-6-(hexan-2-yl)-2,4-dihydroxybenzoic acid, 3-[(2E)-3, 7-dimethyl octa-2,6-dien-l-yl]-2,4-dihydroxy-6-(3-methylpentyl)benzoic acid, 3-[(2E)-3, 7-dimethyl octa-2,6-dien-l-yl]-2,4-dihydroxy-6-(4-methylpentyl)benzoic acid, 3-[(2E)-3, 7-dimethyl octa-2,6-dien-l-yl]-2,4-dihydroxy-6-[(lE)-pent-l-en-l-yl]benzoic acid, 3-[(2E)-3, 7-dimethyl octa-2,6-dien-l-yl]-2,4-dihydroxy-6-[(2E)-pent-2-en-l-yl]benzoic acid, 3-[(2E)-3, 7-dimethyl octa-2,6-dien-l-yl]-2,4-dihydroxy-6-(pent-3-en-l-yl)benzoic acid, 3-[(2E)-3,7-dimethylocta-2,6-dien-l-yl]-2,4-dihydroxy-6-(pent-4-en-l-yl)benzoic acid, 3- [(2E)-3,7-dimethylocta-2,6-dien-l-yl]-2,4-dihydroxy-6-propylbenzoic acid, 3-[(2E)-3,7- dimethylocta-2,6-dien-l-yl]-2,4-dihydroxy-6-butylbenzoic acid, 3-[(2E)-3,7-dimethylocta- 2,6-dien-l-yl]-2,4-dihydroxy-6-hexylbenzoic acid, 3-[(2E)-3,7-dimethylocta-2,6-dien-l-yl]- 2,4-dihydroxy-6-heptylbenzoic acid, 6-(4-chlorobutyl)-3-[(2E)-3, 7-dimethyl octa-2,6-dien-l- yl]-2,4-dihydroxybenzoic acid, 3-[(2E)-3,7-dimethylocta-2,6-dien-l-yl]-2,4-dihydroxy-6-[4- (methylsulfanyl)butyl]benzoic acid, 3-[(2E)-3, 7-dimethyl octa-2,6-dien-l-yl]-2, 4-dihydroxy - 6-methylbenzoic acid, 2,4-dihydroxy-3-[(lR,6R)-3-methyl-6-(prop-l-en-2-yl)cyclohex-2- en-l-yl]-6-propylbenzoic acid, 6-butyl-2,4-dihydroxy-3-[(lR,6R)-3-methyl-6-(prop-l-en-2- yl)cyclohex-2-en-l-yl]benzoic acid, CBDA, CBD, CBGA, 6-hexyl-2,4-dihydroxy-3- [(lR,6R)-3-methyl-6-(prop-l-en-2-yl)cyclohex-2-en-l-yl]benzoic acid, THC, THCA, THCVA, CBDVA, (6aR, l0aR)-l-hydroxy-6,6,9-trimethyl-3-butyl-6a,7,8, 1 Oa-tetrahydro- 6H-dibenzo[b,d]pyran-2-carboxylic acid, (6aR, l0aR)-l-hydroxy-6,6,9-trimethyl-3-(3- methylpentyl)-6a,7,8, l0a-tetrahydro-6H-dibenzo[b,d]pyran-2-carboxylic acid, (6aR,l0aR)- 1 -hydroxy-6, 6, 9-trimethyl-3-(4-pentenyl)-6a, 7, 8, 10a-tetrahydro-6H-dibenzo[b,d]pyran-2- carboxylic acid, (6aR, 10aR)-l -hydroxy-6, 6, 9-trimethyl-3-hexyl-6a, 7,8,10a-tetrahydro-6H- dibenzo[b,d]pyran-2-carboxylic acid, (6aR, l0aR)-l-hydroxy-6,6,9-trimethyl-3-(5-hexynyl)- 6a,7,8,l0a-tetrahydro-6H-dibenzo[b,d]pyran-2-carboxylic acid, (E)-6-(but-3-yn-l-yl)-3-(3,7- dimethylocta-2,6-dien-l-yl)-2,4-dihydroxybenzoic acid, 6-((E)-but-l-en-l-yl)-3-((E)-3,7- dimethylocta-2,6-dien-l-yl)-2,4-dihydroxybenzoic acid, (E)-3-(3,7-dimethylocta-2,6-dien-l- yl)-2,4-dihydroxy-6-(pent-4-yn-l-yl)benzoic acid, 3-((E)-3,7-dimethylocta-2,6-dien-l-yl)-

2.4-dihydroxy-6-((E)-pent-l-en-l-yl)benzoic acid, (E)-3-(3,7-dimethylocta-2,6-dien-l-yl)-6- (hex-5-yn-l-yl)-2,4-dihydroxybenzoic acid, 3-((E)-3,7-dimethylocta-2,6-dien-l-yl)-6-((E)- hept-l-en-l-yl)-2,4-dihydroxybenzoic acid, (E)-3-(3,7-dimethylocta-2,6-dien-l-yl)-2,4- dihydroxy-6-octylbenzoic acid, 3-((E)-3,7-dimethylocta-2,6-dien-l-yl)-2,4-dihydroxy-6- ((E)-oct-l-en-l-yl)benzoic acid, (E)-3-(3, 7-dimethyl octa-2,6-di en-l-yl)-2, 4-dihydroxy-6- nonylbenzoic acid, 3-[(2E)-3,7-dimethylocta-2,6-dien-l-yl]-2,4-dihydroxy-6- undecanylbenzoic acid, (E)-3-(3,7-dimethylocta-2,6-dien-l-yl)-2,4-dihydroxy-6-(3- phenylpropyl)benzoic acid, (E)-3-(3,7-dimethylocta-2,6-dien-l-yl)-2,4-dihydroxy-6-(4- phenylbutyl)benzoic acid, (E)-3-(3,7-dimethylocta-2,6-dien-l-yl)-2,4-dihydroxy-6-(5- phenylpentyl)benzoic acid, and (E)-3-(3,7-dimethylocta-2,6-dien-l-yl)-2,4-dihydroxy-6-(6- phenylhexyl)benzoic acid. In some embodiments, the cannabinoid or cannabinoid derivative is produced in an amount of more than 100 mg/L culture medium. In some embodiments, the cannabinoid or cannabinoid derivative is produced in an amount of more than 50 mg/L culture medium.

[001047] Additional cannabinoids and cannabinoid derivatives that can be produced with the methods or modified host cells of the present disclosure may also include, but are not limited to, 3-[(2E)-3,7-dimethylocta-2,6-dien-l-yl]-6-(hexan-2-yl)-2,4-dihydroxybenzoic acid, 3-[(2E)-3, 7-dimethyl octa-2,6-dien-l-yl]-2,4-dihydroxy-6-(2-methylpentyl)benzoic acid, 3-[(2E)-3, 7-dimethyl octa-2,6-dien-l-yl]-2,4-dihydroxy-6-(3-methylpentyl)benzoic acid, 3-[(2E)-3, 7-dimethyl octa-2,6-dien-l-yl]-2,4-dihydroxy-6-(4-methylpentyl)benzoic acid, 3-[(2E)-3, 7-dimethyl octa-2,6-dien-l-yl]-2,4-dihydroxy-6-[(lE)-pent-l-en-l-yl]benzoic acid, 3-[(2E)-3, 7-dimethyl octa-2,6-dien-l-yl]-2,4-dihydroxy-6-[(2E)-pent-2-en-l-yl]benzoic acid, 3-[(2E)-3, 7-dimethyl octa-2,6-dien-l-yl]-2,4-dihydroxy-6-[(2E)-pent-3-en-l-yl]benzoic acid, 3-[(2E)-3, 7-dimethyl octa-2,6-dien-l-yl]-2,4-dihydroxy-6-(pent-4-en-l-yl)benzoic acid, 3-[(2E)-3,7-dimethylocta-2,6-dien-l-yl]-2,4-dihydroxy-6-propylbenzoic acid, 3-[(2E)-3,7- dimethylocta-2,6-dien-l-yl]-2,4-dihydroxy-6-butylbenzoic acid, 3-[(2E)-3,7-dimethylocta- 2,6-dien-l-yl]-2,4-dihydroxy-6-hexylbenzoic acid, 3-[(2E)-3,7-dimethylocta-2,6-dien-l-yl]-

2.4-dihydroxy-6-heptylbenzoic acid, 3-[(2E)-3,7-dimethylocta-2,6-dien-l-yl]-2, 4-dihydroxy - 6-octylbenzoic acid, 3-[(2E)-3,7-dimethylocta-2,6-dien-l-yl]-2,4-dihydroxy-6- nonanylbenzoic acid, 3-[(2E)-3,7-dimethylocta-2,6-dien-l-yl]-2,4-dihydroxy-6- decanylbenzoic acid, 3-[(2E)-3,7-dimethylocta-2,6-dien-l-yl]-2,4-dihydroxy-6- undecanylbenzoic acid, 6-(4-chlorobutyl)-3-[(2E)-3,7-dimethylocta-2,6-dien-l-yl]-2,4- dihydroxybenzoic acid, 3-[(2E)-3,7-dimethylocta-2,6-dien-l-yl]-2,4-dihydroxy-6-[4- (methylsulfanyl)butyl]benzoic acid, and 3-[(2E)-3,7-dimethylocta-2,6-dien-l-yl]-2,4- dihydroxy-6-methylbenzoic acid. In some embodiments, the cannabinoid or cannabinoid derivative is produced in an amount of more than 100 mg/L culture medium. In some embodiments, the cannabinoid or cannabinoid derivative is produced in an amount of more than 50 mg/L culture medium.

[001048] Additional cannabinoids and cannabinoid derivatives that can be produced with the methods or modified host cells of the present disclosure may also include, but are not limited to, (rR,2'R)-4-(hexan-2-yl)-5'-methyl-2'-(prop-l-en-2-yl)-l',2',3',4'-tetrahydro- [ 1 , l'-biphenyl]-2,6-diol, (rR,2'R)-4-hexyl-5'-methyl-2'-(prop- 1 -en-2-yl)- 1 ',2',3',4'- tetrahydro-[l,l'-biphenyl]-2,6-diol, (l'R,2'R)-5'-methyl-4-(3-methylpentyl)-2'-(prop-l-en-2- yl)-r,2',3',4'-tetrahydro-[l, r-biphenyl]-2,6-diol, (rR,2'R)-4-(4-chlorobutyl)-5'-methyl-2'- (prop-l-en-2-yl)-r,2',3',4'-tetrahydro-[l,l'-biphenyl]-2,6-diol, (l'R,2'R)-5'-methyl-4-(4- methylpentyl)-2'-(prop-l-en-2-yl)-r,2',3',4'-tetrahydro-[l, r-biphenyl]-2,6-diol, (l'R,2'R)-5'- methyl-4-(4-(methylthio)butyl)-2'-(prop-l-en-2-yl)-r,2',3',4'-tetrahydro-[l, l'-biphenyl]-2,6- diol, (l'R,2'R)-5'-methyl-4-((E)-pent-l-en-l-yl)-2'-(prop-l-en-2-yl)-l',2',3',4'-tetrahydro- [l,l'-biphenyl]-2,6-diol, (l'R,2'R)-5'-methyl-4-((E)-pent-3-en-l-yl)-2'-(prop-l-en-2-yl)- l',2',3',4'-tetrahydro-[l,r-biphenyl]-2,6-diol, (rR,2'R)-5'-methyl-4-((E)-pent-2-en-l-yl)-2'- (prop-l-en-2-yl)-r,2',3',4'-tetrahydro-[l,l'-biphenyl]-2,6-diol, (l'R,2'R)-4-(but-3-yn-l-yl)-5'- methyl-2'-(prop-l-en-2-yl)-l',2',3',4'-tetrahydro-[l,r-biphenyl]-2,6-diol, (ER,2'R)-4-((E)- but- 1 -en- 1 -yl)-5'-methyl-2'-(prop- 1 -en-2-yl)- 1 ',2', 3 ',4'-tetrahydro-[l , 1 '-biphenyl]-2,6-diol, (rR,2'R)-5'-methyl-4-(pent-4-yn-l-yl)-2'-(prop-l-en-2-yl)-l',2',3',4'-tetrahydro-[l, r- biphenyl]-2,6-diol, (l^lR,2^lR)-5^l-methyl-2'-(prop-l-en-2-yl)-4-undecyl-l^l,2',3',4'-tetrahydro- [l,l'-biphenyl]-2,6-diol, (lR,2R)-4-(hex-5-yn-l-yl)-5'-methyl-2'-(prop-l-en-2-yl)-l',2',3',4'- tetrahydro-[ 1 , 1 '-biphenyl]-2,6-diol, (l'R,2'R)-4-((E)-hept- 1 -en- 1 -yl)-5'-methyl-2'-(prop- 1 -en- 2-yl)-r,2',3',4'-tetrahydro-[l,l'-biphenyl]-2,6-diol, (l'R,2'R)-5'-methyl-4-octyl-2'-(prop-l-en- 2-yl)- 1',2',3 ',4'-tetrahy dro-[ 1 , 1 '-biphenyl]-2,6-diol, ( 1 'R,2'R)-5'-methyl-4-((E)-oct- 1 -en- 1 -yl)- 2'-(prop-l-en-2-yl)-l',2',3',4'-tetrahydro-[l,l'-biphenyl]-2,6-diol, (l'R,2'R)-5'-methyl-4- nonyl-2'-(prop-l-en-2-yl)-r,2',3',4'-tetrahydro-[l,r-biphenyl]-2,6-diol, (l'R,2'R)-5'-methyl- 4-(3-phenylpropyl)-2'-(prop-l-en-2-yl)-l',2',3',4'-tetrahydro-[l, l'-biphenyl]-2,6-diol, (rR,2'R)-5'-methyl-4-(4-phenylbutyl)-2'-(prop-l-en-2-yl)-l',2',3',4'-tetrahydro-[l,r- biphenyl]-2,6-diol, (rR,2'R)-5^l-methyl-4-(5-phenylpentyl)-2'-(prop-l-en-2-yl)-l',2',3',4'- tetrahydro-[ 1 , 1 '-biphenyl]-2,6-diol, (l'R,2'R)-5'-methyl-4-(6-phenylhexyl)-2'-(prop- 1 -en-2- yl)-l',2',3',4'-tetrahydro-[l, r-biphenyl]-2,6-diol, (l'R,2'R)-5'-methyl-4-(2-methylpentyl)-2'- (prop-l-en-2-yl)-r,2',3',4'-tetrahydro-[l,l'-biphenyl]-2,6-diol, (l'R,2'R)-4-isopropyl-5'- methyl-2'-(prop-l-en-2-yl)- ,2',3',4'-tetrahydro-[l,l'-biphenyl]-2,6-diol, (l'R,2'R)-4-decyl- 5'-methyl-2'-(prop-l-en-2-yl)-l',2',3',4'-tetrahydro-[l,r-biphenyl]-2,6-diol, (l'R,2R)-5'- methyl-2'-(prop-l-en-2-yl)-4-tridecyl-l',2',3',4'-tetrahydro-[l,r-biphenyl]-2,6-diol, (E)-3- ((rR,2'R)-2,6-dihydroxy-5'-methyl-2'-(prop-l-en-2-yl)-r, 2', 3', 4'-tetrahydro-[ 1, 1 '-biphenyl]- 4-yl)acrylic acid, (Z)-3-((rR,2'R)-2,6-dihydroxy-5'-methyl-2'-(prop-l-en-2-yl)-r,2',3',4'- tetrahydro-[l,l'-biphenyl]-4-yl)acrylic acid, 7-((rR,2'R)-2,6-dihydroxy-5'-methyl-2'-(prop- l-en-2-yl)-l',2',3',4'-tetrahydro-[l, r-biphenyl]-4-yl)heptanoic acid, 8-((l'R,2'R)-2,6- dihydroxy-5'-methyl-2'-(prop-l-en-2-yl)-r,2',3',4'-tetrahydro-[l,r-biphenyl]-4-yl)octanoic acid, 9-((rR,2'R)-2,6-dihydroxy-5'-methyl-2'-(prop-l-en-2-yl)-r,2',3',4^,-tetrahydro-[l,r- biphenyl]-4-yl)nonanoic acid, 1 l-((l'R,2'R)-2,6-dihydrc>xy-5'-methyl-2'-(prop-l-en-2-yl)- l',2',3',4'-tetrahydro-[l,r-biphenyl]-4-yl)undecanoic acid, (l"R,2"R)-3',5'-dihydroxy-5"- methyl-2"-(prop- 1 -en-2-yl)- 1 ",2",3",4"-tetrahydro-[ 1 , 1 ' :4', 1 "-terphenyl]-2-carboxylic acid, (rⁱR,2^l'R)-3',5'-dihydroxy-5"-methyl-2"-(prop-l-en-2-yl)-l",2",3^l',4"-tetrahydro-[l,r:4',l"- terphenyl]-3-carboxylic acid, (l"R,2"R)-3',5'-dihydroxy-5"-methyl-2"-(prop-l-en-2-yl)- l",2",3",4"-tetrahydro-[l,l':4',l"-terphenyl]-4-carboxylic acid, (l"R,2"R)-3',5'-dihydroxy-5"- methyl-2"-(prop-l-en-2-yl)-l",2",3",4^l'-tetrahydro-[l,l':4',rⁱ-terphenyl]-3,5-dicarboxylic acid, (rR,2'R)-4-(4-hydroxybutyl)-5'-methyl-2'-(prop-l-en-2-yl)-r,2',3',4'-tetrahydro-[l,r- biphenyl]-2,6-diol, (rR,2'R)-4-(4-aminobutyl)-5'-methyl-2'-(prop-l-en-2-yl)-r,2',3',4'- tetrahydro-[l,l'-biphenyl]-2,6-diol, 5-((rR,2'R)-2,6-dihydroxy-5'-methyl-2'-(prop-l-en-2- y 1 )- 1 ',2',3',4'-tetrahydro-[ l , 1 '-bipheriyl]-4-yl)pemanenilrile, (rR,2'R)-5'-methyl-4-(3- methylhexan-2-yl)-2'-(prop-l-en-2-yl)-r,2',3',4'-tetrahydro-[l,r-biphenyl]-2,6-diol, (rR,2'R)-5'-methyl-2'-(prop-l-en-2-yl)-4-propyl-r,2',3',4'-tetrahydro-[l,r-biphenyl]-2,6- diol, (l'R,2'R)-4-butyl-5'-methyl-2'-(prop-l-en-2-yl)-l',2',3',4'-tetrahydro-[l,r-biphenyl]- 2,6-diol, (rR,2'R)-5'-methyl-4-pentyl-2'-(prop-l-en-2-yl)-l',2',3',4'-tetrahydro-[l,l'- biphenyl]-2,6-diol, (rR,2'R)-4-heptyl-5'-methyl-2'-(prop-l-en-2-yl)-r,2',3^l,4'-tetrahydro- [l,l'-biphenyl]-2,6-diol, (rR,2'R)-5'-methyl-4-(pent-4-en-l-yl)-2'-(prop-l-en-2-yl)-r,2',3',4'- tetrahydro-[l,l'-biphenyl]-2,6-diol, 3-((l'R,2'R)-2,6-dihydroxy-5'-methyl-2'-(prop-l-en-2- yl)-r,2',3',4'-tetrahydro-[l, r-biphenyl]-4-yl)propanoic acid, (l'R,2'R)-4,5'-dimethyl-2'- (prop-l-en-2-yl)-r,2',3',4'-tetrahydro-[l,r-biphenyl]-2,6-diol, 2-((rR,2'R)-2,6-dihydroxy-5'- methyl-2'-(prop-l-en-2-yl)-r,2',3',4'-tetrahydro-[l,r-biphenyl]-4-yl)acetic acid, 4- ((rR,2'R)-2,6-dihydroxy-5'-methyl-2'-(prop-l-en-2-yl)-r,2',3',4'-tetrahydro-[l, l'-biphenyl]- 4-yl)butanoic acid, (rR,2'R)-2,6-dihydroxy-5'-methyl-2'-(prop-l-en-2-yl)-r,2',3',4'- tetrahydro-[ 1 , 1 '-biphenyl]-4-carboxylic acid, 5-(( 1 'R,2'R)-2,6-dihydroxy-5'-methyl-2'-(prop- l-en-2-yl)-l',2',3',4'-tetrahydro-[l, r-biphenyl]-4-yl)pentanoic acid, and 6-((l'R,2'R)-2,6- dihydroxy-5'-methyl-2'-(prop-l-en-2-yl)- ,2',3',4'-tetrahydro-[l, -biphenyl]-4-yl)hexanoic acid. In some embodiments, the cannabinoid or cannabinoid derivative is produced in an amount of more than 100 mg/L culture medium. In some embodiments, the cannabinoid or cannabinoid derivative is produced in an amount of more than 50 mg/L culture medium.

[001049] In some embodiments, the cannabinoids or cannabinoid derivatives that can be produced with the methods or modified host cells of the present disclosure may also include compounds of Formula (I):

or pharmaceutically acceptable salts thereof, wherein,

R¹ is a Ci-Cisalkyl group substituted with R^a or R¹ is a Ci-C ixalkenyl group substituted with R^a, wherein R^a is -S-Ci-C6alkyl or -S-C2-C6alkenyl, or R¹ is selected from the group consisting of:

In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is produced in an amount of more than 100 mg/L culture medium. In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is produced in an amount of more than 50 mg/L culture medium.

[001050] In some embodiments the compound of Formula (I), or a pharmaceutically acceptable salt thereof, may include where R¹ is a C i-Cixalkyl group substituted with R^a or R¹ is a C2-Ci8alkenyl group substituted with R^a, wherein R^a is S-Ci-C6alkyl or S-C2- Csalkenyl, or R¹ is selected from the group consisting of:

[001051] In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, may include where R¹ is selected from the group consisting of:

[001052] In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, may include where R¹ is a Ci-Ci8alkyl group substituted with R^a or R¹ is a C2-Ci8alkenyl group substituted with R^a, wherein R^a is -S-Ci-C6alkyl or -S-C2- C6alkenyl.

[001053] In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, may include where R¹ is selected from the group consisting of:

[001054] In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is selected from the group consisting of:

444

445

[001055] In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is selected from the group consisting of:

[001056] In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is selected from the group consisting of:

[001057] A cannabinoid derivative may also refer to a compound lacking one or more chemical moieties found in naturally-occurring cannabinoids, yet retains the core structural features (e.g., cyclic core) of a naturally-occurring cannabinoid. Such chemical moieties may include, but are not limited to, methyl, alkyl, alkenyl, methoxy, alkoxy, acetyl, carboxyl, carbonyl, oxo, ester, hydroxyl, and the like. In some embodiments, a cannabinoid derivative may also comprise one or more of any of the functional and/or reactive groups described herein. Functional and reactive groups may be unsubstituted or substituted with one or more functional or reactive groups.

[001058] A cannabinoid derivative may be a cannabinoid substituted with or comprising one or more functional and/or reactive groups. Functional groups may include, but are not limited to, azido, halo (e.g., chloride, bromide, iodide, fluorine), methyl, alkyl, alkynyl, alkenyl, methoxy, alkoxy, acetyl, amino, carboxyl, carbonyl, oxo, ester, hydroxyl, thio (e g., thiol), cyano, aryl, heteroaryl, cycloalkyl, cycloalkenyl, cycloalkylalkenyl, cycloalkylalkynyl, cycloalkenylalkyl, cycloalkenylalkenyl, cycloalkenylalkynyl,

heterocyclylalkenyl, heterocyclylalkynyl, heteroarylalkenyl, heteroaryl alkynyl, arylalkenyl, arylalkynyl, spirocyclyl, heterospirocyclyl, heterocyclyl, thioalkyl (or alkylthio), arylthio, heteroarylthio, sulfone, sulfonyl, sulfoxide, amido, alkylamino, dialkylamino, arylamino, alkyl aryl amino, diarylamino, N-oxide, imide, enamine, imine, oxime, hydrazone, nitrile, aralkyl, cycloalkylalkyl, haloalkyl, heterocyclylalkyl, heteroarylalkyl, nitro, thioxo, and the like. Suitable reactive groups may include, but are not necessarily limited to, azide, carboxyl, carbonyl, amine (e.g., alkyl amine (e.g., lower alkyl amine), aryl amine), halide, ester (e.g., alkyl ester (e.g., lower alkyl ester, benzyl ester), aryl ester, substituted aryl ester), cyano, thioester, thioether, sulfonyl halide, alcohol, thiol, succinimidyl ester, isothiocyanate, iodoacetamide, maleimide, hydrazine, alkynyl, alkenyl, acetyl, and the like. In some embodiments, the reactive group is selected from a carboxyl, a carbonyl, an amine, an ester, a thioester, a thioether, a sulfonyl halide, an alcohol, a thiol, an alkyne, alkene, an azide, a succinimidyl ester, an isothiocyanate, an iodoacetamide, a maleimide, and a hydrazine. Functional and reactive groups may be unsubstituted or substituted with one or more functional or reactive groups.

[001059] “Alkyl” may refer to a straight or branched chain saturated hydrocarbon. For example, Ci-C6alkyl groups contain 1 to 6 carbon atoms. Examples of a Ci-C6alkyl group include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, isopropyl, isobutyl, sec- butyl and tert- butyl, isopentyl, and neopentyl.

[001060] “Alkenyl” may include an unbranched (i.e., straight) or branched

hydrocarbon chain containing 2-12 carbon atoms. The“alkenyl” group contains at least one double bond. The double bond of an alkenyl group can be unconjugated or conjugated to another unsaturated group. Examples of alkenyl groups may include, but are not limited to, ethylenyl, vinyl, allyl, butenyl, pentenyl, hexenyl, butadienyl, pentadienyl, hexadienyl, 2- ethylhexenyl, 2-propyl-2-butenyl, 4-(2-methyl-3-butene)-pentenyl and the like. [001061] Compounds disclosed herein, such as cannabinoids and cannabinoid derivatives, may be substituted with one or more substituents, such as those illustrated generally herein, or as exemplified by particular classes, subclasses, and species of the present disclosure. In general, the term“substituted” refers to the replacement of a hydrogen atom in a given structure with a specified substituent. Combinations of substituents envisioned by the present disclosure are typically those that result in the formation of stable or chemically feasible compounds.

[001062] As used herein, the term“unsubstituted” may mean that the specified group bears no substituents beyond the moiety recited (e.g., where valency satisfied by hydrogen).

[001063] A reactive group may facilitate covalent attachment of a molecule of interest. Suitable molecules of interest may include, but are not limited to, a detectable label; imaging agents; a toxin (including cytotoxins); a linker; a peptide; a drug (e.g., small molecule drugs); a member of a specific binding pair; an epitope tag; ligands for binding by a target receptor; tags to aid in purification; molecules that increase solubility; and the like. A linker may be a peptide linker or a non-peptide linker.

[001064] In some embodiments, a cannabinoid derivative substituted with an azide may be reacted with a compound comprising an alkyne group via“click chemistry” to generate a product comprising a heterocycle, also known as an azide-alkyne cycloaddition. In some embodiments, a cannabinoid derivative substituted with an alkyne may be reacted with a compound comprising an azide group via click chemistry to generate a product comprising a heterocycle.

[001065] Additional molecules of interest that may be desirable for attachment to a cannabinoid derivative may include, but are not necessarily limited to, detectable labels (e.g., spin labels, fluorescence resonance energy transfer (FRET)-type dyes, e.g., for studying structure of biomolecules in vivo); small molecule drugs; cytotoxic molecules (e.g., drugs); imaging agents; ligands for binding by a target receptor; tags to aid in purification by, for example, affinity chromatography (e.g., attachment of a FLAG epitope); molecules that increase solubility (e.g., poly(ethylene glycol)); molecules that enhance bioavailability; molecules that increase in vivo half-life; molecules that target to a particular cell type (e.g., an antibody specific for an epitope on a target cell); molecules that target to a particular tissue; molecules that provide for crossing the blood-brain barrier; and molecules to facilitate selective attachment to a surface, and the like.

[001066] In some embodiments, a molecule of interest comprises an imaging agent. Suitable imaging agents may include positive contrast agents and negative contrast agents. Suitable positive contrast agents may include, but are not limited to, gadolinium

tetraazacyclododecanetetraacetic acid (Gd-DOTA); gadolinium- diethylenetriaminepentaacetic acid (Gd-DTPA); gadolinium-1, 4, 7-tris(carbonylmethyl)-l0- (2'-hydroxypropyl)-l,4,7,l0-tetraazacyclododecane (Gd-HP-D03A); Manganese(II)- dipyridoxal diphosphate (Mn-DPDP); Gd-diethylenetriaminepentaacetate-bis(methylamide) (Gd-DTPA-BMA); and the like. Suitable negative contrast agents may include, but are not limited to, a superparamagnetic iron oxide (SPIO) imaging agent; and a perfluorocarbon, where suitable perfluorocarbons may include, but are not limited to, fluoroheptanes, fluorocycloheptanes, fluoromethyl cycloheptanes, fluorohexanes, fluorocyclohexanes, fluoropentanes, fluorocyclopentanes, fluoromethylcyclopentanes,

fluorodimethylcyclopentanes, fluoromethyl cyclobutanes, fluorodimethylcyclobutanes, fluorotrimethylcyclobutanes, fluorobutanes, fluorocyclobutanse, fluoropropanes, fluoroethers, fluoropolyethers, fluorotriethylamines, perfluorohexanes, perfluoropentanes, perfluorobutanes, perfluoropropanes, sulfur hexafluoride, and the like.

[001067] Additional cannabinoid derivatives that can be produced with a method or modified host cell of the present disclosure may include derivatives that have been modified via organic synthesis or an enzymatic route to modify drug metabolism and

pharmacokinetics (e.g., solubility, bioavailability, absorption, distribution, plasma half-life and metabolic clearance). Modification examples may include, but are not limited to, halogenation, acetylation, and methylation.

[001068] The cannabinoids or cannabinoid derivatives described herein further include all pharmaceutically acceptable isotopically labeled cannabinoids or cannabinoid derivatives. An“isotopically-” or“radio-labeled” compound is a compound where one or more atoms are replaced or substituted by an atom having an atomic mass or mass number different from the atomic mass or mass number typically found in nature (i.e., naturally occurring). For example, in some embodiments, in the cannabinoids or cannabinoid derivatives described herein, hydrogen atoms are replaced or substituted by one or more deuterium or tritium. Certain isotopically labeled cannabinoids or cannabinoid derivatives of this disclosure, for example, those incorporating a radioactive isotope, are useful in drug and/or substrate tissue distribution studies. The radioactive isotopes tritium, i.e., ³H, and carbon 14, i.e., ¹⁴C, are particularly useful for this purpose in view of their ease of incorporation and ready means of detection. Substitution with heavier isotopes such as deuterium, i.e., ²H, may afford certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements, and hence may be preferred in some circumstances. Suitable isotopes that may be incorporated in cannabinoids or cannabinoid derivatives described herein include but are not limited to ²H (also written as D for deuterium), ¾ (also written as T for tritium), ^UC, ¹³C, ¹⁴C, ¹³N, ¹⁵N, ¹⁵0, ¹⁷0, ¹⁸0, ¹⁸F, ³⁵S, ³⁶C1 , ⁸²Br, ⁷⁵Br, ⁷⁶Br, ⁷⁷Br, ¹²³I, ¹²⁴I, ¹²⁵I, and ¹³¹I. Substitution with positron emitting isotopes, such as ¹ 'C, ¹⁸F, ¹⁵0, and ¹³N, can be useful in Positron Emission Topography (PET) studies.

[001069] The methods of bioproduction disclosed herein enable synthesis of cannabinoids or cannabinoid derivatives with defined stereochemistries, which is challenging to do using chemical synthesis. Cannabinoids or cannabinoid derivatives disclosed herein may be enantiomers or disastereomers. The term“enantiomers” may refer to a pair of stereoisomers which are non-superimposable mirror images of one another. In some embodiments the cannabinoids or cannabinoid derivatives may be the (ri)-enantiomer. In some embodiments the cannabinoids or cannabinoid derivatives may be the ( R )- enantiomer. In some embodiments, the cannabinoids or cannabinoid derivatives may be the (+) or (-) enantiomers. The term“diastereomers” may refer to the set of stereoisomers which cannot be made superimposable by rotation around single bonds. For example, cis- and trans- double bonds, endo- and exo- substitution on bicyclic ring systems, and compounds containing multiple stereogenic centers with different relative configurations may be considered to be diastereomers. The term“diastereomer” may refer to any member of this set of compounds. Cannabinoids or cannabinoid derivatives disclosed herein may include a double bond or a fused ring. In certain such embodiments, the double bond or fused ring may be cis or trans, unless the configuration is specifically defined. If the cannabinoid or cannabinoid derivative contains a double bond, the substituent may be in the E or Z configuration, unless the configuration is specifically defined.

[001070] In some embodiments when the cannabinoid or cannabinoid derivative is recovered from a cell lysate; from a culture medium; from a modified host cell; from both the cell lysate and the culture medium; from both the modified host cell and the culture medium; from the cell lysate, the modified host cell, and the culture medium; or from a cell- free reaction mixture comprising one or more polypeptides disclosed herein, the recovered cannabinoid or cannabinoid derivative is in the form of a salt. In certain such embodiments, the salt is a pharmaceutically acceptable salt. In some embodiments, the salt of the recovered cannabinoid or cannabinoid derivative is then purified as disclosed herein.

[001071] The disclosure includes pharmaceutically acceptable salts of the cannabinoids or cannabinoid derivatives described herein. “Pharmaceutically acceptable salts” may refer to those salts which retain the biological effectiveness and properties of the free bases, which are not biologically or otherwise undesirable. Representative pharmaceutically acceptable salts include, but are not limited to, e.g., water-soluble and water-insoluble salts, such as the acetate, amsonate (4, 4-diaminostilbene-2, 2-disulfonate), benzenesulfonate, benzonate, bicarbonate, bisulfate, bitartrate, borate, bromide, butyrate, calcium, calcium edetate, camsylate, carbonate, chloride, citrate, clavulariate, dihydrochloride, edetate, edisylate, estolate, esylate, fiunarate, gluceptate, gluconate, glutamate, glycollylarsanilate,

hexafluorophosphate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroxynaphthoate, iodide, sethionate, lactate, lactobionate, laurate, magnesium, malate, maleate, mandelate, mesylate, methylbromide, methylnitrate, methylsulfate, mucate, napsylate, nitrate, N-methylglucamine ammonium salt, 3-hydroxy-2-naphthoate, oleate, oxalate, palmitate, pamoate (l,l-methene-bis-2-hydroxy-3-naphthoate, einbonate), pantothenate, phosphate/diphosphate, picrate, polygalacturonate, propionate, p- toluenesulfonate, salicylate, stearate, subacetate, succinate, sulfate, sulfosalicylate, suramate, tannate, tartrate, teoclate, tosylate, triethiodide, and valerate salts.

[001072] “Pharmaceutically acceptable salt” also includes both acid and base addition salts. “Pharmaceutically acceptable acid addition salt” may refer to those salts which retain the biological effectiveness and properties of the free bases, which are not biologically or otherwise undesirable, and which are formed with inorganic acids such as, but are not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like, and organic acids such as, but not limited to, acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, camphoric acid, camphor- lO-sulfonic acid, capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecyl sulfuric acid, ethane- l,2-disulfonic acid, ethanesulfonic acid, 2- hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid, gluconic acid, glucuronic acid, glutamic acid, glutaric acid, 2-oxo- glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, mucic acid, naphthalene-1, 5-disulfonic acid, naphthalene-2-sulfonic acid, l-hydroxy-2-naphthoic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, pyroglutamic acid, pyruvic acid, salicylic acid, 4- aminosalicylic acid, sebacic acid, stearic acid, succinic acid, tartaric acid, thiocyanic acid, p- toluenesulfonic acid, trifluoroacetic acid, undecylenic acid, and the like. [001073] “Pharmaceutically acceptable base addition salt” may refer to those salts which retain the biological effectiveness and properties of the free acids, which are not biologically or otherwise undesirable. These salts are prepared from addition of an inorganic base or an organic base to the free acid. Salts derived from inorganic bases include, but are not limited to, the sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts and the like. For example, inorganic salts include, but are not limited to, ammonium, sodium, potassium, calcium, and magnesium salts. Salts derived from organic bases include, but are not limited to, salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, diethanolamine, ethanolamine, deanol, 2- dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, benethamine, benzathine, ethylenediamine, glucosamine, methylglucamine, theobromine, triethanolamine, tromethamine, purines, piperazine, piperidine, N-ethylpiperidine, polyamine resins and the like.

Methods of Using Host Cells to Generate Cannabinoids or Cannabinoid Derivatives

[001074] The disclosure provides methods of producing a cannabinoid or a cannabinoid derivative, such as those described herein, the method comprising culturing a modified host cell of the disclosure in a culture medium. In certain such embodiments, the method comprises recovering the produced cannabinoid or cannabinoid derivative. In certain such embodiments, the produced cannabinoid or cannabinoid derivative is then purified as disclosed herein.

[001075] In some embodiments, culturing of the modified host cells of the disclosure in a culture medium provides for synthesis of a cannabinoid or a cannabinoid derivative, such as those described herein, in an increased amount compared to an unmodified host cell cultured under similar conditions.

[001076] The disclosure provides methods of producing a cannabinoid or a cannabinoid derivative, such as those described herein, the method comprising: culturing a modified host cell of the disclosure in a culture medium comprising a carboxylic acid. In certain such embodiments, the method comprises recovering the produced cannabinoid or cannabinoid derivative. In certain such embodiments, the produced cannabinoid or cannabinoid derivative is then purified as disclosed herein. [001077] In some embodiments, the cannabinoid or cannabinoid derivative is recovered from a cell lysate; from a culture medium; from a modified host cell; from both the cell lysate and the culture medium; from both the modified host cell and the culture medium; or from the cell lysate, the modified host cell, and the culture medium. In certain such embodiments, the recovered cannabinoid or cannabinoid derivative is then purified as disclosed herein. In some embodiments when the cannabinoid or cannabinoid derivative is recovered from the cell lysate; from the culture medium; from the modified host cell; from both the cell lysate and the culture medium; from both the modified host cell and the culture medium; or from the cell lysate, the modified host cell, and the culture medium, the recovered cannabinoid or cannabinoid derivative is in the form of a salt. In certain such embodiments, the salt is a pharmaceutically acceptable salt. In some embodiments, the salt of the recovered cannabinoid or cannabinoid derivative is then purified as disclosed herein.

[001078] In some embodiments, the modified host cell of the present disclosure is cultured in a culture medium comprising a carboxylic acid. In some embodiments, the carboxylic acid may be substituted with or comprise one or more functional and/or reactive groups. Functional groups may include, but are not limited to, azido, halo (e.g., chloride, bromide, iodide, fluorine), methyl, alkyl, alkynyl, alkenyl, methoxy, alkoxy, acetyl, amino, carboxyl, carbonyl, oxo, ester, hydroxyl, thio (e.g., thiol), cyano, aryl, heteroaryl, cycloalkyl, cycloalkenyl, cycloalkylalkenyl, cycloalkylalkynyl, cycloalkenylalkyl, cycloalkenylalkenyl, cycloalkenylalkynyl, heterocyclylalkenyl, heterocyclylalkynyl, heteroarylalkenyl, heteroarylalkynyl, arylalkenyl, arylalkynyl, spirocyclyl, heterospirocyclyl, heterocyclyl, thioalkyl (or alkylthio), arylthio, heteroarylthio, sulfone, sulfonyl, sulfoxide, amido, alkylamino, dialkylamino, arylamino, alkylarylamino, diarylamino, N-oxide, imide, enamine, imine, oxime, hydrazone, nitrile, aralkyl, cycloalkylalkyl, haloalkyl,

heterocyclylalkyl, heteroarylalkyl, nitro, thioxo, and the like. Reactive groups may include, but are not necessarily limited to, azide, halogen, carboxyl, carbonyl, amine (e.g., alkyl amine (e.g., lower alkyl amine), aryl amine), ester (e.g., alkyl ester (e.g., lower alkyl ester, benzyl ester), aryl ester, substituted aryl ester), cyano, thioester, thioether, sulfonyl halide, alcohol, thiol, succinimidyl ester, isothiocyanate, iodoacetamide, maleimide, hydrazine, alkynyl, alkenyl, and the like. In some embodiments, the reactive group is selected from a carboxyl, a carbonyl, an amine, an ester, thioester, thioether, a sulfonyl halide, an alcohol, a thiol, a succinimidyl ester, an isothiocyanate, an iodoacetamide, a maleimide, an azide, an alkyne, an alkene, and a hydrazine. Functional and reactive groups may be unsubstituted or substituted with one or more functional or reactive groups. [001079] In some embodiments, the carboxylic acid is isotopically- or radio-labeled. In some embodiments, the carboxylic acid may be an enantiomer or disastereomer. In some embodiments the carboxylic acid may be the fS')-enantiomer. In some embodiments the carboxylic acid may be the (//)-enantiomer. In some embodiments, the carboxylic acid may be the (+) or (-) enantiomer. In some embodiments, the carboxylic acid may include a double bond or a fused ring. In certain such embodiments, the double bond or fused ring may be cis or trans, unless the configuration is specifically defined. If the carboxylic acid contains a double bond, the substituent may be in the E or Z configuration, unless the configuration is specifically defined.

[001080] In some embodiments, the carboxylic acid comprises a C=C group. In some embodiments, the carboxylic acid comprises an alkyne group. In some embodiments, the carboxylic acid comprises an N3 group. In some embodiments, the carboxylic acid comprises a halogen. In some embodiments, the carboxylic acid comprises a CN group. In some embodiments, the carboxylic acid comprises iodo. In some embodiments, the carboxylic acid comprises bromo. In some embodiments, the carboxylic acid comprises chloro. In some embodiments, the carboxylic acid comprises fluoro. In some embodiments, the carboxylic acid comprises a carbonyl. In some embodiments, the carboxylic acid comprises an acetyl.

In some embodiments, the carboxylic acid comprises an alkyl group. In some embodiments, the carboxylic acid comprises an aryl group.

[001081] Carboxylic acids may include, but are not limited to, unsubstituted or substituted C3-C18 fatty acids, C3-C18 carboxylic acids, Ci-Cis carboxylic acids, butyric acid, isobutyric acid, valeric acid, hexanoic acid, heptanoic acid, octanoic acid, nonanoic acid, decanoic acid, undecanoic acid, lauric acid, myristic acid, C 15-C18 fatty acids, C15-C18 carboxylic acids, fumaric acid, itaconic acid, malic acid, succinic acid, maleic acid, malonic acid, glutaric acid, glucaric acid, oxalic acid, adipic acid, pimelic acid, suberic acid, azelaic acid, sebacic acid, dodecanedioic acid, glutaconic acid, ortho-phthalic acid, isophthalic acid, terephthalic acid, citric acid, isocitric acid, aconitic acid, tricarballylic acid, and trimesic acid. Carboxylic acids may include unsubstituted or substituted Ci-Cis carboxylic acids. Carboxylic acids may include unsubstituted or substituted C3-C 18 carboxylic acids. Carboxylic acids may include unsubstituted or substituted C3-C12 carboxylic acids. Carboxylic acids may include unsubstituted or substituted C4-C10 carboxylic acids. In some embodiments, the carboxylic acid is an unsubstituted or substituted C4 carboxylic acid. In some embodiments, the carboxylic acid is an unsubstituted or substituted C5 carboxylic acid. In some embodiments, the carboxylic acid is an unsubstituted or substituted Cr, carboxylic acid. In some embodiments, the carboxylic acid is an unsubstituted or substituted Ci carboxylic acid. In some embodiments, the carboxylic acid is an unsubstituted or substituted Cx carboxylic acid. In some embodiments, the carboxylic acid is an unsubstituted or substituted C9 carboxylic acid. In some embodiments, the carboxylic acid is an unsubstituted or substituted C10 carboxylic acid. In some embodiments, the carboxylic acid is unsubstituted or substituted butyric acid. In some embodiments, carboxylic acid is unsubstituted or substituted valeric acid. In some embodiments, the carboxylic acid is unsubstituted or substituted hexanoic acid. In some embodiments, the carboxylic acid is unsubstituted or substituted heptanoic acid. In some embodiments, the carboxylic acid is unsubstituted or substituted octanoic acid. In some embodiments, the carboxylic acid is unsubstituted or substituted nonanoic acid. In some embodiments, the carboxylic acid is unsubstituted or substituted decanoic acid.

[001082] Carboxylic acids may include, but are not limited to, 2-methylhexanoic acid, 3-methylhexanoic acid, 4-methylhexanoic acid, 5-methylhexanoic acid, 2-hexenoic acid, 3- hexenoic acid, 4-hexenoic acid, 5-hexenoic acid, 5-chlorovaleric acid, 5-aminovaleric acid, 5-cyanovaleric acid, 5-(methylsulfanyl)valeric acid, 5-hydroxyvaleric acid, 5-phenylvaleric acid, 2,3-dimethylhexanoic acid, d3-hexanoic acid, 4-pentynoic acid, trans-2-pentenoic acid, 5-hexynoic acid, trans-2-hexenoic acid, 6-heptynoic acid, trans-2-octenoic acid, trans-2- nonenoic acid, 4-phenylbutyric acid, 6-phenylhexanoic acid, 7-phenyly heptanoic acid, and the like In some embodiments, the carboxylic acid is 2-methylhexanoic acid In some embodiments, the carboxylic acid is 3-methylhexanoic acid. In some embodiments, the carboxylic acid is 4-methylhexanoic acid. In some embodiments, the carboxylic acid is 5- methylhexanoic acid. In some embodiments, the carboxylic acid is 2-hexenoic acid. In some embodiments, the carboxylic acid is 3-hexenoic acid. In some embodiments, the carboxylic acid is 4-hexenoic acid. In some embodiments, the carboxylic acid is 5-hexenoic acid. In some embodiments, the carboxylic acid is 5-chlorovaleric acid. In some embodiments, the carboxylic acid is 5-aminovaleric acid. In some embodiments, the carboxylic acid is 5-cyanovaleric acid. In some embodiments, the carboxylic acid is 5- (methylsulfanyl)valeric acid. In some embodiments, the carboxylic acid is 5-hydroxyvaleric acid. In some embodiments, the carboxylic acid is 5-phenylvaleric acid. In some embodiments, the carboxylic acid is 2,3-dimethylhexanoic acid. In some embodiments, the carboxylic acid is dx-hexanoic acid. In some embodiments, the carboxylic acid is 4- pentynoic acid. In some embodiments, the carboxylic acid is trans-2-pentenoic acid. In some embodiments, the carboxylic acid is 5-hexynoic acid. In some embodiments, the carboxylic acid is trans-2-hexenoic acid. In some embodiments, the carboxylic acid is 6-heptynoic acid. In some embodiments, the carboxylic acid is trans-2-octenoic acid. In some embodiments, the carboxylic acid is trans-2-nonenoic acid. In some embodiments, the carboxylic acid is 4- phenylbutyric acid. In some embodiments, the carboxylic acid is 6-phenylhexanoic acid. In some embodiments, the carboxylic acid is 7-phenylheptanoic acid.

[001083] In some embodiments wherein the modified host cell of the present disclosure is cultured in a culture medium comprising a carboxylic acid, the carboxylic acid is an unsubstituted or substituted C3-C 18 carboxylic acid. In certain such embodiments, the unsubstituted or substituted C3-C 18 carboxylic acid is an unsubstituted or substituted hexanoic acid. In some embodiments, the cannabinoid or cannabinoid derivative is produced in an amount of more than 100 mg/L culture medium. In some embodiments, the

cannabinoid or cannabinoid derivative is produced in an amount of more than 50 mg/L culture medium.

[001084] In some embodiments wherein the modified host cell of the present disclosure is cultured in a culture medium comprising a carboxylic acid, the carboxylic acid is butyric acid, valeric acid, hexanoic acid, octanoic acid, 2-methylhexanoic acid, 3-methylhexanoic acid, 4-methylhexanoic acid, 5-methylhexanoic acid, 2-hexenoic acid, 3-hexenoic acid, 4- hexenoic acid, 5-hexenoic acid, heptanoic acid, 5-chlorovaleric acid, 5- (methylsulfanyl)valeric acid, 4-pentynoic acid, trans-2-pentenoic acid, 5-hexynoic acid, trans-2-hexenoic acid, 6-heptynoic acid, trans-2-octenoic acid, nonanoic acid, trans-2- nonenoic acid, decanoic acid, undecanoic acid, dodecanoic acid, myristic acid, 4- phenylbutyric acid, 5-phenylvaleric acid, 6-phenylhexanoic acid, 7-phenylheptanoic acid, isobutyric acid, fumaric acid, itaconic acid, malic acid, succinic acid, maleic acid, malonic acid, glutaric acid, glucaric acid, oxalic acid, adipic acid, pimelic acid, suberic acid, azelaic acid, sebacic acid, dodecandioic acid, glutaconic acid, ortho-phthalic acid, isophthalic acid, terephthalic acid, citric acid, isocitric acid, aconitic acid, tricarballylic acid, trimesic acid, 5- aminovaleric acid, 5-cyanovaleric acid, 5-hydroxyvaleric acid, or 2,3 -dimethylhexanoic acid. In some embodiments, the cannabinoid or cannabinoid derivative is produced in an amount of more than 100 mg/L culture medium. In some embodiments, the cannabinoid or cannabinoid derivative is produced in an amount of more than 50 mg/L culture medium.

[001085] In some embodiments wherein the modified host cell of the present disclosure is cultured in a culture medium comprising a carboxylic acid, the carboxylic acid is butyric acid, valeric acid, hexanoic acid, octanoic acid, 2-methylhexanoic acid, 3-methylhexanoic acid, 4-methylhexanoic acid, 5-methylhexanoic acid, 2-hexenoic acid, 3-hexenoic acid, 4- hexenoic acid, 5-hexenoic acid, heptanoic acid, 5-chlorovaleric acid, 5- (methylsulfanyl)valeric acid, 4-pentynoic acid, trans-2-pentenoic acid, 5-hexynoic acid, trans-2 -hexenoic acid, 6-heptynoic acid, trans-2-octenoic acid, nonanoic acid, trans-2 - nonenoic acid, decanoic acid, undecanoic acid, dodecanoic acid, myristic acid, 4- phenylbutyric acid, 5-phenylvaleric acid, 6-phenylhexanoic acid, 7-phenylheptanoic acid, isobutyric acid, fumaric acid, succinic acid, maleic acid, malonic acid, glutaric acid, oxalic acid, adipic acid, pimelic acid, suberic acid, azelaic acid, sebacic acid, dodecandioic acid, ortho-phthalic acid, isophthalic acid, terephthalic acid, trimesic acid, 5-aminovaleric acid, 5- cyanovaleric acid, 5-hydroxyvaleric acid, or 2,3-dimethylhexanoic acid. In some embodiments, the cannabinoid or cannabinoid derivative is produced in an amount of more than 100 mg/L culture medium. In some embodiments, the cannabinoid or cannabinoid derivative is produced in an amount of more than 50 mg/L culture medium.

[001086] In some embodiments wherein the modified host cell of the present disclosure is cultured in a culture medium comprising a carboxylic acid, the carboxylic acid is 2- methylhexanoic acid, 3-methylhexanoic acid, 4-methylhexanoic acid, 5-methylhexanoic acid, 2-hexenoic acid, 3-hexenoic acid, 4-hexenoic acid, heptanoic acid, 5-chlorovaleric acid, 5-(methylsulfanyl)valeric acid, 4-pentynoic acid, trans-2-pentenoic acid, 5-hexynoic acid, trans-2-hexenoic acid, 6-heptynoic acid, trans-2-octenoic acid, nonanoic acid, trans-2- nonenoic acid, decanoic acid, undecanoic acid, dodecanoic acid, myristic acid, 4- phenylbutyric acid, 5-phenylvaleric acid, 6-phenylhexanoic acid, 7-phenylheptanoic acid, isobutyric acid, fumaric acid, itaconic acid, malic acid, maleic acid, glucaric acid, suberic acid, azelaic acid, sebacic acid, dodecandioic acid, glutaconic acid, ortho-phthalic acid, isophthalic acid, terephthalic acid, citric acid, isocitric acid, aconitic acid, tricarballylic acid, trimesic acid, 5-aminovaleric acid, 5-cyanovaleric acid, 5-hydroxyvaleric acid, or 2,3- dimethylhexanoic acid. In some embodiments, the cannabinoid or cannabinoid derivative is produced in an amount of more than 100 mg/L culture medium. In some embodiments, the cannabinoid or cannabinoid derivative is produced in an amount of more than 50 mg/L culture medium.

[001087] In some embodiments wherein the modified host cell of the present disclosure is cultured in a culture medium comprising a carboxylic acid, the carboxylic acid is 2- methylhexanoic acid, 3-methylhexanoic acid, 4-methylhexanoic acid, 5-methylhexanoic acid, 2-hexenoic acid, 3-hexenoic acid, 4-hexenoic acid, heptanoic acid, 5-chlorovaleric acid, 5-(methylsulfanyl)valeric acid, 4-pentynoic acid, trans-2-pentenoic acid, 5-hexynoic acid, trans-2-hexenoic acid, 6-heptynoic acid, trans-2-octenoic acid, nonanoic acid, trans-2- nonenoic acid, decanoic acid, undecanoic acid, dodecanoic acid, myristic acid, 4- phenylbutyric acid, 5-phenylvaleric acid, 6-phenylhexanoic acid, 7-phenylheptanoic acid, isobutyric acid, fumaric acid, maleic acid, suberic acid, azelaic acid, sebacic acid, dodecandioic acid, ortho-phthalic acid, isophthalic acid, terephthalic acid, trimesic acid, 5- aminovaleric acid, 5-cyanovaleric acid, 5-hydroxyvaleric acid, or 2,3 -dimethylhexanoic acid. In some embodiments, the cannabinoid or cannabinoid derivative is produced in an amount of more than 100 mg/L culture medium. In some embodiments, the cannabinoid or cannabinoid derivative is produced in an amount of more than 50 mg/L culture medium.

[001088] In some embodiments wherein the modified host cell of the present disclosure is cultured in a culture medium comprising a carboxylic acid, the carboxylic acid is 4- pentynoic acid, trans-2-pentenoic acid, 5-hexynoic acid, trans-2-hexenoic acid, 6-heptynoic acid, trans-2-octenoic acid, nonanoic acid, trans-2-nonenoic acid, decanoic acid, undecanoic acid, dodecanoic acid, 4-phenylbutyric acid, 5-phenylvaleric acid, 6-phenylhexanoic acid, or 7-phenylheptanoic acid. In some embodiments, the cannabinoid or cannabinoid derivative is produced in an amount of more than 100 mg/L culture medium. In some embodiments, the cannabinoid or cannabinoid derivative is produced in an amount of more than 50 mg/L culture medium.

[001089] In some embodiments wherein the modified host cell of the present disclosure is cultured in a culture medium comprising a carboxylic acid, the carboxylic acid is 2- methylhexanoic acid, 4-methylhexanoic acid, 5-methylhexanoic acid, 2-hexenoic acid, 3- hexenoic acid, 4-hexenoic acid, heptanoic acid, 5-chlorovaleric acid, or 5- (methylsulfanyl)valeric acid. In some embodiments, the cannabinoid or cannabinoid derivative is produced in an amount of more than 100 mg/L culture medium. In some embodiments, the cannabinoid or cannabinoid derivative is produced in an amount of more than 50 mg/L culture medium.

[001090] In some embodiments wherein the modified host cell of the present disclosure is cultured in a culture medium comprising a carboxylic acid, the carboxylic acid is a carboxylic acid of Formula (II):

wherein R is a Ci-C ixalkyl group substituted with R^a or R is a C2-Cisalkenyl group substituted with R^a, wherein R^a is -S-Ci-C6alkyl or -S-C2-C6alkenyl. In some embodiments, the cannabinoid or cannabinoid derivative is produced in an amount of more than 100 mg/L culture medium. In some embodiments, the cannabinoid or cannabinoid derivative is produced in an amount of more than 50 mg/L culture medium.

[001091] The disclosure also provides methods of producing a cannabinoid or a cannabinoid derivative, such as those described herein, the method comprising: culturing a modified host cell of the disclosure in a culture medium comprising olivetolic acid or an olivetolic acid derivative. In certain such embodiments, the method comprises recovering the produced cannabinoid or cannabinoid derivative. In certain such embodiments, the produced cannabinoid or cannabinoid derivative is then purified as disclosed herein.

[001092] Olivetolic acid derivatives used herein may be substituted with or comprise one or more reactive and/or functional groups as disclosed herein. In some embodiments, an olivetolic acid derivative may lack one or more chemical moieties found in olivetolic acid. In some embodiments when the culture medium comprises an olivetolic acid derivative, the olivetolic acid derivative is orsellinic acid. In some embodiments when the culture medium comprises an olivetolic acid derivative, the olivetolic acid derivative is divarinic acid. In some embodiments, the cannabinoid or cannabinoid derivative is produced in an amount of more than 100 mg/L culture medium. In some embodiments, the cannabinoid or cannabinoid derivative is produced in an amount of more than 50 mg/L culture medium.

Exemylarv Cell Culture Conditions

[001093] Suitable media for culturing modified host cells of the disclosure may include standard culture media (e.g., Luria-Bertani broth, optionally supplemented with one or more additional agents, such as an inducer (e.g., where nucleic acids disclosed herein are under the control of an inducible promoter, etc ); standard yeast culture media; and the like). In some embodiments, the culture medium can be supplemented with one or more fermentable sugars (e.g., a hexose sugar or a pentose sugar, e.g., glucose, xylose, galactose, and the like). In some embodiments, the culture medium can be supplemented with unsubstituted or substituted hexanoic acid, carboxylic acids other than unsubstituted or substituted hexanoic acid, olivetolic acid, or olivetolic acid derivatives. In some embodiments, the culture medium can be supplemented with pretreated cellulosic feedstock (e.g., wheat grass, wheat straw, barley straw, sorghum, rice grass, sugarcane straw, bagasse, switchgrass, corn stover, corn fiber, grains, or any combination thereof). In some embodiments, the culture medium can be supplemented with oleic acid. In some embodiments, the culture medium comprises a non-fermentable carbon source. In certain such embodiments, the non-fermentable carbon source comprises ethanol. In some embodiments, the suitable media comprises an inducer. In certain such embodiments, the inducer comprises galactose. In some embodiments, the inducer comprises KH2PO4, galactose, glucose, sucrose, maltose, an amino acid (e g., methionine, lysine), CuSCri, a change in temperature (e.g., 30 °C to 37 °C), a change in pH (e.g., pH 6 to pH 4), a change in oxygen level (e.g., 20% to 1% dissolved oxygen levels), addition of hydrogen peroxide or superoxide-generating drug menadione, tunicamycin, expression of proteins prone to misfolding (e.g., cannabinoid synthases), estradiol, or doxycycline. Additional induction systems are detailed herein.

[001094] The carbon source in the suitable media can vary significantly, from simple sugars like glucose to more complex hydrolysates of other biomass, such as yeast extract. The addition of salts generally provide essential elements such as magnesium, nitrogen, phosphorus, and sulfur to allow the cells to synthesize polypeptides and nucleic acids. The suitable media can also be supplemented with selective agents, such as antibiotics, to select for the maintenance of certain plasmids and the like. For example, if a microorganism is resistant to a certain antibiotic, such as ampicillin or tetracycline, then that antibiotic can be added to the medium in order to prevent cells lacking the resistance from growing. The suitable media can be supplemented with other compounds as necessary to select for desired physiological or biochemical characteristics, such as particular amino acids and the like.

[001095] In some embodiments, modified host cells disclosed herein are grown in minimal medium or minimal media. As used herein, the terms“minimal medium” or “minimal media” may refer to media comprising a defined composition of nutrients, generally chosen for minimal cost, while still allowing for robust growth and production. As used herein, the terms“minimal medium” or“minimal media” may refer to media containing: (1) one or more carbon sources for cellular (e.g., bacterial or yeast) growth; (2) various salts, which can vary among cellular (e.g., bacterial or yeast) species and growing conditions; (3) vitamins and trace elements; and (4) water. Generally, but not always, minimal media lacks one or more amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acids). Minimal media may also comprise growth factors, inducers, and repressors.

[001096] In some embodiments, modified host cells disclosed herein are grown in defined medium or defined media (also referred to as defined minimal media or defined minimal medium). As used herein, the terms“defined medium” or“defined media” may refer to growth medium in which the exact chemical composition is known. This composition may be in excess of what is minimally required for growth.

[001097] In some embodiments, the minimal media or minimal medium or defined medium or defined media comprises galactose, glucose, ammonium sulfate, potassium phosphate, magnesium sulfate, succinate, D-biotin, Ca-D-pantothenate, nicotinic acid, myo inositol, thiamine hydrochloride, pyridoxal hydrochloride, p-aminobenzoic acid, EDTA, zinc sulfate heptahydrate, manganese chloride dihydrate, cobalt chloride hexahydrate, copper sulfate pentahydrate, sodium molybdate dihydrate, calcium chloride dihydrate, and iron sulfate heptahydrate. In certain such embodiments, the minimal media or minimal medium or defined medium or defined media comprises 40 g/L galactose, 1 g/L glucose, 15 g/L ammonium sulfate, 8 g/L potassium phosphate, 6.15 g/L magnesium sulfate, 5.9 g/L succinate, 60 pg/L D-biotin, 120 pg/L Ca-D-pantothenate, 120 pg/L nicotinic acid, 300 pg/L myo-inositol, 60 pg/L thiamine hydrochloride, 60 pg/L pyridoxal hydrochloride, 2.5 pg/L p- aminobenzoic acid, 150 mg/L EDTA, 57.5 mg/L zinc sulfate heptahydrate, 2.6 mg/L manganese chloride dihydrate, 4.7 mg/L cobalt chloride hexahydrate, 5 mg/L copper sulfate pentahydrate, 4.8 mg/L sodium molybdate dihydrate, 29 mg/L calcium chloride dihydrate, and 28 mg/L iron sulfate heptahydrate. In some embodiments, the minimal medium or minimal media or defined medium or defined media comprises a carboxylic acid (e.g., 1 mM olivetolic acid, 1 mM olivetolic acid derivative, 2 mM unsubstituted or substituted hexanoic acid, or 2 mM of a carboxylic acid other than unsubstituted or substituted hexanoic acid). In some embodiments, minimal media or minimal medium or defined medium or defined media affords higher biomass formation in a fermentation tank compared to rich medium or rich media. Minimal media or minimal medium or defined medium or defined media may permit regulation of the components of the media or medium such that everything is available in the desired proportions during fermentation. By comparison, rich media or rich medium may permit accumulation of unconsumed components during fermentation, slowing or inhibiting cell growth as these components accumulate over the timecourse of the fermentation. Minimal media or minimal medium or defined medium or defined media affords more homogeneity and reproducibility between media batches, enhanced stability over time, eased technology transfer of media formulations, and enhanced product quality and purity compared to rich medium or rich media.

[001098] In some embodiments, modified host cells disclosed herein are grown in rich medium or rich media. The components of rich media or rich medium are often undefined, as the yeast extract peptone dextrose (YPD) is largely composed of undefined extracts that vary batch to batch. This variation can result in differences in bioprocess performance during fermentation. In some embodiments, the rich medium or rich media comprises yeast extract peptone dextrose (YPD) media comprising water, yeast extract, Bacto peptone, and dextrose (glucose). In certain such embodiments, the rich medium or rich media comprises yeast extract peptone dextrose (YPD) media comprising water, 10 g/L yeast extract, 20 g/L Bacto peptone, and 20 g/L dextrose (glucose). In some embodiments, the rich medium or rich media comprises YP + galactose and glucose. In some embodiments, the rich medium or rich media comprises YP + 20 g/L galactose or YP + 40 g/L galactose and 1 g/L glucose. In some embodiments, the rich medium or rich media comprises a carboxylic acid (e.g., 1 mM olivetolic acid, 1 mM olivetolic acid derivative, 2 mM unsubstituted or substituted hexanoic acid, or 2 mM of a carboxylic acid other than unsubstituted or substituted hexanoic acid). In some embodiments, rich medium or rich media affords greater cell density in fermentation compared to minimal media or minimal medium or defined media or defined medium. In some embodiments, the modified host cell can produce the cannabinoid or cannabinoid derivative when cultured in a rich medium.

[001099] Surprisingly, it was found that some modified host cells of the disclosure can produce a cannabinoid or cannabinoid derivative when cultured in a defined medium or minimal medium. It is undesirable to scale up a fermentation process that uses rich or undefined media, e.g., YP based media, as it is expensive and cannot generate as much biomass in tanks as defined or minimal media during fermentation. Additionally, rich media causes more foaming during fermentation in tanks compared to defined or minimal media, requiring the addition of more anti-foam reagents to the culture and limiting the degree to which the tank is filled. Moreover, there is inherent variability between batches or lots of rich media, which may result in issues in the consistency of the fermentation process. Also, with minimal media or minimal medium or defined medium or defined media, the feed solutions are typically a lOOx or lOOOx stock, but for rich medium or rich media, the standard feeds are usually 2x to 4x stock. Therefore, minimal media or minimal medium or defined medium or defined media is advantageous as it requires feeding significantly lower volumes of liquid into the fermentation tank, keeping cell density higher and avoiding filling the fermentation tank too quickly. With rich medium, the feed fills the fermentation tank quickly, requiring draining and refilling of the tank, constantly washing out and diluting the cells therein. In some embodiments, the modified host cell can produce a cannabinoid or cannabinoid derivative when cultured in a defined medium or minimal medium.

[001100] In some embodiments, the modified host cell that can produce a cannabinoid or cannabinoid derivative when cultured in a defined medium comprises:

a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMG-CoA synthase (HMGS) polypeptide;

i) one or more heterologous nucleic acid comprising a nucleotide sequence encoding a pyruvate decarboxylase (PDC) polypeptide;

In certain such embodiments, the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide, one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide, and/or one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acyl-activating enzyme (AAE) polypeptide. In some embodiments, the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. In some embodiments, the nucleotide sequence are codon-optimized.

[001101] In some embodiments, the modified host cell that can produce a cannabinoid or cannabinoid derivative when cultured in a defined medium comprises:

k) two or moreheterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate :olivetolic acid geranyltransferase (GOT) polypeptide; and l) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise:

4) one or more heterologous nucleic acid comprising a nucleotide sequence encoding a protein disulfide isomerase polypeptide, wherein the protein disulfide isomerase polypeptide is a PDI1 polypeptide.

In certain such embodiments, the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide, one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide, and/or one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acyl-activating enzyme (AAE) polypeptide. In some embodiments, the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide. In some embodiments, the nucleotide sequences are codon-optimized.

[001102] In some embodiments, the modified host cell that can produce a cannabinoid or cannabinoid derivative when cultured in a defined medium comprises:

[001103] In some embodiments, the modified host cell that can produce a cannabinoid or cannabinoid derivative when cultured in a defined medium comprises:

k) two heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate:olivetolic acid geranyltransferase (GOT) polypeptide; and 1) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise:

[001104] Materials and methods suitable for the maintenance and growth of the recombinant cells of the disclosure are described herein, e.g., in the Examples section. Other materials and methods suitable for the maintenance and growth of cell (e.g., bacterial or yeast) cultures are well known in the art. Exemplary techniques can be found in International Publication No. W02009/076676, U.S. Patent Application No. 12/335,071 (U.S. Publ. No. 2009/0203102), WO 2010/003007, US Publ. No. 2010/0048964, WO 2009/132220, US Publ. No. 2010/0003716, Manual of Methods for General Bacteriology Gerhardt et al, eds), American Society for Microbiology, Washington, D.C. (1994) or Brock in Biotechnology: A Textbook of Industrial Microbiology, Second Edition (1989) Sinauer Associates, Inc., Sunderland, MA. [001105] Standard cell culture conditions can be used to culture the modified host cells disclosed herein (see, for example, WO 2004/033646 and references cited therein). In some embodiments, cells are grown and maintained at an appropriate temperature, gas mixture, and pH (such as at about 20 °C to about 37 °C, at about 0.04% to about 84% CO2, at about 0% to about 100% dissolved oxygen, and at a pH between about 2 to about 9). In some embodiments, modified host cells disclosed herein are grown at about 34 °C in a suitable cell culture medium. In some embodiments, modified host cells disclosed herein are grown at about 20 °C to about 37 °C in a suitable cell culture medium. While the growth optimum for S. cerevisiae is about 30 °C, culturing cells at a higher temperature, e.g., 34 °C may be advantageous by reducing the costs to cool industrial fermentation tanks. In some embodiments, modified host cells disclosed herein are grown at about 20 °C, about 21 °C, about 22 °C, about 23 °C, about 24 °C, about 25 °C, about 26 °C, about 27 °C, about 28 °C, about 29 °C, about 30 °C, about 31 °C, about 32 °C, about 33 °C, about 34 °C, about 35 °C, about 36 °C, or about 37 °C in a suitable cell culture medium. In some embodiments, the pH ranges for fermentation are between about pH 3.0 to about pH 9.0 (such as about pH 3.0, about pH 3.5, about pH 4.0, about pH 4.5, about pH 5.0, about pH 5.5, about pH 6.0, about pH 6.5, about pH 7.0, about pH 7.5, about pH 8.0, about pH 8.5, about pH 6.0 to about pH 8.0 or about pH 6.5 to about pH 7.0). In some embodiments, the pH ranges for fermentation are between about pH 4.5 to about pH 5.5. In some embodiments, the pH ranges for fermentation are between about pH 4.0 to about pH 6.0. In some embodiments, the pH ranges for fermentation are between about pH 3.0 to about pH 6.0. In some embodiments, the pH ranges for fermentation are between about pH 3.0 to about pH 5.5. In some embodiments, the pH ranges for fermentation are between about pH 3.0 to about pH 5.0. In some embodiments, the dissolved oxygen is between about 0% to about 10%, about 0% to about 20%, about 0% to about 30%, about 0% to about 40%, about 0% to about 50%, about 0% to about 60%, about 0% to about 70%, about 0% to about 80%, about 0% to about 90%, about 5% to about 10%, about 5% to about 20%, about 5% to about 30%, about 5% to about 40%, about 5% to about 50%, about 5% to about 60%, about 5% to about 70%, about 5% to about 80%, about 5% to about 90%, about 10% to about 20%, about 10% to about 30%, about 10% to about 40% or about 10% to about 50%. In some embodiments, the CO2 level is between about 0.04% to about 0.1% CO2, about 0.04% to about 1% CO2, about 0.04% to about 5% CO2, about 0.04% to about 10% CO2, about 0.04% to about 20% CO2, about 0.04% to about 30% CO2, about 0.04% to about 40% CO2, about 0.04% to about 50% CO2, about 0.04% to about 60% CO2, about 0.04% to about 70% CO2, about 0.1% to about 5% CO2, about 0.1% to about 10% CO2, about 0.1% to about 20% CO2, about 0.1% to about 30% CO2, about 0.1% to about 40% CO2, about 0.1% to about 50% CO2, about 1% to about 5% CO2, about 1% to about 10% CO2, about 1% to about 20% CO2, about 1% to about 30% CO2, about 1% to about 40% CO2, about 1% to about 50% CO2, about 5% to about 10% CO2, about 10% to about 20% CO2, about 10% to about 30% CO2, about 10% to about 40%

CO2, about 10% to about 50% CO2, about 10% to about 60% CO2, about 10% to about 70%

CO2, about 10% to about 80% CO2, about 50% to about 60% CO2, about 50% to about 70%

CO2, or about 50% to about 80% CO2. Modified host cells disclosed herein disclosed herein can be grown under aerobic, anoxic, microaerobic, or anaerobic conditions based on the requirements of the cells.

[001106] Standard culture conditions and modes of fermentation, such as batch, fed- batch, or continuous fermentation that can be used are described in International Publication No. WO 2009/076676, U.S. Patent Application No. 12/335,071 (U.S. Publ. No.

2009/0203102), WO 2010/003007, US Publ. No. 2010/0048964, WO 2009/132220, US Publ. No. 2010/0003716, the contents of each of which are incorporated by reference herein in their entireties. Batch and Fed-Batch fermentations are common and well known in the art and examples can be found in Brock, Biotechnology: A Textbook of Industrial

Microbiology, Second Edition (1989) Sinauer Associates, Inc.

Production and Recovery of Produced Cannabinoids or Cannabinoid Derivatives

[001107] The present disclosure provides for production of a cannabinoid or a cannabinoid derivative. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative, such as those disclosed herein, by modified host cells of the disclosure in an amount of from about 1 mg/L culture medium to about 1 g/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 1 mg/L culture medium to about 500 mg/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 1 mg/L culture medium to about 100 mg/L culture medium. For example, in some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 1 mg/L culture medium to about 5 mg/L culture medium, from about 5 mg/L culture medium to about 10 mg/L culture medium, from about 10 mg/L culture medium to about 25 mg/L culture medium, from about 25 mg/L culture medium to about 50 mg/L culture medium, from about 50 mg/L culture medium to about 75 mg/L culture medium, or from about 75 mg/L culture medium to about 100 mg/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 100 mg/L culture medium to about 150 mg/L culture medium, from about 150 mg/L culture medium to about 200 mg/L culture medium, from about 200 mg/L culture medium to about 250 mg/L culture medium, from about 250 mg/L culture medium to about 500 mg/L culture medium, from about 500 mg/L culture medium to about 750 mg/L culture medium, or from about 750 mg/L culture medium to about 1 g/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about from about 50 mg/L culture medium to about 100 mg/L culture medium, 50 mg/L culture medium to about 150 mg/L culture medium, from about 50 mg/L culture medium to about 200 mg/L culture medium, from about 50 mg/L culture medium to about 250 mg/L culture medium, from about 50 mg/L culture medium to about 500 mg/L culture medium, or from about 50 mg/L culture medium to about 750 mg/L culture medium.

[001108] In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative, such as those disclosed herein, in an amount of from about 50 mg/L culture medium to about 100 g/L culture medium, or more than 100 g/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative, such as those disclosed herein, in an amount of from about 50 mg/L culture medium to about 100 mg/L culture medium, or more than 100 mg/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative, such as those disclosed herein, in an amount of more than 50 mg/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative, such as those disclosed herein, in an amount of more than 100 mg/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 100 mg/L culture medium to about 500 mg/L culture medium, or more than 500 mg/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 500 mg/L culture medium to about 1 g/L culture medium, or more than 1 g/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 1 g/L culture medium to about 10 g/L culture medium, or more than 10 g/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 10 g/L culture medium to about 100 g/L culture medium, or more than 100 g/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 1 g/L culture medium to about 20 g/L culture medium, or more than 20 g/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 1 g/L culture medium to about 30 g/L culture medium, or more than 30 g/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 1 g/L culture medium to about 40 g/L culture medium, or more than 40 g/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 1 g/L culture medium to about 50 g/L culture medium, or more than 50 g/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 1 g/L culture medium to about 60 g/L culture medium, or more than 60 g/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 1 g/L culture medium to about 70 g/L culture medium, or more than 70 g/L culture medium.

In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 1 g/L culture medium to about 80 g/L culture medium, or more than 80 g/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 1 g/L culture medium to about 90 g/L culture medium, or more than 90 g/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 10 g/L culture medium to about 20 g/L culture medium, or more than 20 g/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 10 g/L culture medium to about 30 g/L culture medium, or more than 30 g/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 10 g/L culture medium to about 40 g/L culture medium, or more than 40 g/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 10 g/L culture medium to about 50 g/L culture medium, or more than 50 g/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 10 g/L culture medium to about 60 g/L culture medium, or more than 60 g/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 10 g/L culture medium to about 70 g/L culture medium, or more than 70 g/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 10 g/L culture medium to about 80 g/L culture medium, or more than 80 g/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 10 g/L culture medium to about 90 g/L culture medium, or more than 90 g/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 50 g/L culture medium to about 100 g/L culture medium, or more than 100 g/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 50 g/L culture medium to about 60 g/L culture medium, or more than 60 g/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 50 g/L culture medium to about 70 g/L culture medium, or more than 70 g/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 50 g/L culture medium to about 80 g/L culture medium, or more than 80 g/L culture medium. In some

embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 50 g/L culture medium to about 90 g/L culture medium, or more than 90 g/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 20 g/L culture medium to about 100 g/L culture medium, or more than 100 g/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 20 g/L culture medium to about 30 g/L culture medium, or more than 30 g/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 20 g/L culture medium to about 40 g/L culture medium, or more than 40 g/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 20 g/L culture medium to about 50 g/L culture medium, or more than 50 g/L culture medium. In some

embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 20 g/L culture medium to about 60 g/L culture medium, or more than 60 g/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 20 g/L culture medium to about 70 g/L culture medium, or more than 70 g/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 20 g/L culture medium to about 80 g/L culture medium, or more than 80 g/L culture medium. In some embodiments, a method of the present disclosure provides for production of a cannabinoid or a cannabinoid derivative in an amount of from about 20 g/L culture medium to about 90 g/L culture medium, or more than 90 g/L culture medium.

[001109] In some embodiments, the modified host cell disclosed herein is cultured in a liquid medium comprising a carboxylic acid, olivetolic acid, or an olivetolic acid derivative.

[001110] In some embodiments, a method of producing a cannabinoid or a cannabinoid derivative, such as those disclosed herein, may involve culturing a modified yeast cell of the present disclosure under conditions that favor production of a cannabinoid or a cannabinoid derivative; wherein the cannabinoid or the cannabinoid derivative is produced by the modified yeast cell and is present in the culture medium (e g , a liquid culture medium) in which the modified yeast cell is cultured. In some embodiments, the culture medium in which the modified yeast cell is cultured comprises a cannabinoid or a cannabinoid derivative in an amount of from 1 ng/L to 1 g/L (e.g., from 1 ng/L to 50 ng/L, from 50 ng/L to 100 ng/L, from 100 ng/L to 500 ng/L, from 500 ng/L to 1 pg/L, from 1 pg/L to 50 pg/L, from 50 pg/L to 100 pg/L, from 100 pg/L to 500 pg/L, from 500 pg/L to 1 mg/L, from 1 mg/L to 50 mg/L, from 50 mg/L to 100 mg/L, from 100 mg/L to 500 mg/L, or from 500 mg/L to 1 g/L). In certain such embodiments, the modified yeast cell is a modified S.

cerevisiae. In some embodiments, the culture medium in which the modified yeast cell is cultured comprises a cannabinoid or a cannabinoid derivative in an amount from 50 mg/L to 100 mg/L. In certain such embodiments, the modified yeast cell is a modified S. cerevisiae.

In some embodiments, the culture medium in which the modified yeast cell is cultured comprises a cannabinoid or a cannabinoid derivative in an amount from 100 mg/L to 500 mg/L. In certain such embodiments, the modified yeast cell is a modified S. cerevisiae. In some embodiments, the culture medium in which the modified yeast cell is cultured comprises a cannabinoid or a cannabinoid derivative in an amount from 500 mg/L to 1 g/L. In certain such embodiments, the modified yeast cell is a modified S. cerevisiae. In some embodiments, the culture medium in which the modified yeast cell is cultured comprises a cannabinoid or a cannabinoid derivative in an amount more than 1 g/L. In certain such embodiments, the modified yeast cell is a modified S. cerevisiae.

[001111] In some embodiments, a method of producing a cannabinoid or a cannabinoid derivative, such as those disclosed herein, may involve culturing a modified yeast cell of the present disclosure under conditions that favor fermentation of a sugar, and under conditions that favor production of a cannabinoid or a cannabinoid derivative; wherein the cannabinoid or the cannabinoid derivative is produced by the modified yeast cell and is present in alcohol produced by the modified yeast cell. The present disclosure provides an alcoholic beverage produced by the modified yeast cell, where the alcoholic beverage comprises the cannabinoid or cannabinoid derivative produced by the modified yeast cell. Alcoholic beverages may include beer, wine, and distilled alcoholic beverages. In some embodiments, an alcoholic beverage of the present disclosure comprises a cannabinoid or a cannabinoid derivative in an amount of from 1 ng/L to 1 g/L (e.g., from 1 ng/L to 50 ng/L, from 50 ng/L to 100 ng/L, from 100 ng/L to 500 ng/L, from 500 ng/L to 1 pg/L, from 1 pg/L to 50 pg/L, from 50 pg/L to 100 pg/L, from 100 pg/L to 500 pg/L, from 500 pg/L to 1 mg/L, from 1 mg/L to 50 mg/L, from 50 mg/L to 100 mg/L, from 100 mg/L to 500 mg/L, or from 500 mg/L to 1 g/L). In some embodiments, an alcoholic beverage of the present disclosure comprises a cannabinoid or a cannabinoid derivative in an amount more than 1 g/L.

[001112] The present disclosure provides a beverage produced by the modified yeast cell, where the beverage comprises the cannabinoid or cannabinoid derivative, such as those disclosed herein, produced by the modified yeast cell. In some embodiments, a beverage of the present disclosure comprises a cannabinoid or a cannabinoid derivative in an amount of from 1 ng/L to 1 g/L (e.g., from 1 ng/L to 50 ng/L, from 50 ng/L to 100 ng/L, from 100 ng/L to 500 ng/L, from 500 ng/L to 1 pg/L, from 1 pg/L to 50 pg/L, from 50 pg/L to 100 pg/L, from 100 pg/L to 500 pg/L, from 500 pg/L to 1 mg/L, from 1 mg/L to 50 mg/L, from 50 mg/L to 100 mg/L, from 100 mg/L to 500 mg/L, or from 500 mg/L to 1 g/L). In some embodiments, a beverage of the present disclosure comprises a cannabinoid or a cannabinoid derivative in an amount more than 1 g/L. In some embodiments, a beverage of the present disclosure is non-alcoholic.

[001113] In some embodiments, a method of the present disclosure provides for increased production of a cannabinoid or a cannabinoid derivative, such as those disclosed herein. In certain such embodiments, culturing of the modified host cell disclosed herein in a culture medium provides for synthesis of a cannabinoid or a cannabinoid derivative in an increased amount compared to an unmodified host cell cultured under similar conditions.

The production of a cannabinoid or a cannabinoid derivative by the modified host cells disclosed herein may be increased by about 5% to about 1,000,000 folds compared to an unmodified host cell cultured under similar conditions. The production of a cannabinoid or a cannabinoid derivative by the modified host cells disclosed herein may be increased by about 10% to about 1,000,000 folds (e.g., about 50% to about 1,000,000 folds, about 1 to about 500,000 folds, about 1 to about 50,000 folds, about 1 to about 5,000 folds, about 1 to about 1,000 folds, about 1 to about 500 folds, about 1 to about 100 folds, about 1 to about 50 folds, about 5 to about 100,000 folds, about 5 to about 10,000 folds, about 5 to about 1,000 folds, about 5 to about 500 folds, about 5 to about 100 folds, about 10 to about 50,000 folds, about 50 to about 10,000 folds, about 100 to about 5,000 folds, about 200 to about 1,000 folds, about 50 to about 500 folds, or about 50 to about 200 folds) compared to the production of a cannabinoid or a cannabinoid derivative by unmodified host cells cultured under similar conditions. The production of a cannabinoid or a cannabinoid derivative by modified host cells disclosed herein may also be increased by at least about any of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 1 fold, 2 folds, 5 folds, 10 folds, 20 folds, 50 folds, 100 folds, 200 folds, 500 folds, 1000 folds, 2000 folds, 5000 folds, 10,000 folds, 20,000 folds, 50,000 folds, 100,000 folds, 200,000 folds, 500,000 folds, or 1,000,000 folds or more compared to the production of a cannabinoid or a cannabinoid derivative by unmodified host cells cultured under similar conditions.

[001114] In some embodiments, the production of a cannabinoid or a cannabinoid derivative, such as those disclosed herein, by modified host cells of the disclosure may also be increased by at least about any of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% compared to the production of a cannabinoid or a cannabinoid derivative by unmodified host cells cultured under similar conditions. In some embodiments, the production of a cannabinoid or a cannabinoid derivative by modified host cells disclosed herein may also be increased by at least about any of 1-20%, 2-20%, 5-20%, 10-20%, 15- 20%, 1-15%, 1-10%, 2-15%, 2-10%, 5-15%, 10-15%, 1-50%, 10-50%, 20-50%, 30-50%, 40-50%, 50-100%, 50-60%, 50-70%, 50-80%, 50-90%, or 50-100% compared to the production of a cannabinoid or a cannabinoid derivative by unmodified host cells cultured under similar conditions.

[001115] In some embodiments, production of a cannabinoid or a cannabinoid derivative by modified host cells of the disclosure is determined by LC-MS analysis. In certain such embodiments, each cannabinoid or cannabinoid derivative is identified by retention time, determined from an authentic standard, and multiple reaction monitoring (MRM) transition.

[001116] In some embodiments, the modified host cell of the disclosure is yeast cell.

In certain such embodiments, the modified host cell disclosed herein is cultured in a bioreactor. In some embodiments, the modified host cell is cultured in a culture medium supplemented with unsubstituted or substituted hexanoic acid, a carboxylic acid other than unsubstituted or substituted hexanoic acid, olivetolic acid, or an olivetolic acid derivative. In some embodiments, the modified yeast cell is a modified S. cerevisiae.

[001117] In some embodiments, the cannabinoid or cannabinoid derivative, such as those disclosed herein, is recovered from a cell lysate, e.g., by lysing the modified host cell disclosed herein and recovering the cannabinoid or cannabinoid derivative derivative from the lysate. In other cases, the cannabinoid or cannabinoid derivative is recovered from the culture medium in which the modified host cell disclosed herein is cultured. In other cases, the cannabinoid or cannabinoid derivative is recovered from both the cell lysate and the culture medium. In other cases, the cannabinoid or cannabinoid derivative is recovered from a modified host cell. In other cases, the cannabinoid or cannabinoid derivative is recovered from both the modified host cell and the culture medium. In other cases, the cannabinoid or cannabinoid derivative is recovered from the cell lysate, the modified host cell, and the culture medium. In some embodiments when the cannabinoid or cannabinoid derivative is recovered from a cell lysate; from a culture medium; from a modified host cell; from both the cell lysate and the culture medium; from both the modified host cell and the culture medium; or from the cell lysate, the modified host cell, and the culture medium, the recovered cannabinoid or cannabinoid derivative is in the form of a salt. In certain such embodiments, the salt is a pharmaceutically acceptable salt. In some embodiments, the salt of the recovered cannabinoid or cannabinoid derivative is then purified as disclosed herein.

[001118] In some embodiments, the recovered cannabinoid or cannabinoid derivative, such as those disclosed herein, is then purified. In some embodiments, whole-cell broth from cultures comprising modified host cells of the disclosure may be extracted with a suitable organic solvent to afford cannabinoids or cannabinoid derivatives. Suitable organic solvents include, but are not limited to, hexane, heptane, ethyl acetate, petroleum ether, and di-ethyl ether, chloroform, and ethyl acetate. In some embodiments, the suitable organic solvent comprises hexane. In some embodiments, the suitable organic solvent may be added to the whole-cell broth from fermentations comprising modified host cells of the disclosure at a 10: 1 ratio (10 parts whole-cell broth - 1 part organic solvent) and stirred for 30 minutes. In certain such embodiments, the organic fraction may be separated and extracted twice with an equal volume of acidic water (pH 2.5). The organic layer may then be separated and dried in a concentrator (rotary evaporator or thin film evaporator under reduced pressure) to obtain crude cannabinoid or cannabinoid derivative crystals. In certain such embodiments, the crude crystals may be heated or exposed to light to decarboxylate the crude cannabinoid or cannabinoid derivative. In certain such embodiments, the crude crystals may be heated to 105 °C for 15 minutes followed by 145 °C for 55 minutes to decarboxylate the crude cannabinoid or cannabinoid derivative. In certain such embodiments, the crude crystalline product may be re-dissolved and recrystallized in a suitable solvent (e.g., n-pentane) and filtered to remove any insoluble material. In certain such embodiments, the solvent may then be removed e g., by rotary evaporation, to produce pure crystalline product.

[001119] In some embodiments, the cannabinoid or cannabinoid derivative is pure, e.g., at least about 40% pure, at least about 50% pure, at least about 60% pure, at least about 70% pure, at least about 80% pure, at least about 90% pure, at least about 95% pure, at least about 98%, or more than 98% pure, where“pure” in the context of a cannabinoid or a cannabinoid derivative may refer to a cannabinoid or a cannabinoid derivative that is free from other cannabinoids or cannabinoid derivatives, macromolecules, contaminants, etc.

Methods of Preparing Cannabinoid Synthase Polypeptides

[001120] In an aspect, the present disclosure provides methods for preparing cannabinoid synthase polypeptides. In certain such embodiments, the methods may comprise culturing a modified host cell for expressing a cannabinoid synthase polypeptide of the disclosure in a culture medium. In some embodiments, the modified host cell of the disclosure is a Pichia sp. The method can comprise isolating or purifying the expressed cannabinoid synthase polypeptide, as described herein.

[001121] In some embodiments, the method for preparing cannabinoid synthase polypeptides comprises the step of isolating or purifying the polypeptides. The cannabinoid synthase polypeptides can be expressed in modified host cells, as described herein, and isolated from the modified host cells and/or culture medium using any one or more of the well known techniques used for protein purification, including, among others, lysozyme treatment, sonication, filtration, salting-out, ultra-centrifugation, and chromatography.

Chromatographic techniques for isolation of the cannabinoid synthase polypeptides may include, among others, reverse phase chromatography high performance liquid

chromatography, ion exchange chromatography, gel electrophoresis, and affinity

chromatography. In some embodiments, affinity chromatography is used.

[001122] In some embodiments, the cannabinoid synthase polypeptides expressed in the modified host cells of the disclosure can be prepared and used in various forms including but not limited to crude extracts (e g., cell-free lysates), powders (e.g., shake-flask powders), lyophilizates, frozen stocks made with glycerol or another cryoprotectant, and substantially pure preparations (e.g., DSP powders).

[001123] In some embodiments, the cannabinoid synthase polypeptides expressed in the modified host cells of the disclosure can be prepared and used in purified form.

Generally, conditions for purifying a particular polypeptide will depend, in part, on factors such as net charge, hydrophobicity, hydrophilicity, molecular weight, molecular shape, etc., and will be apparent to those having skill in the art.

Cell-Free Methods of Producing Cannabinoids or Cannabinoid Derivatives

[001124] The methods of the disclosure may involve cell-free production of cannabinoids or cannabinoid derivatives, such as those disclosed herein, using cannabinoid synthase polypeptides disclosed herein expressed or overexpressed by a modified host cell of the disclosure. In some embodiments, a cannabinoid synthase polypeptide disclosed herein expressed or overexpressed by a modified host cell of the disclosure are used in a cell-free system for the production of cannabinoids or cannabinoid derivatives. In certain such embodiments, appropriate starting materials for use in producing cannabinoids or cannabinoid derivatives may be mixed together with cannabinoid synthase polypeptides disclosed herein expressed or overexpressed by a modified host cell of the disclosure in a suitable reaction vessel to effect the reaction. The cannabinoid synthase polypeptides disclosed herein expressed or overexpressed by a modified host cell of the disclosure may be used in combination to effect a complete synthesis of a cannabinoid or cannabinoid derivative from the appropriate starting materials. In some embodiments, the cannabinoid or cannabinoid derivative is recovered from a cell-free reaction mixture comprising

cannabinoid synthase polypeptides disclosed herein. [001125] In some embodiments, the recovered cannabinoids or cannabinoid

derivatives, such as those disclosed herein, are then purified. In certain such embodiments, a cell-free reaction mixture comprising cannabinoid synthase polypeptides disclosed herein may be extracted with a suitable organic solvent to afford cannabinoids or cannabinoid derivatives. Suitable organic solvents include, but are not limited to, hexane, heptane, ethyl acetate, petroleum ether, and di-ethyl ether, chloroform, and ethyl acetate. In some embodiments, the suitable organic solvent comprises hexane. In some embodiments, the suitable organic solvent may be added to the cell-free reaction mixture comprising one or more of the polypeptides disclosed herein at a 10: 1 ratio (10 parts reaction mixture - 1 part organic solvent) and stirred for 30 minutes. In certain such embodiments, the organic fraction may be separated and extracted twice with an equal volume of acidic water (pH 2.5). The organic layer may then be separated and dried in a concentrator (rotary evaporator or thin film evaporator under reduced pressure) to obtain crude cannabinoid or cannabinoid derivative crystals. In certain such embodiments, the crude crystals may be heated or exposed to light to decarboxylate the crude cannabinoid or cannabinoid derivative. In certain such embodiments, the crude crystals may be heated to 105 °C for 15 minutes followed by 145 °C for 55 minutes to decarboxylate the crude cannabinoid or cannabinoid derivative. In certain such embodiments, the crude crystalline product may be re-dissolved and

recrystallized in a suitable solvent (e.g., n-pentane) and filtered to remove any insoluble material. In certain such embodiments, the solvent may then be removed e g., by rotary evaporation, to produce pure crystalline product.

[001126] In some embodiments, cell-free production of a cannabinoid or a cannabinoid derivative by cannabinoid synthase polypeptides disclosed herein expressed or

overexpressed by a modified host cell of the disclosure is determined by LC-MS analysis. In certain such embodiments, each cannabinoid or cannabinoid derivative is identified by retention time, determined from an authentic standard, and multiple reaction monitoring (MRM) transition.

[001127] In some embodiments when the cannabinoid or cannabinoid derivative is recovered from a cell-free reaction mixture comprising one or more polypeptides disclosed herein, the recovered cannabinoid or cannabinoid derivative is in the form of a salt. In certain such embodiments, the salt is a pharmaceutically acceptable salt. In some

embodiments, the salt of the recovered cannabinoid or cannabinoid derivative is then purified as disclosed herein. Compounds of the Disclosure

[001128] The disclosure provides novel cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof.

[001129] In some embodiments, the cannabinoids or cannabinoid derivatives of the present disclosure are compounds of Formula (I):

or pharmaceutically acceptable salts thereof, wherein,

R¹ is a Ci-Cisalkyl group substituted with R^a or R¹ is a Cb-C ixalkenyl group substituted with R^a, wherein R^a is -S-Ci-C6alkyl or -S-C2-C6alkenyl, or R¹ is selected from the group consisting of:

[001130] In some embodiments the compound of Formula (I), or a pharmaceutically acceptable salt thereof, may include where R¹ is a Ci-Cisalkyl group substituted with R^a or R¹ is a C2-Ci8alkenyl group substituted with R^a, wherein R^a is -S-Ci-C6alkyl or -S-C2- Cealkenyl, or R¹ is selected from the group consisting of:

[001131] In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, may include where R¹ is selected from the group consisting of:

[001132] In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, may include where R¹ is a C i-Cixalkyl group substituted with R^a or R¹ is a C2-Ci8alkenyl group substituted with R^a, wherein R^a is -S-Ci-C6alkyl or -S-C2- Csalkenyl.

[001133] In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, may include where R¹ is selected from the group consisting of:

[001134] In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is selected from the group consisting of:

Ĵ86

[001135] In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is selected from the group consisting of:

[001136] In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is selected from the group consisting of:

Compositions of the Disclosure

[001137] The disclosed cannabinoids and cannabinoid derivatives, or pharmaceutically acceptable salts thereof (e.g., compounds of Formula (I), or pharmaceutically acceptable salts thereof), may be used on their own but will generally be administered in the form of a composition, for example a pharmaceutical composition, in which one or more disclosed cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, is in association with an acceptable carrier, for example a pharmaceutically acceptable carrier or a carrier generally regarded as safe (GRAS). Conventional procedures for the selection and preparation of suitable pharmaceutical compositions are described in, for example, “Pharmaceuticals - The Science of Dosage Form Designs,” M. E. Aulton, Churchill Livingstone, 1988, which is hereby incorporated by reference in its entirety. The compositions described herein (e.g., pharmaceutical compositions and GRAS compositions) may be used in connection with the methods of treatment, uses, and medicaments described herein.

[001138] The term“carrier,” as used in this disclosure, may encompass carriers, excipients, and diluents and may mean a material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, involved in carrying or transporting an agent, such as one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., compounds of Formula (I), or pharmaceutically acceptable salts thereof) from one organ, or portion of the body, to another organ, or portion of the body of a subject. Carriers should be selected on the basis of compatibility and the release profile properties of the desired dosage form. Carriers may also be selected because they are GRAS. Exemplary carrier materials may include, e.g., binders, suspending agents, disintegration agents, filling agents, surfactants, solubilizers, stabilizers, lubricants, wetting agents, diluents, spray-dried dispersions, and the like. See, e.g., Hoover, John E., Remington’s Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa. 1975.

[001139] In some embodiments, the disclosure provides for a composition comprising one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., compounds of Formula (I), or pharmaceutically acceptable salts thereof) and an acceptable carrier. In some embodiments, the disclosure provides for a composition comprising a cannabinoid or cannabinoid derivative, or a pharmaceutically acceptable salt thereof, disclosed herein (e.g., compounds of Formula (I), or

pharmaceutically acceptable salts thereof) and an acceptable carrier. In some embodiments, the disclosure provides for a composition comprising two or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., compounds of Formula (I), or pharmaceutically acceptable salts thereof) and an acceptable carrier. In some embodiments, the disclosure provides for a composition comprising three or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., compounds of Formula (I), or pharmaceutically acceptable salts thereof) and an acceptable carrier. In some embodiments, the disclosure provides for a composition comprising four or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., compounds of Formula (I), or pharmaceutically acceptable salts thereof) and an acceptable carrier. In some

embodiments, the disclosure provides for a composition comprising five or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., compounds of Formula (I), or pharmaceutically acceptable salts thereof) and an acceptable carrier.

[001140] In some embodiments, the disclosure provides for a pharmaceutical composition comprising one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., compounds of Formula (I), or pharmaceutically acceptable salts thereof) and a pharmaceutically acceptable carrier. In some embodiments, the disclosure provides for a pharmaceutical composition comprising a cannabinoid or cannabinoid derivative, or a pharmaceutically acceptable salt thereof, disclosed herein (e.g., compounds of Formula (I), or pharmaceutically acceptable salts thereof) and a pharmaceutically acceptable carrier. In some embodiments, the disclosure provides for a pharmaceutical composition comprising two or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., compounds of Formula (I), or pharmaceutically acceptable salts thereof) and a

pharmaceutically acceptable carrier. In some embodiments, the disclosure provides for a pharmaceutical composition comprising three or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., compounds of Formula (I), or pharmaceutically acceptable salts thereof) and a pharmaceutically acceptable carrier. In some embodiments, the disclosure provides for a pharmaceutical composition comprising four or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., compounds of Formula (I), or pharmaceutically acceptable salts thereof) and a pharmaceutically acceptable carrier. In some embodiments, the disclosure provides for a pharmaceutical composition comprising five or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., compounds of Formula (I), or pharmaceutically acceptable salts thereof) and a pharmaceutically acceptable carrier.

[001141] In some embodiments, the disclosure provides for a composition comprising one or more compounds of Formula (I), or pharmaceutically acceptable salts thereof, and an acceptable carrier. In some embodiments, the disclosure provides for a composition comprising a compound of Formula (I), or a pharmaceutically acceptable salt thereof, and an acceptable carrier. In some embodiments, the disclosure provides for a composition comprising two or more compounds of Formula (I), or pharmaceutically acceptable salts thereof, and an acceptable carrier. In some embodiments, the disclosure provides for a composition comprising three or more compounds of Formula (I), or pharmaceutically acceptable salts thereof, and an acceptable carrier. In some embodiments, the disclosure provides for a composition comprising four or more compounds of Formula (I), or pharmaceutically acceptable salts thereof, and an acceptable carrier. In some embodiments, the disclosure provides for a composition comprising five or more compounds of Formula (I), or pharmaceutically acceptable salts thereof, and an acceptable carrier.

[001142] In some embodiments, the disclosure provides for a pharmaceutical composition comprising one or more compounds of Formula (I), or pharmaceutically acceptable salts thereof, and a pharmaceutically acceptable carrier. In some embodiments, the disclosure provides for a pharmaceutical composition comprising a compound of Formula (I), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. In some embodiments, the disclosure provides for a pharmaceutical composition comprising two or more compounds of Formula (I), or pharmaceutically acceptable salts thereof, and a pharmaceutically acceptable carrier. In some embodiments, the disclosure provides for a pharmaceutical composition comprising three or more compounds of Formula (I), or pharmaceutically acceptable salts thereof, and a pharmaceutically acceptable carrier.

In some embodiments, the disclosure provides for a pharmaceutical composition comprising four or more compounds of Formula (I), or pharmaceutically acceptable salts thereof, and a pharmaceutically acceptable carrier. In some embodiments, the disclosure provides for a pharmaceutical composition comprising five or more compounds of Formula (I), or pharmaceutically acceptable salts thereof, and a pharmaceutically acceptable carrier.

[001143] Depending on the mode of administration, the composition or pharmaceutical composition will comprise from about 0.05 to about 99 wt% (percent by weight), more particularly from about 0.05 to about 80 wt%, still more particularly from about 0.10 to about 70 wt%, and even more particularly from about 0.10 to about 50 wt% of one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof), all percentages by weight being based on total composition.

[001144] The present disclosure provides a process for the preparation of a composition of the present disclosure which comprises mixing one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) with an acceptable carrier. Compositions of the disclosure can also be prepared according to conventional mixing, granulating or coating methods.

[001145] The present disclosure provides a process for the preparation of a

pharmaceutical composition of the present disclosure which comprises mixing one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) with a pharmaceutically acceptable carrier. Pharmaceutical compositions of the disclosure can also be prepared according to conventional mixing, granulating or coating methods.

[001146] Compositions or pharmaceutical compositions of the present disclosure may comprise one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a

pharmaceutically acceptable salt thereof) formulated together with one or more acceptable carriers or pharmaceutically acceptable carriers. In some embodiments, the one or more cannabinoids or cannabinoid derivatives are present in a therapeutically effective amount. Some examples of materials that can serve as acceptable carriers or pharmaceutically acceptable carriers are sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols such a propylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer’s solution, ethyl alcohol; and phosphate buffer solutions, as well as other non toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, releasing agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the composition or

pharmaceutical composition, according to the judgment of the formulator.

[001147] Depending on the intended mode of administration, the disclosed

compositions or pharmaceutical compositions can be in solid, semi-solid or liquid dosage form, such as, for example, injectables, tablets, suppositories, pills, softgels, time-release capsules, elixirs, tinctures, oils, extracts, creams, lotions, emulsions, syrups, powders, liquids, suspensions, or the like, sometimes in unit dosages and consistent with conventional practices. These modes may include systemic or local administration such as oral, nasal, parenteral (as by intravenous (both bolus and infusion), intramuscular, or subcutaneous injection), transdermal, vaginal, buccal, rectal or topical (as by powders, ointments, or drops) administration modes. These modes may also include intracisternally, intraperitoneally, as an oral or nasal spray, or as a liquid aerosol or dry powder composition or pharmaceutical composition for inhalation.

[001148] Solid dosage forms for oral administration may include capsules, softgels, tablets, pills, powders, crystals, and granules. In such solid dosage forms, one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) may be mixed with at least one inert, acceptable or pharmaceutically acceptable carriers such as a diluent, fillers or extenders, binders, humectants, disintegrating agents, solution retarding agents, wetting agents, lubricants, an emulsifier or dispersing agent, or buffering agents.

[001149] Solid compositions or solid pharmaceutical compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.

[001150] The one or more cannabinoids or cannabinoid derivatives, or

pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) can also be in micro-encapsulated form with one or more excipients as noted herein. The solid dosage forms of tablets, dragees, softgels, capsules, pills, crystals, and granules can be prepared with coatings and shells such as enteric coatings, release controlling coatings and other coatings well known in the art. In such solid dosage forms, one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a

pharmaceutically acceptable salt thereof) may be admixed with at least one inert diluent such as sucrose, lactose or starch. Such dosage forms may also comprise, as is normal practice, additional substances other than inert diluents, e.g., tableting lubricants and other tableting aids such a magnesium stearate and microcrystalline cellulose. In the case of capsules, softgels, tablets and pills, the dosage forms may also comprise buffering agents. They may optionally contain opacifying agents and can also be of a composition that they release the one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof), only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used may include polymeric substances and waxes.

[001151] Liquid dosage forms for oral administration may include emulsions, microemulsions, solutions, suspensions, syrups, tinctures, oils, extracts, and elixirs. In addition to the one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a

pharmaceutically acceptable salt thereof), the liquid dosage forms may contain inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3 -butylene glycol, dimethylformamide, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof. Besides inert diluents, the oral compositions or oral pharmaceutical compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[001152] Injectable compositions or injectable pharmaceutical compositions, for example, sterile injectable aqueous or oleaginous suspensions comprising one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable composition or sterile injectable pharmaceutical composition may also be a sterile injectable solution, suspension or emulsion in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3- butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer’s solution, 1% lidocaine, U.S.P. and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil can be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid are used in the composition of injectables.

[001153] The injectable compositions or injectable pharmaceutical compositions can be sterilized, for example, by filtration through a bacterial-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions or sterile solid pharmaceutical compositions that can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.

[001154] In order to prolong the effect of one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof), it may desirable to slow the absorption of the one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) from subcutaneous or intramuscular injection.

This may be accomplished by the use of a liquid suspension of crystalline or amorphous material with poor water solubility. The rate of absorption of the one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) then depends upon its rate of dissolution that, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) may be accomplished by dissolving or suspending the one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in an oil vehicle. Injectable depot forms are made by forming microencapsule matrices of the one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in biodegradable polymers such as polylactide-polyglycolide. Depending upon the ratio of the one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a

pharmaceutically acceptable salt thereof) to polymer and the nature of the particular polymer employed, the rate of release for the one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable compositions or depot injectable pharmaceutical compositions may also be prepared by entrapping the one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a

pharmaceutically acceptable salt thereof) in liposomes or microemulsions that are compatible with body tissues.

[001155] Compositions or pharmaceutical compositions for rectal or vaginal administration may be suppositories that can be prepared by mixing the one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) with suitable non-irritating carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof).

[001156] The one or more cannabinoids or cannabinoid derivatives, or

pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) can be also formulated as a suppository that can be prepared from fatty emulsions or suspensions; using polyalkylene glycols such as propylene glycol, as the carrier.

[001157] Dosage forms for topical or transdermal administration of one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants, oils, or patches. The one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) are admixed under sterile conditions with an acceptable or pharmaceutically acceptable carrier and any needed preservatives or buffers as may be required. Ophthalmic compositions or ophthalmic pharmaceutical compositions, ear drops, and the like are also contemplated as being within the scope of this disclosure.

[001158] The ointments, pastes, creams, lotions, gels, solutions, inhalants, or oils may contain, in addition to one or more cannabinoids or cannabinoid derivatives, or

pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof), excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

[001159] One or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a

pharmaceutically acceptable salt thereof) may also be formulated for use as topical powders and sprays that can contain, in addition to one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof), excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. Sprays can additionally contain customary propellants such as chlorofluorohydrocarbons. [001160] Transdermal patches have the added advantage of providing controlled delivery of one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a

pharmaceutically acceptable salt thereof) to the body. Such dosage forms can be made by dissolving or dispensing the one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the proper medium. Absorption enhancers can also be used to increase the flux of one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) across the skin. The rate can be controlled by either providing a rate controlling membrane or by dispersing the one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in a polymer matrix or gel.

[001161] Compositions or pharmaceutical compositions of the disclosure may also be formulated for delivery as a liquid aerosol or inhalable dry powder. Liquid aerosol compositions or liquid aerosol pharmaceutical compositions may be nebulized

predominantly into particle sizes that can be delivered to the terminal and respiratory bronchioles. Liquid aerosol and inhalable dry powder compositions or pharmaceutical compositions may be delivered throughout the endobronchial tree to the terminal bronchioles and eventually to the parenchymal tissue.

[001162] Aerosolized compositions or aerosolized pharmaceutical compositions of the disclosure may be delivered using an aerosol forming device, such as a jet, vibrating porous plate or ultrasonic nebulizer, selected to allow the formation of an aerosol particles having with a mass medium average diameter predominantly between 1 to 5 pm. Further, the composition or pharmaceutical composition may have balanced osmolarity ionic strength and chloride concentration, and the smallest aerosolizable volume able to deliver a dose of the one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof). Additionally, the aerosolized composition or aerosolized pharmaceutical composition may not impair negatively the functionality of the airways and does not cause undesirable side effects.

[001163] Aerosolization devices suitable for administration of aerosol compositions or aerosol pharmaceutical compositions of the disclosure include, for example, jet, vibrating porous plate, ultrasonic nebulizers and energized dry powder inhalers, that are able to nebulize the composition or pharmaceutical composition of the disclosure into aerosol particle size predominantly in the size range from 1-5 pm. Predominantly in this application means that at least 70% but optionally more than 90% of all generated aerosol particles are 1 to 5 pm range. A jet nebulizer works by air pressure to break a liquid solution into aerosol droplets. Vibrating porous plate nebulizers work by using a sonic vacuum produced by a rapidly vibrating porous plate to extrude a solvent droplet through a porous plate. An ultrasonic nebulizer works by a piezoelectric crystal that shears a liquid into small aerosol droplets. A variety of suitable devices are available, including, for example, AeroNeb and AeroDose vibrating porous plate nebulizers (AeroGen, Inc., Sunnyvale, Calif), Sidestream7 nebulizers (Medic- Aid Ltd., West Sussex, England), Pari LC7 and Pari LC Star7jet nebulizers (Pari Respiratory Equipment, Inc., Richmond, Va.), and Aerosonic (DeVilbiss Medizinische Produkte (Deutschland) GmbH, Heiden, Germany) and pLtraAire7 (Omron Healthcare, Inc., Vernon Hills, Ill.) ultrasonic nebulizers.

[001164] Compositions or pharmaceutical compositions of the disclosure may also be formulated into a vaporizable formulation. In certain such embodiments, the compositions or pharmaceutical compositions of the disclosure may be delivered via direct vaporization or via an electronic inhalation device.

[001165] In some embodiments, the composition of the disclosure is an edible composition comprising one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof). “Edible composition” may refer to a composition suitable for consumption, typically via the oral cavity (although consumption may occur via non-oral means such as inhalation). Edible compositions may be present in any form including, but not limited to, liquids, solids, semi-solids, tablets, lozenges, powders, gels, gums, pastes, slurries, syrups, aerosols, and sprays. As used herein, edible compositions may include food products, pharmaceutical compositions, and consumer products. Edible compositions may also refer to, for example, dietary and nutritional supplements. As used herein, edible compositions may also include compositions that are placed within the oral cavity but not swallowed.

[001166] “Food product” may refer to any compositions comprising one or more processed foodstuff. Food products include, but are not limited to, confectionaries, bakery products, ice creams, dairy products, cheeses, sweet and savory snacks, snack foods, beverages (including, but not limited to, hot and cold beverages, beverage mixes, concentrates, juices, carbonated beverages, non-carbonated beverages, alcoholic beverages, non-alcoholic beverages, soft drinks, sports drinks, isotonic drinks, coffees, teas, bottled waters, and beverages prepared from botanicals and botanical extracts), snack bars, meal replacement products, ready meals (including, but not limited to canned meals, preserved meals, frozen meals, dried meals, chilled meals, dinner mixes, and prepared salads), soups, broth, prepared foods (including, but not limited to, dried, canned, or jarred sauces and soups), canned foods, frozen foods, dried foods, chilled foods, oils and fats, sauces, jellies, jams, preserves, honey, puddings, recipe mixes, syrups, icings, fillings, infused foods, and condiments. In some embodiments, the food product is animal feed. For example, the food product may be a pet food product, i.e. a food product for consumption by a household pet. In other embodiments, the food product is a livestock food product, i.e. a food product for consumption by livestock.

[001167] “Foodstuff’ may refer to an unprocessed ingredient or a basic nutrient or flavor containing element used to prepare a food product. Non-limiting examples of foodstuffs include: fruits, vegetables, meats, fishes, grains, milks, eggs, tubers, sugars, sweeteners, oils, herbs, snacks, sauces, spices and salts.

[001168] In some embodiments, the composition of the disclosure comprising one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) may be a consumer product.“Consumer products” may refer to health, beauty, and general wellness products for the personal use and/or consumption by a subject. Consumer products may be present in any form including, but not limited to, liquids, solids, semi solids, tablets, capsules, lozenges, strips, powders, gels, gums, pastes, slurries, syrups, aerosols and sprays. Non-limiting examples of consumer products include nutraceuticals, nutritional supplements, cosmetics, sunscreens, lotions, creams, wipes, lipsticks, lip balms, soaps, shampoos, gums, dissolvable films, adhesives (e.g., dental adhesives), toothpastes, breath fresheners, mouthwashes, and other dentifrices.

Methods of Treatment and Uses of Cannabinoids and Cannabinoid Derivatives

[001169] The cannabinoids and cannabinoid derivatives, or pharmaceutically acceptable salts thereof, described herein, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) and the pharmaceutical compositions of the disclosure may be useful as pharmaceuticals, as discussed herein. [001170] In some embodiments, one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) are useful as pharmaceuticals.

[001171] In some embodiments, one or more compounds of Formula (I), or pharmaceutically acceptable salts thereof, are useful as pharmaceuticals.

[001172] In some embodiments, the pharmaceutical composition useful as a pharmaceutical comprises one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof).

[001173] In some embodiments, the pharmaceutical composition useful as a pharmaceutical comprises one or more compounds of Formula (I), or pharmaceutically acceptable salts thereof.

[001174] The present disclosure provides a method for treating a disorder in a subject including the step of administering to a subject in need thereof a therapeutically effective amount of one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a

pharmaceutically acceptable salt thereof).

[001175] The present disclosure provides a method for treating a disorder in a subject including the step of administering to a subject in need thereof a therapeutically effective amount of one or more compounds of Formula (I), or pharmaceutically acceptable salts thereof.

[001176] The present disclosure provides a method for treating a disorder in a subject including the step of administering to a subject in need thereof a therapeutically effective amount of one or more pharmaceutical compositions of the present disclosure. In some embodiments, the present disclosure provides a method for treating a disorder in a subject including the step of administering to a subject in need thereof a therapeutically effective amount of one or more pharmaceutical compositions comprising one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof). In some embodiments, the present disclosure provides a method for treating a disorder in a subject including the step of administering to a subject in need thereof a therapeutically effective amount of one or more pharmaceutical compositions comprising one or more compounds of Formula (I), or pharmaceutically acceptable salts thereof. [001177] A“subject” may encompass both mammals and non-mammals. Examples of mammals may include, but are not limited to, any member of the class Mammalia: humans; non-human primates such as chimpanzees, monkeys, baboons, or rhesus monkeys, as well as other apes and monkey species; livestock or farm animals such as cattle, horses, sheep, goats, and swine; household pets or companion animals such as rabbits, dogs, and cats; laboratory animals including rodents, such as rats, mice and guinea pigs; and the like.

Examples of non-mammals include, but are not limited to, birds, fish, and the like.“Subject” may include both humans and animals. In some embodiments, the subject is a human.

[001178] The terms“effective amount” or“therapeutically effective amount” when used in connection with one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or pharmaceutical compositions disclosed herein may refer to a sufficient amount of the one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or pharmaceutical

compositions to provide the desired biological result. That result can be reduction and/or alleviation of the signs, symptoms, or causes of a disorder, or any other desired alteration of a biological system. For example, a“therapeutically effective amount” or“effective amount” for therapeutic use may be the amount of the pharmaceutical composition comprising one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) required to provide a clinically significant decrease in a disorder. An appropriate “therapeutically effective amount” or“effective amount” in any individual case may be determined by one of ordinary skill in the art using routine experimentation.

[001179] As used herein, the terms“treat” or“treatment” are meant to indicate a postponement of development of disorders; and/or reducing severity of such symptoms that will or are expected to develop. Thus, these terms may include ameliorating existing disorder symptoms; preventing additional symptoms; ameliorating or preventing the underlying causes of symptoms; inhibiting the disorder, e.g., arresting the development of the disorder; relieving the disorder; causing regression of the disorder; relieving a symptom caused by the disorder; or stopping or alleviating the symptoms of the disorder.

[001180] The terms“administered,”“administration,” or“administering” as used in this disclosure may refer to either directly administering one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or compositions of the disclosure to a subject.

[001181] The present disclosure provides for use of one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the

manufacture of a medicament for treating a disorder in a subject in need thereof.

[001182] The present disclosure provides for use of one or more compounds of Formula (I), or pharmaceutically acceptable salts thereof, in the manufacture of a medicament for treating a disorder in a subject in need thereof.

[001183] The present disclosure provides for use of one or more pharmaceutical compositions of the present disclosure in the manufacture of a medicament for treating a disorder in a subject in need thereof. In some embodiments, the present disclosure provides for use of one or more pharmaceutical compositions comprising one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the

manufacture of a medicament for treating a disorder in a subject in need thereof. In some embodiments, the present disclosure provides for use of one or more pharmaceutical compositions comprising one or more compounds of Formula (I), or pharmaceutically acceptable salts thereof, in the manufacture of a medicament for treating a disorder in a subj ect in need thereof.

[001184] The present disclosure provides for use of one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) for treating a disorder in a subject in need thereof.

[001185] The present disclosure provides for use of one or more compounds of Formula (I), or pharmaceutically acceptable salts thereof, for treating a disorder in a subject in need thereof.

[001186] The present disclosure provides for use of one or more pharmaceutical compositions of the present disclosure for treating a disorder in a subject in need thereof. In some embodiments, the present disclosure provides for use of one or more pharmaceutical compositions comprising one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) for treating a disorder in a subject in need thereof. In some embodiments, the present disclosure provides for use of one or more pharmaceutical compositions comprising one or more compounds of Formula (I), or pharmaceutically acceptable salts thereof, for treating a disorder in a subject in need thereof.

[001187] The present disclosure provides one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof), for use in a method of treating a disorder in a subject in need thereof.

[001188] The present disclosure provides one or more compounds of Formula (I), or pharmaceutically acceptable salts thereof, for use in a method of treating a disorder in a subject in need thereof.

[001189] The present disclosure provides one or more pharmaceutical compositions of the present disclosure for use in a method of treating a disorder in a subject in need thereof. In some embodiments, the present disclosure provides one or more pharmaceutical compositions comprising one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) for use in a method of treating a disorder in a subject in need thereof. In some embodiments, the present disclosure provides one or more pharmaceutical compositions comprising one or more compounds of Formula (I), or pharmaceutically acceptable salts thereof, for use in a method of treating a disorder in a subject in need thereof.

[001190] In some embodiments, the disorder of the methods, uses, compounds for use, medicaments, or pharmaceutical compositions for use of the disclosure is mediated by cannabinoid receptor type 1 or cannabinoid receptor type 2. In some embodiments, the disorder is mediated by cannabinoid receptor type 1. In some embodiments, the disorder is mediated by cannabinoid receptor type 2.

[001191] In some embodiments, the disorder of the methods, uses, compounds for use, medicaments, or pharmaceutical compositions for use of the disclosure is chronic pain, multiple sclerosis, cancer-associated nausea and vomiting, weight loss, appetite loss, spasticity, or seizures. In some embodiments, the disorder is chronic pain. In some embodiments, the disorder is multiple sclerosis. In some embodiments, the disorder is cancer-associated nausea and vomiting. In some embodiments, the disorder is weight loss. In some embodiments, the disorder is appetite loss. In some embodiments, the disorder is spasticity. In some embodiments, the disorder is seizures.

[001192] In some embodiments, the disorder of the methods, uses, compounds for use, medicaments, or pharmaceutical compositions for use of the disclosure is arthritis, depression, rheumatoid arthritis, osteoarthritis, nausea, alcohol use disorders, dermatitis, eczema, acne vulgaris, asthma, angina, cardiovascular disease, autoimmune disease, immunodeficiency disease, an inflammatory disorder, gastritis, epilepsy, encephalopathy, neuropathic pain, cancer (e.g., colorectal), psoriasis, Huntington’s disease, anorexia, bladder dysfunction, anxiety, opioid use disorder, dry skin syndrome, autism, Parkinson’s disease, obesity, diabetes, or inflammatory bowel disease.

[001193] For the therapeutic methods, uses, compounds for use, medicaments, and pharmaceutical compositions for use mentioned herein, the dosage administered will, of course, vary with the one or more cannabinoids or cannabinoid derivatives, or

pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or pharmaceutical compositions of the disclosure employed, the mode of administration, the treatment desired and the disorder indicated.

[001194] It will be understood, however, that the total daily usage of the one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or pharmaceutical compositions of the present disclosure will be decided by the attending physician or veterinarian within the scope of sound medical judgment. The specific therapeutically effective dose level for any particular subject will depend upon a variety of factors including the disorder being treated and the severity of the disorder; the activity of the one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a

pharmaceutically acceptable salt thereof) or pharmaceutical compositions of the present disclosure employed; the specific pharmaceutical composition employed; the age, body weight, general health, sex and diet of the subject; the time of administration, route of administration, and rate of excretion of the one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or pharmaceutical

compositions of the present disclosure employed; the duration of the treatment; drugs used in combination or coincidental with the one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or pharmaceutical compositions of the present disclosure employed; and like factors well known in the medical arts. A physician or veterinarian of ordinary skill in the art can readily determine and prescribe the therapeutically effective amount of the one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or pharmaceutical compositions disclosed herein required to treat, counter, or arrest the progress of the disorder.

[001195] In some embodiments, one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof), or compositions (including GRAS and/or pharmaceutical compositions) described herein, are useful as a nutraceutical. In some embodiments, the nutraceutical is useful for sport or athletic recovery. As used herein,“sport or athletic recovery” may include, but is not limited to, muscle repair and recovery and alleviation or treatment of soreness, swelling, pain, and inflammation associated with participation in sports, athletics, training, and/or exercise.

[001196] In some embodiments, one or more compounds of Formula (I), or pharmaceutically acceptable salts thereof, or compositions (including GRAS and/or pharmaceutical compositions) described herein, are useful as a nutraceutical. In some embodiments, the nutraceutical is useful for sport or athletic recovery.

[001197] The present disclosure provides for use of one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof), or compositions (including GRAS and/or pharmaceutical compositions) described herein, in the manufacture of a nutraceutical. In some embodiments, the nutraceutical is useful for sport or athletic recovery.

[001198] The present disclosure provides for use of one or more compounds of Formula (I), or pharmaceutically acceptable salts thereof, or compositions (including GRAS and/or pharmaceutical compositions) described herein, in the manufacture of a nutraceutical. In some embodiments, the nutraceutical is useful for sport or athletic recovery.

[001199] The present disclosure provides for use of one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof), or compositions (including GRAS and/or pharmaceutical compositions) described herein, as a nutraceutical. In some embodiments, the nutraceutical is useful for sport or athletic recovery.

[001200] The present disclosure provides for use of one or more compounds of Formula (I), or pharmaceutically acceptable salts thereof, or compositions (including GRAS and/or pharmaceutical compositions) described herein, as a nutraceutical. In some embodiments, the nutraceutical is useful for sport or athletic recovery.

[001201] The present disclosure provides for use of one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof), or compositions (including GRAS and/or pharmaceutical compositions) described herein, in the manufacture of an edible composition. In certain such embodiments, the edible composition is a food product or a consumer product. In some embodiments, the food product or consumer product is useful for sport or athletic recovery.

[001202] The present disclosure provides for use of one or more compounds of Formula (I), or pharmaceutically acceptable salts thereof, or compositions (including GRAS and/or pharmaceutical compositions) described herein, in the manufacture of an edible composition. In certain such embodiments, the edible composition is a food product or consumer product. In some embodiments, the food product or a consumer product is useful for sport or athletic recovery.

[001203] The present disclosure provides for use of one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof), or compositions (including GRAS and/or pharmaceutical compositions) described herein, in an edible composition. In certain such embodiments, the edible composition is a food product or consumer product. In some embodiments, the food product or a consumer product is useful for sport or athletic recovery.

[001204] The present disclosure provides for use of one or more compounds of Formula (I), or pharmaceutically acceptable salts thereof, or compositions (including GRAS and/or pharmaceutical compositions) described herein, in an edible composition. In certain such embodiments, the edible composition is a food product or a consumer product. In some embodiments, the food product or consumer product is useful for sport or athletic recovery.

[001205] The present disclosure provides for use of one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof), or compositions (including GRAS and/or pharmaceutical compositions) described herein, in the manufacture of a consumer product. In some embodiments, the consumer product is useful for sport or athletic recovery. [001206] The present disclosure provides for use of one or more compounds of Formula (I), or pharmaceutically acceptable salts thereof, or compositions (including GRAS and/or pharmaceutical compositions) described herein, in the manufacture of a consumer product. In some embodiments, the consumer product is useful for sport or athletic recovery.

[001207] The present disclosure provides for use of one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof), or compositions (including GRAS and/or pharmaceutical compositions) described herein, in a consumer product. In some embodiments, the consumer product is useful for sport or athletic recovery.

[001208] The present disclosure provides for use of one or more compounds of Formula (I), or pharmaceutically acceptable salts thereof, or compositions (including GRAS and/or pharmaceutical compositions) described herein, in a consumer product. In some embodiments, the consumer product is useful for sport or athletic recovery.

[001209] The present disclosure provides for use of one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof), or compositions (including GRAS and/or pharmaceutical compositions) described herein, in the manufacture of a food product. In some embodiments, the food product is useful for sport or athletic recovery.

[001210] The present disclosure provides for use of one or more compounds of Formula (I), or pharmaceutically acceptable salts thereof, or compositions (including GRAS and/or pharmaceutical compositions) described herein, in the manufacture of a food product. In some embodiments, the food product is useful for sport or athletic recovery.

[001211] The present disclosure provides for use of one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof), or compositions (including GRAS and/or pharmaceutical compositions) described herein, in a food product. In some embodiments, the food product is useful for sport or athletic recovery.

[001212] The present disclosure provides for use of one or more compounds of Formula (I), or pharmaceutically acceptable salts thereof, or compositions (including GRAS and/or pharmaceutical compositions) described herein, in a food product. In some embodiments, the food product is useful for sport or athletic recovery.

[001213] In some embodiments, the methods, uses, compounds for use, compositions for use, medicaments, or pharmaceutical compositions for use of the disclosure comprise one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof). In some embodiments, the methods, uses, compounds for use, compositions for use, medicaments, or pharmaceutical compositions for use of the disclosure comprise a cannabinoid or cannabinoid derivative, or a pharmaceutically acceptable salt thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof). In some embodiments, the methods, uses, compounds for use, compositions for use, medicaments, or pharmaceutical compositions for use of the disclosure comprise two or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof). In some embodiments, the methods, uses, compounds for use, compositions for use, medicaments, or pharmaceutical compositions for use of the disclosure comprise three or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof). In some embodiments, the methods, uses, compounds for use, compositions for use, medicaments, or pharmaceutical compositions for use of the disclosure comprise four or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof). In some embodiments, the methods, uses, compounds for use, compositions for use, medicaments, or pharmaceutical compositions for use of the disclosure comprise five or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof).

[001214] In some embodiments, the methods, uses, compounds for use, compositions for use, medicaments, or pharmaceutical compositions for use of the disclosure comprise one or more compounds of Formula (I), or pharmaceutically acceptable salts thereof. In some embodiments, the methods, uses, compounds for use, compositions for use, medicaments, or pharmaceutical compositions for use of the disclosure comprise a compound of Formula (I), or a pharmaceutically acceptable salt thereof. In some embodiments, the methods, uses, compounds for use, compositions for use, medicaments, or pharmaceutical compositions for use of the disclosure comprise two or more compounds of Formula (I), or pharmaceutically acceptable salts thereof. In some embodiments, the methods, uses, compounds for use, compositions for use, medicaments, or pharmaceutical compositions for use of the disclosure comprise three or more compounds of Formula (I), or pharmaceutically acceptable salts thereof. In some embodiments, the methods, uses, compounds for use, compositions for use, medicaments, or pharmaceutical compositions for use of the disclosure comprise four or more compounds of Formula (I), or pharmaceutically acceptable salts thereof. In some embodiments, the methods, uses, compounds for use, compositions for use, medicaments, or pharmaceutical compositions for use of the disclosure comprise five or more compounds of Formula (I), or pharmaceutically acceptable salts thereof.

Combination Therapy

[001215] In some embodiments, one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof), compositions, or pharmaceutical compositions described herein may be used alone or together or conjointly administered, or used in combination, with a known therapeutic agent, composition, or pharmaceutical composition. Conjoint administration or used in combination may refer to any form of administration of two or more different compounds, compositions, or pharmaceutical compositions such that the second compound, composition, or pharmaceutical composition is administered while the previously administered compound, composition, or

pharmaceutical composition is still active in the body. For example, the different compounds, compositions, or pharmaceutical compositions can be administered either in the same formulation or in a separate formulation, either simultaneously, sequentially, or by separate dosing of the individual components of the treatment. In some embodiments, the different compounds, compositions, or pharmaceutical compositions can be administered within about one hour, about 12 hours, about 24 hours, about 36 hours, about 48 hours, about 72 hours, or about a week of one another. Thus, a subject who receives such treatment can benefit from a combined effect of different compounds, compositions, or pharmaceutical compositions.

[001216] In some embodiments, one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof), compositions, or pharmaceutical compositions of the disclosure are used in combination with one or more other cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (1), or a pharmaceutically acceptable salt thereof),

compositions, or pharmaceutical compositions of the disclosure in the methods, uses, compounds for use, compositions for use, medicaments, or pharmaceutical compositions for use of the disclosure. In certain such embodiments, the combination of one or more other cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof), compositions, or pharmaceutical compositions of the disclosure is used in a method for treating one or more of the disorders listed herein.

[001217] In some embodiments, combinations of one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof), compositions, or pharmaceutical compositions provided herein, or combinations of other known agents, compositions, or pharmaceutical compositions and one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof), compositions, or

pharmaceutical compositions provided herein, are formulated into compositions, pharmaceutical compositions, and medicaments that are useful in the methods, uses, compounds for use, compositions for use, medicaments, or pharmaceutical compositions for use of the disclosure. The disclosure also provides for use of such combinations in treating one or more of the disorders listed herein.

[001218] In some embodiments of the disclosure, one or more cannabinoids or cannabinoid derivatives, or pharmaceutically acceptable salts thereof, disclosed herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof), compositions, or pharmaceutical compositions of the disclosure are administered at a sub-therapeutic dose, wherein a subtherapeutic dose is a dose that would be insufficient to treat one of the disorders listed herein if administered alone.

EXAMPLES OF NON-LIMITING EMBODIMENTS OF THE DISCLOSURE

[001219] Embodiments of the present subject matter disclosed herein may be beneficial alone or in combination with one or more other embodiments. Without limiting the foregoing description, certain non-limiting embodiments of the disclosure, numbered 1-1 to 1-201, II- 1 to 11-201, III-1 to III-26, and IV- 1 to IV- 17, are provided below. As will be apparent to those of skill in the art upon reading this disclosure, each of the individually numbered embodiments may be used or combined with any of the preceding or following individually numbered embodiments. This is intended to provide support for all such combinations of embodiments and is not limited to combinations of embodiments explicitly provided below.

[001220] Some embodiments of the disclosure are of Embodiment I: [001221] Embodiment 1-1. A nucleic acid comprising one or more codon- optimized nucleotide sequences having at least 80% sequence identity to SEQ ID NO:2,

SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8.

[001222] Embodiment 1-2. The nucleic acid of Embodiment 1-1, comprising one or more codon-optimized nucleotide sequences having at least 85% sequence identity to SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8.

[001223] Embodiment 1-3. The nucleic acid of Embodiment 1-1, comprising one or more codon-optimized nucleotide sequences having at least 90% sequence identity to SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8.

[001224] Embodiment 1-4. The nucleic acid of Embodiment 1-1, comprising one or more codon-optimized nucleotide sequences having at least 95% sequence identity to SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8.

[001225] Embodiment 1-5. A nucleic acid comprising one or more codon- optimized nucleotide sequences having at least 80% sequence identity to SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO:263.

[001226] Embodiment 1-6 The nucleic acid of Embodiment 1-5, comprising one or more codon-optimized nucleotide sequences having at least 85% sequence identity to SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO: 263

[001227] Embodiment 1-7. The nucleic acid of Embodiment 1-5, comprising one or more codon-optimized nucleotide sequences having at least 90% sequence identity to SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO: 263.

[001228] Embodiment 1-8. The nucleic acid of Embodiment 1-5, comprising one or more codon-optimized nucleotide sequences having at least 95% sequence identity to SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO: 263.

[001229] Embodiment 1-9. A nucleic acid comprising one or more codon- optimized nucleotide sequences having at least 80% sequence identity to SEQ ID NO:238, SEQ ID NO:27l, SEQ ID NO:272, SEQ ID NO:273, or SEQ ID NO:274. [001230] Embodiment I- 10. The nucleic acid of Embodiment 1-9, comprising one or more codon-optimized nucleotide sequences having at least 85% sequence identity to SEQ ID NO:238, SEQ ID NO:271, SEQ ID NO:272, SEQ ID NO:273, or SEQ ID NO:274.

[001231] Embodiment 1-11. The nucleic acid of Embodiment 1-9, comprising one or more codon-optimized nucleotide sequences having at least 90% sequence identity to SEQ ID NO:238, SEQ ID NO:271, SEQ ID NO:272, SEQ ID NO:273, or SEQ ID NO:274.

[001232] Embodiment 1-12. The nucleic acid of Embodiment 1-9, comprising one or more codon-optimized nucleotide sequences having at least 95% sequence identity to SEQ ID NO:238, SEQ ID NO:271, SEQ ID NO:272, SEQ ID NO:273, or SEQ ID NO:274.

[001233] Embodiment 1-13. A nucleic acid comprising one or more codon- optimized nucleotide sequences having at least 80% sequence identity to SEQ ID NO:240.

[001234] Embodiment 1-14. The nucleic acid of Embodiment 1-13, comprising one or more codon-optimized nucleotide sequences having at least 85% sequence identity to SEQ ID NO:240.

[001235] Embodiment 1-15. The nucleic acid of Embodiment 1-13, comprising one or more codon-optimized nucleotide sequences having at least 90% sequence identity to SEQ ID NO:240.

[001236] Embodiment 1-16. The nucleic acid of Embodiment 1-13, comprising one or more codon-optimized nucleotide sequences having at least 95% sequence identity to SEQ ID NO:240.

[001237] Embodiment 1-17. A nucleic acid comprising one or more codon- optimized nucleotide sequences having at least 80% sequence identity to SEQ ID NO:244.

[001238] Embodiment 1-18. The nucleic acid of Embodiment 1-17, comprising one or more codon-optimized nucleotide sequences having at least 85% sequence identity to SEQ ID NO:244.

[001239] Embodiment 1-19. The nucleic acid of Embodiment 1-17, comprising one or more codon-optimized nucleotide sequences having at least 90% sequence identity to SEQ ID NO:244.

[001240] Embodiment 1-20. The nucleic acid of Embodiment 1-17, comprising one or more codon-optimized nucleotide sequences having at least 95% sequence identity to SEQ ID NO:244.

[001241] Embodiment 1-21. The nucleic acid of any one of Embodiments 1-17 to I-

20, further comprising one or more nucleotide sequences encoding one or more signal sequence polypeptides. [001242] Embodiment 1-22. The nucleic acid of Embodiment 1-21, wherein the one or more signal sequence polypeptides comprise a secretory signal sequence polypeptide.

[001243] Embodiment 1-23. The nucleic acid of Embodiment 1-22, wherein the secretory signal sequence polypeptide is a native secretory signal sequence polypeptide.

[001244] Embodiment 1-24. The nucleic acid of Embodiment 1-22, wherein the secretory signal sequence polypeptide is a synthetic secretory signal sequence polypeptide.

[001245] Embodiment 1-25. The nucleic acid of Embodiment 1-22, wherein the secretory signal sequence polypeptide is an endoplasmic reticulum retention signal sequence polypeptide.

[001246] Embodiment 1-26. The nucleic acid of Embodiment 1-22, wherein the secretory signal sequence polypeptide is a vacuolar localization signal sequence polypeptide.

[001247] Embodiment 1-27. The nucleic acid of Embodiment 1-26, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide or a PRClt polypeptide.

[001248] Embodiment 1-28. The nucleic acid of Embodiment 1-27, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide.

[001249] Embodiment 1-29. The nucleic acid of Embodiment 1-22, wherein the secretory signal sequence polypeptide is a plasma membrane localization signal sequence polypeptide.

[001250] Embodiment 1-30. The nucleic acid of Embodiment 1-22, wherein the secretory signal sequence polypeptide is a peroxisome targeting signal sequence polypeptide.

[001251] Embodiment 1-31. A method of making a modified host cell for producing a cannabinoid or cannabinoid derivative, comprising introducing one or more nucleic acids of any one of Embodiments 1-1 to 1-30 into a host cell.

[001252] Embodiment 1-32. A vector comprising one or more nucleic acids of any one of Embodiments 1-1 to 1-30.

[001253] Embodiment 1-33. A method of making a modified host cell for producing a cannabinoid or a cannabinoid derivative, comprising introducing one or more vectors of Embodiment 1-32 into a host cell.

[001254] Embodiment 1-34. A modified host cell for producing a cannabinoid or cannabinoid derivative comprising one or more nucleic acids of any one of Embodiments 1-1 to 1-30.

[001255] Embodiment 1-35. The modified host cell of Embodiment 1-34, wherein the modified host cell is a eukaryotic cell. [001256] Embodiment 1-36. The modified host cell of Embodiment 1-35, wherein the eukaryotic cell is a yeast cell.

[001257] Embodiment 1-37. The modified host cell of Embodiment 1-36, wherein the yeast cell is Saccharomyces cerevisiae.

[001258] Embodiment 1-38. The modified host cell of Embodiment 1-37, wherein the Saccharomyces cerevisiae is a protease-deficient strain of Saccharomyces cerevisiae.

[001259] Embodiment 1-39. The modified host cell of any one of Embodiments 1-34 to 1-38, wherein at least one of the one or more nucleic acids are integrated into the chromosome of the modified host cell.

[001260] Embodiment 1-40. The modified host cell of any one of Embodiments 1-34 to 1-38, wherein at least one of the one or more nucleic acids are maintained

extrachromosomally.

[001261] Embodiment 1-41. The modified host cell of any one of Embodiments 1-34 to 1-40, wherein at least one of the one or more nucleic acids are operably-linked to an inducible promoter.

[001262] Embodiment 1-42. The modified host cell of any one of Embodiments 1-34 to 1-40, wherein at least one of the one or more nucleic acids are operably-linked to a constitutive promoter.

[001263] Embodiment 1-43. A modified host cell for producing a cannabinoid or cannabinoid derivative, wherein the modified host cell comprises one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more secretory pathway polypeptides or a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, or a combination of any of the foregoing, and wherein the modified host cell comprises one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a cannabinoid synthase polypeptide.

[001264] Embodiment 1-44. The modified host cell of Embodiment 1-43, wherein the modified host cell comprises one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more secretory pathway polypeptides.

[001265] Embodiment 1-45. The modified host cell of Embodiment 1-44, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more flavin mononucleotide (FMN) synthetase polypeptides, one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more glycosidase polypeptides, one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more

glycosyltransferase polypeptides, one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more peptidyl-prolyl isomerase polypeptides, one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more thiol oxidase polypeptides, one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more fatty acid desaturase polypeptides, one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more protein transport and trafficking polypeptides, one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more signal peptidase or signal peptidase complex polypeptides, one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR), one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more polypeptides involved in endoplasmic reticulum-associated degradation (ERAD), one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum, one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more polypeptides involved in cell wall assembly, or one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more polypeptides involved in vacuolar protein sorting, or a combination of any of the foregoing.

[001266] Embodiment 1-46. The modified host cell of Embodiment 1-45, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides.

[001267] Embodiment 1-47. The modified host cell of Embodiment 1-46, wherein the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide, a JEM1 polypeptide, a LHSl polypeptide, a SIS1 polypeptide, a SSB1 polypeptide, a CNE1 polypeptide, a CNS1 polypeptide, a PFD2s polypeptide, a PFD1 polypeptide, a SSA1 polypeptide, a YDJT polypeptide, a SIL1 polypeptide, a SCJ1 polypeptide, a ROTl polypeptide, a TCP1 polypeptide, a CCT2 polypeptide, a CCT3 polypeptide, a CCT4 polypeptide, a CCT5 polypeptide, a CCT6 polypeptide, a CCT7 polypeptide, a CCT8 polypeptide, a DED1 polypeptide, a CPR5 polypeptide, or a FPR1 polypeptide, or a combination of any of the foregoing.

[001268] Embodiment 1-48. The modified host cell of Embodiment 1-47, wherein the one or more chaperone or co-chaperone polypeptides comprise a JEM1 polypeptide and a KAR2 polypeptide.

[001269] Embodiment 1-49. The modified host cell of Embodiment 1-47, wherein the one or more chaperone or co-chaperone polypeptides comprise a LHS1 polypeptide and a KAR2 polypeptide.

[001270] Embodiment 1-50. The modified host cell of Embodiment 1-47, wherein the one or more chaperone or co-chaperone polypeptides comprise a SIL1 polypeptide and a KAR2 polypeptide.

[001271] Embodiment 1-51. The modified host cell of Embodiment 1-47, wherein the one or more chaperone or co-chaperone polypeptides comprise a CNE1 polypeptide and a KAR2 polypeptide.

[001272] Embodiment 1-52. The modified host cell of Embodiment 1-47, wherein the one or more chaperone or co-chaperone polypeptides comprise a SIS1 polypeptide and a KAR2 polypeptide.

[001273] Embodiment 1-53. The modified host cell of Embodiment 1-47, wherein the one or more chaperone or co-chaperone polypeptides comprise a SSB1 polypeptide and a KAR2 polypeptide.

[001274] Embodiment 1-54. The modified host cell of Embodiment 1-47, wherein the one or more chaperone or co-chaperone polypeptides comprise a ROT1 polypeptide and a KAR2 polypeptide.

[001275] Embodiment 1-55. The modified host cell of Embodiment 1-47, wherein the one or more chaperone or co-chaperone polypeptides comprise a CPR5 polypeptide and a KAR2 polypeptide.

[001276] Embodiment 1-56. The modified host cell of Embodiment 1-47, wherein the one or more chaperone or co-chaperone polypeptides comprise a DED1 polypeptide and a KAR2 polypeptide. [001277] Embodiment 1-57. The modified host cell of Embodiment 1-47, wherein the one or more chaperone or co-chaperone polypeptides comprise a PFD1 polypeptide, a PFD2s polypeptide, and a KAR2 polypeptide.

[001278] Embodiment 1-58. The modified host cell of Embodiment 1-47, wherein the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide.

[001279] Embodiment 1-59. The modified host cell of any one of Embodiments 1-47 to 1-58, wherein the modified host cell comprises two or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a KAR2 polypeptide.

[001280] Embodiment 1-60. The modified host cell of any one of Embodiments 1-47 to 1-58, wherein the modified host cell comprises three or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a KAR2 polypeptide.

[001281] Embodiment 1-61. The modified host cell of Embodiment 1-47, wherein the one or more chaperone or co-chaperone polypeptides comprise a SSB1 polypeptide.

[001282] Embodiment 1-62. The modified host cell of any one of Embodiments 1-45 to 1-61, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more polypeptides involved in unfolded protein response.

[001283] Embodiment 1-63. The modified host cell of Embodiment 1-62, wherein the one or more polypeptides involved in unfolded protein response comprise an IRE1 polypeptide, a HACls polypeptide, a DED1 polypeptide, or a PPQ1 polypeptide, or a combination of any of the foregoing.

[001284] Embodiment 1-64. The modified host cell of Embodiment 1-63, wherein the one or more polypeptides involved in unfolded protein response comprise an IRE1 polypeptide.

[001285] Embodiment 1-65. The modified host cell of Embodiment 1-63 or

Embodiment 1-64, wherein the one or more polypeptides involved in unfolded protein response comprise a HACls polypeptide.

[001286] Embodiment 1-66. The modified host cell of any one of Embodiments 1-63 to 1-65, wherein the one or more polypeptides involved in unfolded protein response comprise a PPQ1 polypeptide.

[001287] Embodiment 1-67. The modified host cell of Embodiment 1-62, wherein the one or more polypeptides involved in unfolded protein response comprise a transcription factor polypeptide or a lumenal sensor polypeptide that binds to an unfolded protein response element.

[001288] Embodiment 1-68. The modified host cell of any one of Embodiments 1-45 to 1-67, further comprising a synthetic polypeptide that binds to an unfolded protein response element.

[001289] Embodiment 1-69. The modified host cell of any one of Embodiments 1-45 to 1-68, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum.

[001290] Embodiment 1-70. The modified host cell of Embodiment 1-69, wherein the one or more polypeptides involved in protein translocation into the endoplasmic reticulum comprise a KAR2 polypeptide, a SEC61 polypeptide, a SBH1 polypeptide, a SSS1 polypeptide, a SRP14 polypeptide, a SRP21 polypeptide, a SRP68 polypeptide, a SRP72 polypeptide, a SEC65 polypeptide, a SRP54 polypeptide, a SEC62 polypeptide, a SEC63 polypeptide, a SEC66 polypeptide, a SEC72 polypeptide, a SRP101 polypeptide, or a SRP102 polypeptide, or a combination of any of the foregoing.

[001291] Embodiment 1-71. The modified host cell of Embodiment 1-70, wherein the one or more polypeptides involved in protein translocation into the endoplasmic reticulum comprise a SEC61 polypeptide, a SBH1 polypeptide, a SSS1 polypeptide, a SRP14 polypeptide, a SRP21 polypeptide, a SRP68 polypeptide, a SRP72 polypeptide, a SEC65 polypeptide, and a SRP54 polypeptide.

[001292] Embodiment 1-72. The modified host cell of Embodiment 1-70, wherein the one or more polypeptides involved in protein translocation into the endoplasmic reticulum comprise a SRP14 polypeptide, a SRP21 polypeptide, a SRP68 polypeptide, a SRP72 polypeptide, a SEC65 polypeptide, and a SRP54 polypeptide.

[001293] Embodiment 1-73. The modified host cell of any one of Embodiments 1-69 to 1-72, further comprising one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding SRP protein complex RNA component SCR1.

[001294] Embodiment 1-74. The modified host cell of any one of Embodiments 1-45 to 1-73, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides. [001295] Embodiment 1-75. The modified host cell of Embodiment 1-74, wherein the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide.

[001296] Embodiment 1-76. The modified host cell of any one of Embodiments 1-45 to 1-75, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more flavin mononucleotide (FMN) synthetase polypeptides.

[001297] Embodiment 1-77. The modified host cell of Embodiment 1-76, wherein the one or more flavin mononucleotide (FMN) synthetase polypeptides comprise a FMN1 polypeptide.

[001298] Embodiment 1-78. The modified host cell of any one of Embodiments 1-45 to 1-77, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more glycosidase polypeptides.

[001299] Embodiment 1-79. The modified host cell of Embodiment 1-78, wherein the one or more glycosidase polypeptides comprise a CWH41 polypeptide.

[001300] Embodiment 1-80. The modified host cell of any one of Embodiments 1-45 to 1-79, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more fatty acid desaturase polypeptides.

[001301] Embodiment 1-81. The modified host cell of Embodiment 1-80, wherein the one or more fatty acid desaturase polypeptides comprise an OLE1 polypeptide.

[001302] Embodiment 1-82. The modified host cell of any one of Embodiments 1-45 to 1-81, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more protein disulfide isomerase polypeptides.

[001303] Embodiment 1-83. The modified host cell of Embodiment 1-82, wherein the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide.

[001304] Embodiment 1-84. The modified host cell of any one of Embodiments 1-45 to 1-83, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more thiol oxidase polypeptides.

[001305] Embodiment 1-85. The modified host cell of Embodiment 1-84, wherein the one or more thiol oxidase polypeptides comprise an EROl polypeptide or an ERV2 polypeptide, or a combination of any of the foregoing.

[001306] Embodiment 1-86. The modified host cell of Embodiment 1-85, wherein the one or more thiol oxidase polypeptides comprise an EROl polypeptide.

[001307] Embodiment 1-87. The modified host cell of Embodiment 1-85 or

Embodiment 1-86, wherein the modified host cell comprises two or more heterologous nucleic acids comprising one or more nucleotide sequences encoding an EROl polypeptide.

[001308] Embodiment 1-88. The modified host cell of any one of Embodiments 1-85 to 1-87, wherein the one or more thiol oxidase polypeptides comprise an ERV2 polypeptide.

[001309] Embodiment 1-89. The modified host cell of any one of Embodiments 1-45 to 1-88, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more signal peptidase or signal peptidase complex polypeptides.

[001310] Embodiment 1-90. The modified host cell of Embodiment 1-89, wherein the one or more signal peptidase or signal peptidase complex polypeptides comprise a KEX2 polypeptide, a SPC1 polypeptide, a SPC2 polypeptide, a SPC3 polypeptide, or an SEC 11 polypeptide, or a combination of any of the foregoing.

[001311] Embodiment 1-91. The modified host cell of any one of Embodiments 1-45 to 1-90, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more protein transport and trafficking polypeptides.

[001312] Embodiment 1-92. The modified host cell of Embodiment 1-91, wherein the one or more protein transport and trafficking polypeptides comprise a SEC23 polypeptide, a SEC24 polypeptide, a SEC 13 polypeptide, a BFR2 polypeptide, a SEC31 polypeptide, a SAR1 polypeptide, a SEC 12 polypeptide, a SEC4 polypeptide, a SEC 16 polypeptide, an ERV29 polypeptide, a SEC7 polypeptide, a SEC5 polypeptide, a SEC22 polypeptide, a BOSl polypeptide, a BETl polypeptide, a BElGl polypeptide, a GRH1 polypeptide, a USOl polypeptide, a SEC17 polypeptide, a SEC18 polypeptide, a SSOl polypeptide, a SEC9 polypeptide, a SNC2 polypeptide, or a LDB17 polypeptide, or a combination of any of the foregoing.

[001313] Embodiment 1-93. The modified host cell of Embodiment 1-92, wherein the one or more protein transport and trafficking polypeptides comprise a BFR2 polypeptide.

[001314] Embodiment 1-94. The modified host cell of Embodiment 1-92 or

Embodiment 1-93, wherein the one or more protein transport and trafficking polypeptides comprise a SEC24 polypeptide.

[001315] Embodiment 1-95. The modified host cell of any one of Embodiments 1-92 to 1-94, wherein the one or more protein transport and trafficking polypeptides comprise a LDB17 polypeptide.

[001316] Embodiment 1-96. The modified host cell of any one of Embodiments 1-45 to 1-95, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more polypeptides involved in endoplasmic reticulum-associated degradation (ERAD).

[001317] Embodiment 1-97. The modified host cell of Embodiment 1-96, wherein the one or more polypeptides involved in endoplasmic reticulum-associated degradation (ERAD) comprise a NPL4 polypeptide.

[001318] Embodiment 1-98. The modified host cell of any one of Embodiments 1-45 to 1-97, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more polypeptides involved in cell wall assembly.

[001319] Embodiment 1-99. The modified host cell of Embodiment 1-98, wherein the one or more polypeptides involved in cell wall assembly comprise a KRE1 polypeptide.

[001320] Embodiment I- 100. The modified host cell of any one of Embodiments 1-45 to 1-99, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more polypeptides involved in vacuolar protein sorting.

[001321] Embodiment 1-101. The modified host cell of Embodiment 1-100, wherein the one or more polypeptides involved in vacuolar protein sorting comprise a PEP1 polypeptide. [001322] Embodiment 1-102. The modified host cell of any one of Embodiments 1-45 to 1-101, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more glycosyltransferase polypeptides.

[001323] Embodiment 1-103. The modified host cell of any one of Embodiments 1-43 to 1-102, wherein the modified host cell comprises a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides.

[001324] Embodiment 1-104. The modified host cell of Embodiment 1-103, wherein the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more glycosidase polypeptides, one or more genes encoding one or more glycosyltransferase polypeptides, one or more genes encoding one or more polypeptides involved in lipid droplet assembly, one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism, or one or more genes encoding one or more vacuolar proteinase polypeptides, or a combination of any of the foregoing.

[001325] Embodiment 1-105. The modified host cell of Embodiment 1-104, wherein the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more glycosidase polypeptides.

[001326] Embodiment 1-106. The modified host cell of Embodiment 1-105, wherein the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene or a MNS1 gene, or a combination of any of the foregoing.

[001327] Embodiment 1-107. The modified host cell of Embodiment 1-106, wherein the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene.

[001328] Embodiment 1-108. The modified host cell of any one of Embodiments I- 104 to 1-107, wherein the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more glycosyltransferase polypeptides.

[001329] Embodiment 1-109. The modified host cell of Embodiment 1-108, wherein the one or more genes encoding one or more glycosyltransferase polypeptides comprise an ALG12 gene.

[001330] Embodiment 1-110. The modified host cell of any one of Embodiments I- 104 to 1-109, wherein the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more polypeptides involved in lipid droplet assembly.

[001331] Embodiment 1-111. The modified host cell of Embodiment 1-110, wherein the one or more genes encoding one or more polypeptides involved in lipid droplet assembly comprise a FLD1 gene.

[001332] Embodiment 1-112. The modified host cell of any one of Embodiments I- 104 to 1-111, wherein the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism.

[001333] Embodiment 1-113. The modified host cell of Embodiment 1-112, wherein the one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism comprise an OPI1 gene.

[001334] Embodiment 1-114. The modified host cell of any one of Embodiments I- 104 to 1-113, wherein the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides.

[001335] Embodiment 1-115. The modified host cell of Embodiment 1-114, wherein the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene, a PRB1 gene, or a PRC1 gene, or a combination of any of the foregoing.

[001336] Embodiment 1-116. The modified host cell of Embodiment 1-115, wherein the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene.

[001337] Embodiment 1-117. The modified host cell of any one of Embodiments 1-43 to 1-116, wherein the modified host cell comprises two or more heterologous nucleic acids comprising one or more nucleotide sequences encoding the cannabinoid synthase polypeptide.

[001338] Embodiment 1-118. The modified host cell of any one of Embodiments 1-43 to 1-117, wherein the one or more nucleotide sequences encoding the cannabinoid synthase polypeptide are codon-optimized.

[001339] Embodiment 1-119. The modified host cell of any one of Embodiments 1-43 to 1-118, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide. [001340] Embodiment 1-120. The modified host cell of any one of Embodiments 1-43 to 1-118, wherein the cannabinoid synthase polypeptide is a cannabichromenic acid synthase polypeptide.

[001341] Embodiment 1-121. The modified host cell of any one of Embodiments 1-43 to 1-118, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[001342] Embodiment 1-122. The modified host cell of Embodiment 1-121, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide, comprise one or more codon-optimized nucleotide sequences having at least 80% sequence identity to SEQ ID NO:242 or SEQ ID NO: 244.

[001343] Embodiment 1-123. The modified host cell of Embodiment 1-121, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide, comprise one or more codon-optimized nucleotide sequences having at least 85% sequence identity to SEQ ID NO:242 or SEQ ID NO: 244.

[001344] Embodiment 1-124. The modified host cell of Embodiment 1-121, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide, comprise one or more codon-optimized nucleotide sequences having at least 90% sequence identity to SEQ ID NO:242 or SEQ ID NO: 244.

[001345] Embodiment 1-125. The modified host cell of Embodiment 1-121, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide, comprise one or more codon-optimized nucleotide sequences having at least 95% sequence identity to SEQ ID NO:242 or SEQ ID NO: 244.

[001346] Embodiment 1-126. The modified host cell of Embodiment 1-121, wherein the cannabidiolic acid synthase polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:243 or SEQ ID NO:245. [001347] Embodiment 1-127. The modified host cell of any one of Embodiments 1-43 to 1-126, wherein the cannabinoid synthase polypeptide comprises one or more signal sequence polypeptides.

[001348] Embodiment 1-128. The modified host cell of Embodiment 1-127, wherein the one or more signal sequence polypeptides comprise a secretory signal sequence polypeptide.

[001349] Embodiment 1-129. The modified host cell of Embodiment 1-128, wherein the secretory signal sequence polypeptide is a native secretory signal sequence polypeptide.

[001350] Embodiment 1-130. The modified host cell of Embodiment 1-128, wherein the secretory signal sequence polypeptide is a synthetic secretory signal sequence polypeptide.

[001351] Embodiment 1-131. The modified host cell of Embodiment 1-128, wherein the secretory signal sequence polypeptide is an endoplasmic reticulum retention signal sequence polypeptide.

[001352] Embodiment 1-132. The modified host cell of Embodiment 1-128, wherein the secretory signal sequence polypeptide is a vacuolar localization signal sequence polypeptide.

[001353] Embodiment 1-133. The modified host cell of Embodiment 1-132, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide or a PRClt polypeptide.

[001354] Embodiment 1-134. The modified host cell of Embodiment 1-133, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide.

[001355] Embodiment 1-135. The modified host cell of Embodiment 1-128, wherein the secretory signal sequence polypeptide is a plasma membrane localization signal sequence polypeptide.

[001356] Embodiment 1-136. The modified host cell of Embodiment 1-128, wherein the secretory signal sequence polypeptide is a peroxisome targeting signal sequence polypeptide.

[001357] Embodiment 1-137. The modified host cell of Embodiment 1-121, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide, comprise one or more codon-optimized nucleotide sequences having at least 80% sequence identity to SEQ ID NO: l, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID N0:5, SEQ ID N0:6, SEQ ID N0:7, or SEQ ID NO:8.

[001358] Embodiment 1-138. The modified host cell of Embodiment 1-121, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide, comprise one or more codon-optimized nucleotide sequences having at least 85% sequence identity to SEQ ID NO: l, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8.

[001359] Embodiment 1-139. The modified host cell of Embodiment 1-121, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide, comprise one or more codon-optimized nucleotide sequences having at least 90% sequence identity to SEQ ID NO: l, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8.

[001360] Embodiment 1-140. The modified host cell of Embodiment 1-121, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide, comprise one or more codon-optimized nucleotide sequences having at least 95% sequence identity to SEQ ID NO: l, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8

[001361] Embodiment 1-141. The modified host cell of Embodiment 1-121, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide, comprise one or more codon-optimized nucleotide sequences having at least 80% sequence identity to SEQ ID NO:256, SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO 260, SEQ ID N0 261, SEQ ID NO:262, or SEQ ID NO: 263.

[001362] Embodiment 1-142. The modified host cell of Embodiment 1-121, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide, comprise one or more codon-optimized nucleotide sequences having at least 85% sequence identity to SEQ ID NO:256, SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID N0 261, SEQ ID NO:262, or SEQ ID NO: 263.

[001363] Embodiment 1-143. The modified host cell of Embodiment 1-121, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide, comprise one or more codon-optimized nucleotide sequences having at least 90% sequence identity to SEQ ID NO:256, SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID N0 261, SEQ ID NO:262, or SEQ ID NO: 263.

[001364] Embodiment 1-144. The modified host cell of Embodiment 1-121, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide, comprise one or more codon-optimized nucleotide sequences having at least 95% sequence identity to SEQ ID NO:256, SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID N0 261, SEQ ID NO:262, or SEQ ID NO: 263.

[001365] Embodiment 1-145. The modified host cell of Embodiment 1-119, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide, comprise one or more codon- optimized nucleotide sequences having at least 80% sequence identity to SEQ ID NO:238, SEQ ID NO:27l, SEQ ID NO:272, SEQ ID NO:273, or SEQ ID NO:274.

[001366] Embodiment 1-146. The modified host cell of Embodiment 1-119, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide, comprise one or more codon- optimized nucleotide sequences having at least 85% sequence identity to SEQ ID NO:238, SEQ ID NO:27l, SEQ ID NO:272, SEQ ID NO:273, or SEQ ID NO:274.

[001367] Embodiment 1-147. The modified host cell of Embodiment 1-119, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide, comprise one or more codon- optimized nucleotide sequences having at least 90% sequence identity to SEQ ID NO:238, SEQ ID NO:27l, SEQ ID NO:272, SEQ ID NO:273, or SEQ ID NO:274.

[001368] Embodiment 1-148. The modified host cell of Embodiment 1-119, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide, comprise one or more codon- optimized nucleotide sequences having at least 95% sequence identity to SEQ ID NO:238, SEQ ID NO:27l, SEQ ID NO:272, SEQ ID NO:273, or SEQ ID NO:274.

[001369] Embodiment 1-149. The modified host cell of Embodiment 1-119, wherein the tetrahydrocannabinolic acid synthase polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:239.

[001370] Embodiment 1-150. The modified host cell of Embodiment 1-120, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a cannabichromenic acid synthase polypeptide, comprise one or more codon-optimized nucleotide sequences having at least 80% sequence identity to SEQ ID NO:240.

[001371] Embodiment 1-151. The modified host cell of Embodiment 1-120, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a cannabichromenic acid synthase polypeptide, comprise one or more codon-optimized nucleotide sequences having at least 80% sequence identity to SEQ ID NO:240.

[001372] Embodiment 1-152. The modified host cell of Embodiment 1-120, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a cannabichromenic acid synthase polypeptide, comprise one or more codon-optimized nucleotide sequences having at least 90% sequence identity to SEQ ID NO:240.

[001373] Embodiment 1-153. The modified host cell of Embodiment 1-120, wherein the one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a cannabichromenic acid synthase polypeptide, comprise one or more codon-optimized nucleotide sequences having at least 95% sequence identity to SEQ ID NO:240.

[001374] Embodiment 1-154. The modified host cell of Embodiment 1-120, wherein the cannabichromenic acid synthase polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:24l . [001375] Embodiment 1-155. The modified host cell of Embodiment 1-121, wherein the cannabidiolic acid synthase polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:9.

[001376] Embodiment 1-156. The modified host cell of Embodiment 1-121, wherein the cannabidiolic acid synthase polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:257.

[001377] Embodiment 1-157. The modified host cell of any one of Embodiments 1-43 to 1-156, further comprising one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a geranyl pyrophosphate :olivetolic acid geranyltransferase (GOT) polypeptide comprising an amino acid sequence having at least 80% sequence identity to SEQ ID NO:73.

[001378] Embodiment 1-158. The modified host cell of any one of Embodiments 1-43 to 1-156, further comprising one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a GOT polypeptide comprising an amino acid sequence having at least 80% sequence identity to SEQ ID NO:70.

[001379] Embodiment 1-159. The modified host cell of any one of Embodiments 1-43 to 1-158, further comprising one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a tetraketide synthase (TKS) polypeptide and one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding an olivetolic acid cyclase (OAC) polypeptide.

[001380] Embodiment 1-160. The modified host cell of Embodiment 1-159, wherein the TKS polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:75.

[001381] Embodiment 1-161. The modified host cell of Embodiment 1-159 or Embodiment 1-160, wherein the modified host cell comprises three or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a TKS polypeptide.

[001382] Embodiment 1-162. The modified host cell of any one of Embodiments I- 159 to 1-161, wherein the OAC polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:77 or SEQ ID NO:225.

[001383] Embodiment 1-163. The modified host cell of any one of Embodiments I- 159 to 1-162, wherein the modified host cell comprises three or more heterologous nucleic acids comprising one or more nucleotide sequences encoding an OAC polypeptide. [001384] Embodiment 1-164. The modified host cell of any one of Embodiments 1-43 to 1-163, further comprising one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding an acyl-activating enzyme (AAE) polypeptide.

[001385] Embodiment 1-165. The modified host cell of Embodiment 1-164, wherein the AAE polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:79, SEQ ID NO:268, or SEQ ID NO:270.

[001386] Embodiment 1-166. The modified host cell of Embodiment 1-164 or Embodiment 1-165, wherein the modified host cell comprises two or more heterologous nucleic acids comprising one or more nucleotide sequences encoding an AAE polypeptide.

[001387] Embodiment 1-167. The modified host cell of any one of Embodiments 1-43 to 1-166, further comprising one or more of the following: a) one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a HMG-CoA synthase (HMGS) polypeptide; b) one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a truncated 3-hydroxy-3-methyl-glutaryl-CoA reductase (tHMGR) polypeptide; c) one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a mevalonate kinase (MK) polypeptide; d) one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a phosphomevalonate kinase (PMK) polypeptide; e) one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a mevalonate pyrophosphate decarboxylase (MVD1) polypeptide; or i) one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a isopentenyl diphosphate isomerase (IDI1) polypeptide.

[001388] Embodiment 1-168. The modified host cell of Embodiment 1-167, wherein the IDI1 polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 81.

[001389] Embodiment 1-169. The modified host cell of Embodiment 1-167 or Embodiment 1-168, wherein the tHMGR polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:83.

[001390] Embodiment 1-170. The modified host cell of any one of Embodiments I- 167 to 1-169, wherein the HMGS polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:85.

[001391] Embodiment 1-171. The modified host cell of any one of Embodiments I- 167 to 1-170, wherein the MK polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:95. [001392] Embodiment 1-172. The modified host cell of any one of Embodiments I- 167 to 1-171, wherein the PMK polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:93.

[001393] Embodiment 1-173. The modified host cell of any one of Embodiments I- 167 to 1-172, wherein the MVD1 polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:89.

[001394] Embodiment 1-174. The modified host cell of any one of Embodiments 1-43 to 1-173, further comprising one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding an acetoacetyl-CoA thiolase polypeptide.

[001395] Embodiment 1-175. The modified host cell of Embodiment 1-174, wherein the acetoacetyl-CoA thiolase polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:87.

[001396] Embodiment 1-176. The modified host cell of any one of Embodiments 1-43 to 1-175, further comprising one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a pyruvate decarboxylase (PDC) polypeptide.

[001397] Embodiment 1-177. The modified host cell of Embodiment 1-176, wherein the PDC polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:91.

[001398] Embodiment 1-178. The modified host cell of any one of Embodiments 1-43 to 1-177, further comprising one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a geranyl pyrophosphate synthetase (GPPS) polypeptide.

[001399] Embodiment 1-179. The modified host cell of Embodiment 1-178, wherein the GPPS polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:97.

[001400] Embodiment 1-180. The modified host cell of any one of Embodiments 1-43 to 1-179, wherein the modified host cell is a eukaryotic cell.

[001401] Embodiment 1-181. The modified host cell of Embodiment 1-180, wherein the eukaryotic cell is a yeast cell.

[001402] Embodiment 1-182. The modified host cell of Embodiment 1-181, wherein the yeast cell is Saccharomyces cerevisiae.

[001403] Embodiment 1-183. The modified host cell of Embodiment 1-182, wherein the Saccharomyces cerevisiae is a protease-deficient strain of Saccharomyces cerevisiae. [001404] Embodiment 1-184. The modified host cell of any one of Embodiments 1-43 to 1-183, wherein at least one of the one or more heterologous nucleic acids are integrated into the chromosome of the modified host cell.

[001405] Embodiment 1-185. The modified host cell of any one of Embodiments 1-43 to 1-183, wherein at least one of the one or more heterologous nucleic acids are maintained extrachromosomally.

[001406] Embodiment 1-186. The modified host cell of any one of Embodiments 1-43 to 1-185, wherein at least one of the one or more heterologous nucleic acids are operably- linked to an inducible promoter.

[001407] Embodiment 1-187. The modified host cell of any one of Embodiments 1-43 to 1-185, wherein at least one of the one or more heterologous nucleic acids are operably- linked to a constitutive promoter.

[001408] Embodiment 1-188. A method of producing a cannabinoid or a cannabinoid derivative comprising: a) culturing a modified host cell of any one of Embodiments 1-34 to I- 187 in a culture medium; and b) recovering the produced cannabinoid or cannabinoid derivative.

[001409] Embodiment 1-189. The method of Embodiment 1-188, wherein the culture medium comprises a carboxylic acid.

[001410] Embodiment 1-190. The method of Embodiment I- 189, wherein the carboxylic acid is an unsubstituted or substituted C3-C18 fatty acid.

[001411] Embodiment 1-191. The method of Embodiment 1-190, wherein the unsubstituted or substituted C3-C18 fatty acid is an unsubstituted or substituted hexanoic acid.

[001412] Embodiment 1-192. The method of Embodiment 1-188, wherein the culture medium comprises olivetolic acid or an olivetolic acid derivative.

[001413] Embodiment 1-193. The method of any one of Embodiments 1-188 to 1-192, wherein the culture medium comprises a fermentable sugar.

[001414] Embodiment 1-194. The method of any one of Embodiments 1-188 to 1-192, wherein the culture medium comprises a pretreated cellulosic feedstock.

[001415] Embodiment 1-195. The method of any one of Embodiments 1-188 to 1-192, wherein the culture medium comprises a non-fermentable carbon source.

[001416] Embodiment 1-196. The method of Embodiment I- 195, wherein the non- fermentable carbon source comprises ethanol.

[001417] Embodiment 1-197. The method of any one of Embodiments 1-188 to 1-196, wherein the cannabinoid is cannabigerolic acid, cannabigerol, A⁹-tetrahydrocannabinolic acid, A⁹-tetrahydrocannabinol, A^s-tetrahydrocannabinolic acid, A⁸-tetrahydrocannabinol, cannabidiolic acid, cannabidiol, cannabichromenic acid, cannabichromene, cannabinolic acid, cannabinol, cannabidivarinic acid, cannabidivarin, tetrahydrocannabivarinic acid, tetrahydrocannabivarin, cannabichromevarinic acid, cannabichromevarin,

cannabigerovarinic acid, cannabigerovarin, cannabicyclolic acid, cannabicyclol,

cannabielsoinic acid, cannabielsoin, cannabicitranic acid, or cannabicitran.

[001418] Embodiment 1-198. The method of any one of Embodiments 1-188 to 1-197, wherein the cannabinoid or cannabinoid derivative is produced in a recoverable amount of more than 100 mg/L culture medium.

[001419] Embodiment 1-199. A method of making a modified host cell of any one of Embodiments 1-43 to 1-187, comprising introducing into a host cell: a) one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a cannabinoid synthase polypeptide and b) one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more secretory pathway polypeptides.

[001420] Embodiment 1-200. A method of making a modified host cell of any one of Embodiments 1-43 to 1-187, comprising introducing into a host cell: a) one or more vectors comprising one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding a cannabinoid synthase polypeptide and b) one or more vectors comprising one or more heterologous nucleic acids comprising one or more nucleotide sequences encoding one or more secretory pathway polypeptides.

[001421] Embodiment 1-201. The method of Embodiment 1-199 or Embodiment I-

200, said method further comprising introducing into the host cell a deletion or

downregulation of one or more genes encoding one or more secretory pathway polypeptides.

[001422] Some embodiments of the disclosure are of Embodiment II:

[001423] Embodiment II- 1. A nucleic acid comprising a codon-optimized nucleotide sequence having at least 80% sequence identity to SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8.

[001424] Embodiment II-2. The nucleic acid of Embodiment II- 1, comprising a codon-optimized nucleotide sequence having at least 85% sequence identity to SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8.

[001425] Embodiment II-3. The nucleic acid of Embodiment II- 1, comprising a codon-optimized nucleotide sequence having at least 90% sequence identity to SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID N0:5, SEQ ID N0:6, SEQ ID N0:7, or SEQ ID NO:8.

[001426] Embodiment II-4. The nucleic acid of Embodiment II- 1, comprising a codon-optimized nucleotide sequence having at least 95% sequence identity to SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8.

[001427] Embodiment II-5. A nucleic acid comprising a codon-optimized nucleotide sequence having at least 80% sequence identity to SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID N0 262, or SEQ ID NO:263.

[001428] Embodiment II-6. The nucleic acid of Embodiment II-5, comprising a codon-optimized nucleotide sequence having at least 85% sequence identity to SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID N0 261, SEQ ID NO:262, or SEQ ID NO: 263.

[001429] Embodiment II-7. The nucleic acid of Embodiment II-5, comprising a codon-optimized nucleotide sequence having at least 90% sequence identity to SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID N0 261, SEQ ID NO:262, or SEQ ID NO: 263.

[001430] Embodiment II-8. The nucleic acid of Embodiment II-5, comprising a codon-optimized nucleotide sequence having at least 95% sequence identity to SEQ ID N0 258, SEQ ID N0 259, SEQ ID NO 260, SEQ ID NO 261, SEQ ID N0 262, or SEQ ID NO: 263.

[001431] Embodiment II-9. A nucleic acid comprising a codon-optimized nucleotide sequence having at least 80% sequence identity to SEQ ID NO:238, SEQ ID NO:27l, SEQ ID NO:272, SEQ ID NO:273, or SEQ ID NO:274.

[001432] Embodiment II- 10. The nucleic acid of Embodiment II-9, comprising a codon-optimized nucleotide sequence having at least 85% sequence identity to SEQ ID NO:238, SEQ ID NO:27l, SEQ ID NO:272, SEQ ID N0 273, or SEQ ID NO:274

[001433] Embodiment II- 11. The nucleic acid of Embodiment II-9, comprising a codon-optimized nucleotide sequence having at least 90% sequence identity to SEQ ID NO:238, SEQ ID NO:27l, SEQ ID NO:272, SEQ ID N0 273, or SEQ ID NO:274

[001434] Embodiment 11-12. The nucleic acid of Embodiment II-9, comprising a codon-optimized nucleotide sequence having at least 95% sequence identity to SEQ ID NO:238, SEQ ID NO:271, SEQ ID NO:272, SEQ ID N0 273, or SEQ ID NO:274 [001435] Embodiment 11-13. A nucleic acid comprising a codon-optimized nucleotide sequence having at least 80% sequence identity to SEQ ID NO:240.

[001436] Embodiment 11-14. The nucleic acid of Embodiment 11-13, comprising a codon-optimized nucleotide sequence having at least 85% sequence identity to SEQ ID NO: 240.

[001437] Embodiment 11-15. The nucleic acid of Embodiment 11-13, comprising a codon-optimized nucleotide sequence having at least 90% sequence identity to SEQ ID NO: 240.

[001438] Embodiment 11-16. The nucleic acid of Embodiment 11-13, comprising a codon-optimized nucleotide sequence having at least 95% sequence identity to SEQ ID NO: 240.

[001439] Embodiment 11-17. A nucleic acid comprising a codon-optimized nucleotide sequence having at least 80% sequence identity to SEQ ID NO:244.

[001440] Embodiment 11-18. The nucleic acid of Embodiment 11-17, comprising a codon-optimized nucleotide sequence having at least 85% sequence identity to SEQ ID NO: 244.

[001441] Embodiment 11-19. The nucleic acid of Embodiment 11-17, comprising a codon-optimized nucleotide sequence having at least 90% sequence identity to SEQ ID NO: 244.

[001442] Embodiment 11-20. The nucleic acid of Embodiment 11-17, comprising a codon-optimized nucleotide sequence having at least 95% sequence identity to SEQ ID NO: 244.

[001443] Embodiment 11-21. The nucleic acid of any one of Embodiments 11-17 to 11-20, wherein the nucleic acid comprises a nucleotide sequence encoding a signal sequence polypeptide.

[001444] Embodiment 11-22. The nucleic acid of Embodiment 11-21, wherein the signal sequence polypeptide is a secretory signal sequence polypeptide.

[001445] Embodiment 11-23. The nucleic acid of Embodiment 11-22, wherein the secretory signal sequence polypeptide is a native secretory signal sequence polypeptide.

[001446] Embodiment 11-24. The nucleic acid of Embodiment 11-22, wherein the secretory signal sequence polypeptide is a synthetic secretory signal sequence polypeptide.

[001447] Embodiment 11-25. The nucleic acid of Embodiment 11-22, wherein the secretory signal sequence polypeptide is an endoplasmic reticulum retention signal sequence polypeptide. [001448] Embodiment 11-26. The nucleic acid of Embodiment 11-22, wherein the secretory signal sequence polypeptide is a vacuolar localization signal sequence polypeptide.

[001449] Embodiment 11-27. The nucleic acid of Embodiment 11-26, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide or a PRClt polypeptide.

[001450] Embodiment 11-28. The nucleic acid of Embodiment 11-27, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide.

[001451] Embodiment 11-29. The nucleic acid of Embodiment 11-22, wherein the secretory signal sequence polypeptide is a plasma membrane localization signal sequence polypeptide.

[001452] Embodiment 11-30. The nucleic acid of Embodiment 11-22, wherein the secretory signal sequence polypeptide is a peroxisome targeting signal sequence polypeptide.

[001453] Embodiment II-31. A method of making a modified host cell for producing a cannabinoid or cannabinoid derivative, the method comprising introducing one or more nucleic acids of any one of Embodiments II-1 to 11-30 into a host cell.

[001454] Embodiment 11-32. A vector comprising one or more nucleic acids of any one of Embodiments II- 1 to 11-30.

[001455] Embodiment 11-33. A method of making a modified host cell for producing a cannabinoid or a cannabinoid derivative, the method comprising introducing one or more vectors of Embodiment 11-32 into a host cell.

[001456] Embodiment 11-34. A modified host cell for producing a cannabinoid or cannabinoid derivative comprising one or more nucleic acids of any one of Embodiments II- 1 to 11-30

[001457] Embodiment 11-35. The modified host cell of Embodiment 11-34, wherein the modified host cell is a eukaryotic cell.

[001458] Embodiment 11-36. The modified host cell of Embodiment 11-35, wherein the eukaryotic cell is a yeast cell.

[001459] Embodiment 11-37. The modified host cell of Embodiment 11-36, wherein the yeast cell is Saccharomyces cerevisiae.

[001460] Embodiment 11-38. The modified host cell of Embodiment 11-37, wherein the Saccharomyces cerevisiae is a protease-deficient strain of Saccharomyces cerevisiae.

[001461] Embodiment 11-39. The modified host cell of any one of Embodiments II- 34 to 11-38, wherein at least one of the one or more nucleic acids are integrated into the chromosome of the modified host cell. [001462] Embodiment 11-40. The modified host cell of any one of Embodiments II- 34 to 11-38, wherein at least one of the one or more nucleic acids are maintained

extrachromosomally.

[001463] Embodiment 11-41. The modified host cell of any one of Embodiments II- 34 to 11-40, wherein at least one of the one or more nucleic acids are operably-linked to an inducible promoter.

[001464] Embodiment 11-42. The modified host cell of any one of Embodiments II- 34 to 11-40, wherein at least one of the one or more nucleic acids are operably-linked to a constitutive promoter.

[001465] Embodiment 11-43. A modified host cell for producing a cannabinoid or cannabinoid derivative, wherein the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides or a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, or a combination of any of the foregoing, and wherein the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide.

[001466] Embodiment 11-44. The modified host cell of Embodiment 11-43, wherein the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides.

[001467] Embodiment 11-45. The modified host cell of Embodiment 11-44, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin mononucleotide (FMN) synthetase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosidase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosyltransferase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more peptidyl-prolyl isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more fatty acid desaturase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein transport and trafficking polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more signal peptidase or signal peptidase complex polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR), one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in endoplasmic reticulum-associated degradation (ERAD), one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cell wall assembly, or one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in vacuolar protein sorting, or a combination of any of the foregoing.

[001468] Embodiment 11-46. The modified host cell of Embodiment 11-45, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides.

[001469] Embodiment 11-47. The modified host cell of Embodiment 11-46, wherein the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide, a JEM1 polypeptide, a LHSl polypeptide, a SIS1 polypeptide, a SSB1 polypeptide, a CNE1 polypeptide, a CNS1 polypeptide, a PFD2s polypeptide, a PFD1 polypeptide, a SSA1 polypeptide, a YDJT polypeptide, a SIL1 polypeptide, a SCJ1 polypeptide, a ROTl polypeptide, a TCP1 polypeptide, a CCT2 polypeptide, a CCT3 polypeptide, a CCT4 polypeptide, a CCT5 polypeptide, a CCT6 polypeptide, a CCT7 polypeptide, a CCT8 polypeptide, a DED1 polypeptide, a CPR5 polypeptide, or a FPR1 polypeptide, or a combination of any of the foregoing.

[001470] Embodiment 11-48. The modified host cell of Embodiment 11-47, wherein the one or more chaperone or co-chaperone polypeptides comprise a JEM1 polypeptide and a KAR2 polypeptide.

[001471] Embodiment 11-49. The modified host cell of Embodiment 11-47, wherein the one or more chaperone or co-chaperone polypeptides comprise a LHS1 polypeptide and a KAR2 polypeptide. [001472] Embodiment 11-50. The modified host cell of Embodiment 11-47, wherein the one or more chaperone or co-chaperone polypeptides comprise a SIL1 polypeptide and a KAR2 polypeptide.

[001473] Embodiment II-51. The modified host cell of Embodiment 11-47, wherein the one or more chaperone or co-chaperone polypeptides comprise a CNE1 polypeptide and a KAR2 polypeptide.

[001474] Embodiment 11-52. The modified host cell of Embodiment 11-47, wherein the one or more chaperone or co-chaperone polypeptides comprise a SIS1 polypeptide and a KAR2 polypeptide.

[001475] Embodiment 11-53. The modified host cell of Embodiment 11-47, wherein the one or more chaperone or co-chaperone polypeptides comprise a SSB1 polypeptide and a KAR2 polypeptide.

[001476] Embodiment 11-54. The modified host cell of Embodiment 11-47, wherein the one or more chaperone or co-chaperone polypeptides comprise a ROT1 polypeptide and a KAR2 polypeptide.

[001477] Embodiment 11-55. The modified host cell of Embodiment 11-47, wherein the one or more chaperone or co-chaperone polypeptides comprise a CPR5 polypeptide and a KAR2 polypeptide.

[001478] Embodiment 11-56. The modified host cell of Embodiment 11-47, wherein the one or more chaperone or co-chaperone polypeptides comprise a DED1 polypeptide and a KAR2 polypeptide.

[001479] Embodiment 11-57. The modified host cell of Embodiment 11-47, wherein the one or more chaperone or co-chaperone polypeptides comprise a PFD1 polypeptide, a PFD2s polypeptide, and a KAR2 polypeptide.

[001480] Embodiment 11-58. The modified host cell of Embodiment 11-47, wherein the one or more chaperone or co-chaperone polypeptides comprise a KAR2 polypeptide.

[001481] Embodiment 11-59. The modified host cell of any one of Embodiments II- 47 to 11-58, wherein the modified host cell comprises two or more heterologous nucleic acids comprising a nucleotide sequence encoding KAR2 polypeptide.

[001482] Embodiment 11-60. The modified host cell of any one of Embodiments II- 47 to 11-58, wherein the modified host cell comprises three or more heterologous nucleic acids comprising a nucleotide sequence encoding a KAR2 polypeptide.

[001483] Embodiment 11-61. The modified host cell of Embodiment 11-47, wherein the one or more chaperone or co-chaperone polypeptides comprise a SSB1 polypeptide. [001484] Embodiment 11-62. The modified host cell of any one of Embodiments II- 45 to 11-61, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response.

[001485] Embodiment 11-63. The modified host cell of Embodiment 11-62, wherein the one or more polypeptides involved in unfolded protein response comprise an IRE1 polypeptide, a HACls polypeptide, a DED1 polypeptide, or a PPQ1 polypeptide, or a combination of any of the foregoing.

[001486] Embodiment 11-64. The modified host cell of Embodiment 11-63, wherein the one or more polypeptides involved in unfolded protein response comprise an IRE1 polypeptide.

[001487] Embodiment 11-65. The modified host cell of Embodiment 11-63 or Embodiment 11-64, wherein the one or more polypeptides involved in unfolded protein response comprise a HACls polypeptide.

[001488] Embodiment 11-66. The modified host cell of any one of Embodiments II- 63 to 11-65, wherein the one or more polypeptides involved in unfolded protein response comprise a PPQ1 polypeptide.

[001489] Embodiment 11-67. The modified host cell of Embodiment 11-62, wherein the one or more polypeptides involved in unfolded protein response comprise a transcription factor polypeptide or a lumenal sensor polypeptide that binds to an unfolded protein response element.

[001490] Embodiment 11-68. The modified host cell of any one of Embodiments II- 45 to 11-67, wherein the modified host cell comprises a synthetic polypeptide that binds to an unfolded protein response element.

[001491] Embodiment 11-69. The modified host cell of any one of Embodiments II- 45 to 11-68, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum.

[001492] Embodiment 11-70. The modified host cell of Embodiment 11-69, wherein the one or more polypeptides involved in protein translocation into the endoplasmic reticulum comprise a KAR2 polypeptide, a SEC61 polypeptide, a SBH1 polypeptide, a SSS1 polypeptide, a SRP14 polypeptide, a SRP21 polypeptide, a SRP68 polypeptide, a SRP72 polypeptide, a SEC65 polypeptide, a SRP54 polypeptide, a SEC62 polypeptide, a SEC63 polypeptide, a SEC66 polypeptide, a SEC72 polypeptide, a SRP101 polypeptide, or a SRP102 polypeptide, or a combination of any of the foregoing.

[001493] Embodiment 11-71. The modified host cell of Embodiment 11-70, wherein the one or more polypeptides involved in protein translocation into the endoplasmic reticulum comprise a SEC61 polypeptide, a SBH1 polypeptide, a SSS1 polypeptide, a SRP14 polypeptide, a SRP21 polypeptide, a SRP68 polypeptide, a SRP72 polypeptide, a SEC65 polypeptide, and a SRP54 polypeptide.

[001494] Embodiment 11-72. The modified host cell of Embodiment 11-70, wherein the one or more polypeptides involved in protein translocation into the endoplasmic reticulum comprise a SRP14 polypeptide, a SRP21 polypeptide, a SRP68 polypeptide, a SRP72 polypeptide, a SEC65 polypeptide, and a SRP54 polypeptide.

[001495] Embodiment 11-73. The modified host cell of any one of Embodiments II- 69 to 11-72, wherein the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding SRP protein complex RNA component SCR1.

[001496] Embodiment 11-74. The modified host cell of any one of Embodiments II- 45 to 11-73, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides

[001497] Embodiment 11-75. The modified host cell of Embodiment 11-74, wherein the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide.

[001498] Embodiment 11-76. The modified host cell of any one of Embodiments II- 45 to 11-75, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin mononucleotide (FMN) synthetase polypeptides.

[001499] Embodiment 11-77. The modified host cell of Embodiment 11-76, wherein the one or more flavin mononucleotide (FMN) synthetase polypeptides comprise a FMN1 polypeptide.

[001500] Embodiment 11-78. The modified host cell of any one of Embodiments II- 45 to 11-77, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosidase polypeptides.

[001501] Embodiment 11-79. The modified host cell of Embodiment 11-78, wherein the one or more glycosidase polypeptides comprise a CWH41 polypeptide.

[001502] Embodiment 11-80. The modified host cell of any one of Embodiments II- 45 to 11-79, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more fatty acid desaturase polypeptides.

[001503] Embodiment 11-81. The modified host cell of Embodiment 11-80, wherein the one or more fatty acid desaturase polypeptides comprise an OLE1 polypeptide.

[001504] Embodiment 11-82. The modified host cell of any one of Embodiments II- 45 to 11-81, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides.

[001505] Embodiment 11-83. The modified host cell of Embodiment 11-82, wherein the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide.

[001506] Embodiment 11-84. The modified host cell of any one of Embodiments II- 45 to 11-83, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides.

[001507] Embodiment 11-85. The modified host cell of Embodiment 11-84, wherein the one or more thiol oxidase polypeptides comprise an EROl polypeptide or an ERV2 polypeptide, or a combination of any of the foregoing.

[001508] Embodiment 11-86. The modified host cell of Embodiment 11-85, wherein the one or more thiol oxidase polypeptides comprise an EROl polypeptide.

[001509] Embodiment 11-87. The modified host cell of Embodiment 11-85 or Embodiment 11-86, wherein the modified host cell comprises two or more heterologous nucleic acids comprising a nucleotide sequence encoding an EROl polypeptide.

[001510] Embodiment 11-88. The modified host cell of any one of Embodiments II- 85 to 11-87, wherein the one or more thiol oxidase polypeptides comprise an ERV2 polypeptide. [001511] Embodiment 11-89. The modified host cell of any one of Embodiments II- 45 to 11-88, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more signal peptidase or signal peptidase complex polypeptides.

[001512] Embodiment 11-90. The modified host cell of Embodiment 11-89, wherein the one or more signal peptidase or signal peptidase complex polypeptides comprise a KEX2 polypeptide, a SPC1 polypeptide, a SPC2 polypeptide, a SPC3 polypeptide, or an SEC 11 polypeptide, or a combination of any of the foregoing.

[001513] Embodiment 11-91. The modified host cell of any one of Embodiments II- 45 to 11-90, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein transport and trafficking polypeptides.

[001514] Embodiment 11-92. The modified host cell of Embodiment 11-91, wherein the one or more protein transport and trafficking polypeptides comprise a SEC23

polypeptide, a SEC24 polypeptide, a SEC 13 polypeptide, a BFR2 polypeptide, a SEC31 polypeptide, a SAR1 polypeptide, a SEC 12 polypeptide, a SEC4 polypeptide, a SEC 16 polypeptide, an ERV29 polypeptide, a SEC7 polypeptide, a SEC5 polypeptide, a SEC22 polypeptide, a BOSl polypeptide, a BETl polypeptide, a BUGl polypeptide, a GRH1 polypeptide, a USOl polypeptide, a SEC17 polypeptide, a SEC18 polypeptide, a SSOl polypeptide, a SEC9 polypeptide, a SNC2 polypeptide, or a LDB17 polypeptide, or a combination of any of the foregoing.

[001515] Embodiment 11-93. The modified host cell of Embodiment 11-92, wherein the one or more protein transport and trafficking polypeptides comprise a BFR2 polypeptide.

[001516] Embodiment 11-94. The modified host cell of Embodiment 11-92 or Embodiment 11-93, wherein the one or more protein transport and trafficking polypeptides comprise a SEC24 polypeptide.

[001517] Embodiment 11-95. The modified host cell of any one of Embodiments II- 92 to 11-94, wherein the one or more protein transport and trafficking polypeptides comprise a LDB17 polypeptide.

[001518] Embodiment 11-96. The modified host cell of any one of Embodiments II- 45 to 11-95, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in endoplasmic reticulum-associated degradation (ERAD).

[001519] Embodiment 11-97. The modified host cell of Embodiment 11-96, wherein the one or more polypeptides involved in endoplasmic reticulum-associated degradation (ERAD) comprise a NPL4 polypeptide.

[001520] Embodiment 11-98. The modified host cell of any one of Embodiments II- 45 to 11-97, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cell wall assembly.

[001521] Embodiment 11-99. The modified host cell of Embodiment 11-98, wherein the one or more polypeptides involved in cell wall assembly comprise a KRE1 polypeptide.

[001522] Embodiment II- 100. The modified host cell of any one of Embodiments II- 45 to 11-99, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in vacuolar protein sorting.

[001523] Embodiment II- 101. The modified host cell of Embodiment II- 100, wherein the one or more polypeptides involved in vacuolar protein sorting comprise a PEP1 polypeptide.

[001524] Embodiment 11-102. The modified host cell of any one of Embodiments II- 45 to II- 101, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosyltransferase polypeptides.

[001525] Embodiment 11-103. The modified host cell of any one of Embodiments II- 43 to 11-102, wherein the modified host cell comprises a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides.

[001526] Embodiment 11-104. The modified host cell of Embodiment 11-103, wherein the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more glycosidase polypeptides, one or more genes encoding one or more glycosyltransferase polypeptides, one or more genes encoding one or more polypeptides involved in lipid droplet assembly, one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism, or one or more genes encoding one or more vacuolar proteinase polypeptides, or a combination of any of the foregoing.

[001527] Embodiment 11-105. The modified host cell of Embodiment 11-104, wherein the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more glycosidase polypeptides.

[001528] Embodiment 11-106. The modified host cell of Embodiment 11-105, wherein the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene or a MNS1 gene, or a combination of any of the foregoing.

[001529] Embodiment 11-107. The modified host cell of Embodiment 11-106, wherein the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene.

[001530] Embodiment 11-108. The modified host cell of any one of Embodiments II- 104 to 11-107, wherein the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more glycosyltransferase polypeptides.

[001531] Embodiment 11-109. The modified host cell of Embodiment 11-108, wherein the one or more genes encoding one or more glycosyltransferase polypeptides comprise an ALG12 gene.

[001532] Embodiment II- 110. The modified host cell of any one of Embodiments II- 104 to 11-109, wherein the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more polypeptides involved in lipid droplet assembly.

[001533] Embodiment II- 111. The modified host cell of Embodiment II- 110, wherein the one or more genes encoding one or more polypeptides involved in lipid droplet assembly comprise a FLD1 gene.

[001534] Embodiment II- 112. The modified host cell of any one of Embodiments II- 104 to II- 111, wherein the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism.

[001535] Embodiment II- 113. The modified host cell of Embodiment II-l 12, wherein the one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism comprise an OPI1 gene.

[001536] Embodiment II- 114. The modified host cell of any one of Embodiments II- 104 to II- 113, wherein the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides.

[001537] Embodiment II- 115. The modified host cell of Embodiment II-114, wherein the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene, a PRB1 gene, or a PRC1 gene, or a combination of any of the foregoing.

[001538] Embodiment II- 116. The modified host cell of Embodiment II-115, wherein the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene.

[001539] Embodiment II- 117. The modified host cell of any one of Embodiments II- 43 to II- 116, wherein the modified host cell comprises two or more heterologous nucleic acids comprising a nucleotide sequence encoding the cannabinoid synthase polypeptide.

[001540] Embodiment II- 118. The modified host cell of any one of Embodiments II- 43 to II- 117, wherein the nucleotide sequence encoding the cannabinoid synthase polypeptide is codon-optimized.

[001541] Embodiment II- 119. The modified host cell of any one of Embodiments II- 43 to II- 118, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide.

[001542] Embodiment 11-120. The modified host cell of any one of Embodiments II- 43 to II- 118, wherein the cannabinoid synthase polypeptide is a cannabichromenic acid synthase polypeptide.

[001543] Embodiment 11-121. The modified host cell of any one of Embodiments II- 43 to II- 118, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

[001544] Embodiment 11-122. The modified host cell of Embodiment 11-121, wherein the cannabidiolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 80% sequence identity to SEQ ID NO:242 or SEQ ID NO:244.

[001545] Embodiment 11-123. The modified host cell of Embodiment 11-121, wherein the cannabidiolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 85% sequence identity to SEQ ID NO:242 or SEQ ID NO:244.

[001546] Embodiment 11-124. The modified host cell of Embodiment 11-121, wherein the cannabidiolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 90% sequence identity to SEQ ID NO:242 or SEQ ID NO:244. [001547] Embodiment 11-125. The modified host cell of Embodiment 11-121, wherein the cannabidiolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 95% sequence identity to SEQ ID NO:242 or SEQ ID NO:244.

[001548] Embodiment 11-126. The modified host cell of Embodiment 11-121, wherein the cannabidiolic acid synthase polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:243 or SEQ ID NO:245.

[001549] Embodiment 11-127. The modified host cell of any one of Embodiments II- 43 to 11-126, wherein the cannabinoid synthase polypeptide comprises a signal sequence polypeptide.

[001550] Embodiment 11-128. The modified host cell of Embodiment 11-127, wherein the signal sequence polypeptide is a secretory signal sequence polypeptide.

[001551] Embodiment 11-129. The modified host cell of Embodiment 11-128, wherein the secretory signal sequence polypeptide is a native secretory signal sequence polypeptide.

[001552] Embodiment 11-130. The modified host cell of Embodiment 11-128, wherein the secretory signal sequence polypeptide is a synthetic secretory signal sequence polypeptide.

[001553] Embodiment 11-131. The modified host cell of Embodiment 11-128, wherein the secretory signal sequence polypeptide is an endoplasmic reticulum retention signal sequence polypeptide.

[001554] Embodiment 11-132. The modified host cell of Embodiment 11-128, wherein the secretory signal sequence polypeptide is a vacuolar localization signal sequence polypeptide.

[001555] Embodiment 11-133. The modified host cell of Embodiment 11-132, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide or a PRC It polypeptide.

[001556] Embodiment 11-134. The modified host cell of Embodiment 11-133, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide.

[001557] Embodiment 11-135. The modified host cell of Embodiment 11-128, wherein the secretory signal sequence polypeptide is a plasma membrane localization signal sequence polypeptide.

[001558] Embodiment 11-136. The modified host cell of Embodiment 11-128, wherein the secretory signal sequence polypeptide is a peroxisome targeting signal sequence polypeptide. [001559] Embodiment 11-137. The modified host cell of Embodiment 11-121, wherein the cannabidiolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 80% sequence identity to SEQ ID NO: l, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8.

[001560] Embodiment 11-138. The modified host cell of Embodiment 11-121, wherein the cannabidiolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 85% sequence identity to SEQ ID NO: l, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8.

[001561] Embodiment 11-139. The modified host cell of Embodiment 11-121, wherein the cannabidiolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 90% sequence identity to SEQ ID NO: l, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8.

[001562] Embodiment 11-140. The modified host cell of Embodiment 11-121, wherein the cannabidiolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 95% sequence identity to SEQ ID NO: l, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8.

[001563] Embodiment 11-141. The modified host cell of Embodiment 11-121, wherein the cannabidiolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 80% sequence identity to SEQ ID NO:256, SEQ ID NO:258, SEQ ID N0 259, SEQ ID NO 260, SEQ ID N0 261, SEQ ID N0 262, or SEQ ID N0 263

[001564] Embodiment 11-142. The modified host cell of Embodiment 11-121, wherein the cannabidiolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 85% sequence identity to SEQ ID NO:256, SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:261, SEQ ID NO:262, or SEQ ID NO:263.

[001565] Embodiment 11-143. The modified host cell of Embodiment 11-121, wherein the cannabidiolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 90% sequence identity to SEQ ID NO:256, SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO:263.

[001566] Embodiment 11-144. The modified host cell of Embodiment 11-121, wherein the cannabidiolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 95% sequence identity to SEQ ID NO:256, SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO:263.

[001567] Embodiment II-144a. The modified host cell of Embodiment 11-121, wherein the cannabidiolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence set forth in SEQ ID NO:389, SEQ ID NO:39l, SEQ ID NO:393, SEQ ID NO:395, SEQ ID NO:397, SEQ ID NO:399, SEQ ID NO:40l, SEQ ID NO:403, SEQ ID NO:405, SEQ ID NO:407, SEQIDNO:409, SEQIDNO:41l, SEQIDNO:4l3, SEQIDNO:4l5, SEQ ID NO:417, SEQ ID NO:419, SEQ ID NO:42l, SEQ ID NO:423, SEQ ID NO:425, SEQ ID NO:427, SEQIDNO:429, SEQIDNO:43l, SEQIDNO:433, SEQIDNO:435, SEQ ID NO:437, SEQ ID NO:439, SEQ ID NO:44l, SEQ ID NO:443, SEQ ID NO:445, SEQ ID NO:447, SEQIDNO:449, SEQIDNO:45l, SEQIDNO:453, SEQIDNO:455, SEQ ID NO:457, SEQ ID NO:459, SEQ ID NO:46l, SEQ ID NO:463, SEQ ID NO:465, SEQ ID NO:467, SEQ ID NO:469, SEQ ID NO:47l, SEQ ID NO:473, SEQ ID NO:475, SEQ ID NO:477, SEQIDNO:479, SEQIDNO:48l, SEQIDNO:483, SEQIDNO:485, SEQ ID NO:487, SEQ ID NO:489, SEQ ID NO:49l, SEQ ID NO:493, SEQ ID NO:495, SEQ ID NO:497, SEQ ID NO:499, SEQ ID NO:50l, SEQ ID NO:503, SEQ ID NO:505, SEQ ID NO:507, SEQIDNO:509, SEQIDNO:51l, SEQIDNO:5l3, SEQIDNO:5l5, SEQ ID NO:517, SEQIDNO:519, SEQIDNO:52l, SEQIDNO:523, SEQIDN0525, SEQ ID NO: 527, SEQIDNO:529, SEQIDNO:53l, SEQIDNO:533, SEQIDNO:535, SEQ ID NO:537, SEQ ID NO:539, SEQ ID NO:54l, SEQ ID NO:543, SEQ ID NO:545, SEQ ID NO: 547, SEQIDNO:549, SEQIDNO:55l, SEQIDNO:553, SEQIDN0555, SEQ ID NO:557, SEQ ID NO:559, SEQ ID NO:56l, SEQ ID NO:563, SEQ ID NO:565, SEQ ID NO:567, SEQ ID NO:569, SEQ ID NO:57l, SEQ ID NO:573, SEQ ID NO:579, SEQIDN0581, SEQIDN0583, SEQIDNO:585, SEQIDN0587, SEQIDN0589, SEQ ID NO:59l, SEQ ID NO:593, SEQ ID NO:595, SEQ ID NO:597, or SEQ ID NO:599.

[001568] Embodiment 11-145. The modified host cell of Embodiment II- 119, wherein the tetrahydrocannabinolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 80% sequence identity to SEQ ID NO:238, SEQ ID NO:27l, SEQ ID NO:272, SEQ ID NO:273, or SEQ ID NO:274.

[001569] Embodiment 11-146. The modified host cell of Embodiment II- 119, wherein the tetrahydrocannabinolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 85% sequence identity to SEQ ID NO:238, SEQ ID NO:27l, SEQ ID NO:272, SEQ ID N0273, or SEQ ID NO:274.

[001570] Embodiment 11-147. The modified host cell of Embodiment II- 119, wherein the tetrahydrocannabinolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 90% sequence identity to SEQ ID NO:238, SEQ ID NO:27l, SEQ ID NO:272, SEQ ID NO:273, or SEQ ID NO:274. [001571] Embodiment 11-148. The modified host cell of Embodiment II-l 19, wherein the tetrahydrocannabinolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 95% sequence identity to SEQ ID NO:238, SEQ ID NO:27l, SEQ ID NO:272, SEQ ID NO:273, or SEQ ID NO:274.

[001572] Embodiment 11-149. The modified host cell of Embodiment II- 119, wherein the tetrahydrocannabinolic acid synthase polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:239.

[001573] Embodiment 11-150. The modified host cell of Embodiment 11-120, wherein the cannabichromenic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 80% sequence identity to SEQ ID NO:240.

[001574] Embodiment 11-151. The modified host cell of Embodiment 11-120, wherein the cannabichromenic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 80% sequence identity to SEQ ID NO:240.

[001575] Embodiment 11-152. The modified host cell of Embodiment 11-120, wherein the cannabichromenic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 90% sequence identity to SEQ ID NO:240.

[001576] Embodiment 11-153. The modified host cell of Embodiment 11-120, wherein the cannabichromenic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 95% sequence identity to SEQ ID NO:240.

[001577] Embodiment 11-154. The modified host cell of Embodiment 11-120, wherein the cannabichromenic acid synthase polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:241.

[001578] Embodiment 11-155. The modified host cell of Embodiment 11-121, wherein the cannabidiolic acid synthase polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:9.

[001579] Embodiment 11-156. The modified host cell of Embodiment 11-121, wherein the cannabidiolic acid synthase polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:257.

[001580] Embodiment II-156a. The modified host cell of Embodiment 11-121, wherein the cannabidiolic acid synthase polypeptide is a variant cannabidiolic acid synthase polypeptide comprising an amino acid sequence set forth in SEQ ID NO:390, SEQ ID NO:392, SEQ ID NO:394, SEQ ID NO:396, SEQ ID N0 398, SEQ ID NO:400, SEQ ID NO:402, SEQ ID NO:404, SEQ ID NO:406, SEQ ID NO:408, SEQ ID NO:410, SEQ ID NO:412, SEQ ID NO:414, SEQ ID NO:416, SEQ ID NO:418, SEQ ID NO:420, SEQ ID NO:422, SEQ ID NO:424, SEQ ID NO:426, SEQ ID N0 428, SEQ ID NO:430, SEQ ID NO:432, SEQ ID NO:434, SEQ ID NO:436, SEQ ID N0 438, SEQ ID NO:440, SEQ ID NO:442, SEQ ID NO:444, SEQ ID NO:446, SEQ ID NO:448, SEQ ID NO:450, SEQ ID NO:452, SEQ ID NO:454, SEQ ID NO:456, SEQ ID N0 458, SEQ ID NO:460, SEQ ID NO:462, SEQ ID NO:464, SEQ ID NO:466, SEQ ID N0 468, SEQ ID NO:470, SEQ ID NO:472, SEQ ID NO:474, SEQ ID NO:476, SEQ ID NO:478, SEQ ID NO:480, SEQ ID NO:482, SEQ ID NO:484, SEQ ID NO:486, SEQ ID NO:488, SEQ ID NO:490, SEQ ID NO:492, SEQ ID NO:494, SEQ ID NO:496, SEQ ID NO:498, SEQ ID NO:500, SEQ ID NO:502, SEQ ID NO: 504, SEQ ID NO:506, SEQ ID NO 508, SEQ ID NO:510, SEQ ID NO:5l2, SEQ ID NO: 514, SEQ ID NO:5 l6, SEQ ID N0 518, SEQ ID NO:520, SEQ ID NO:522, SEQ ID NO:524, SEQ ID N0 526, SEQ ID N0 528, SEQ ID NO:530, SEQ ID NO:532, SEQ ID NO:534, SEQ ID NO:536, SEQ ID N0 538, SEQ ID NO:540, SEQ ID NO:542, SEQ ID NO:544, SEQ ID NO:546, SEQ ID N0 548, SEQ ID NO:550, SEQ ID NO:552, SEQ ID NO:554, SEQ ID N0 556, SEQ ID N0 558, SEQ ID NO:560, SEQ ID NO:562, SEQ ID NO:564, SEQ ID NO:566, SEQ ID N0 568, SEQ ID NO:570, SEQ ID NO: 572, SEQ ID NO: 574, SEQ ID NO: 580, SEQ ID NO: 582, SEQ ID NO: 584, SEQ ID NO:586, SEQ ID NO:588, SEQ ID NO 590, SEQ ID N0 592, SEQ ID NO:594, SEQ ID NO:596, SEQ ID NO:598, or SEQ ID NO:600.

[001581] Embodiment 11-157. The modified host cell of any one of Embodiments II- 43 to II- 156a, wherein the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate: olivetolic acid geranyl transferase (GOT) polypeptide.

[001582] Embodiment 11-158. The modified host cell of Embodiment 11-157, wherein the GOT polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:70 or SEQ ID NO:73.

[001583] Embodiment II-l 58a The modified host cell of Embodiment 11-157 or II-

158, wherein the modified host cell comprises two or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide.

[001584] Embodiment II- 158b. The modified host cell of any one of Embodiments II- 43 to 11-156, wherein the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a NphB polypeptide.

[001585] Embodiment II- 158c. The modified host cell of Embodiment II- 158b, wherein the NphB polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:576. [001586] Embodiment 11-159. The modified host cell of any one of Embodiments II- 43 to II- 158c, wherein the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a tetraketide synthase (TKS) polypeptide and one or more heterologous nucleic acids comprising a nucleotide sequence encoding an olivetolic acid cyclase (OAC) polypeptide.

[001587] Embodiment 11-160. The modified host cell of Embodiment 11-159, wherein the TKS polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:75.

[001588] Embodiment 11-161. The modified host cell of Embodiment 11-159 or Embodiment 11-160, wherein the modified host cell comprises three or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide.

[001589] Embodiment 11-162. The modified host cell of any one of Embodiments II- 159 to 11-161, wherein the OAC polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:77 or SEQ ID NO:225.

[001590] Embodiment 11-163. The modified host cell of any one of Embodiments II- 159 to 11-162, wherein the modified host cell comprises three or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide.

[001591] Embodiment 11-164. The modified host cell of any one of Embodiments II- 43 to 11-163, wherein the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acyl-activating enzyme (AAE) polypeptide.

[001592] Embodiment 11-165. The modified host cell of Embodiment 11-164, wherein the AAE polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:79, SEQ ID NO:268, or SEQ ID NO:270.

[001593] Embodiment 11-166. The modified host cell of Embodiment 11-164 or Embodiment 11-165, wherein the modified host cell comprises two or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide.

[001594] Embodiment 11-167. The modified host cell of any one of Embodiments II- 43 to 11-166, wherein the modified host cell comprises one or more of the following: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMG-CoA synthase (HMGS) polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3-hydroxy-3-methyl-glutaryl-CoA reductase (tHMGR) polypeptide; c) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate kinase (MK) polypeptide; d) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a phosphomevalonate kinase (PMK) polypeptide; e) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate pyrophosphate decarboxylase (MVD1) polypeptide; or f) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a isopentenyl diphosphate isomerase (IDI1) polypeptide.

[001595] Embodiment 11-168. The modified host cell of Embodiment 11-167, wherein the IDI1 polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 81.

[001596] Embodiment 11-169. The modified host cell of Embodiment 11-167 or Embodiment 11-168, wherein the tHMGR polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:83.

[001597] Embodiment 11-170. The modified host cell of any one of Embodiments II- 167 to 11-169, wherein the HMGS polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:85.

[001598] Embodiment 11-171. The modified host cell of any one of Embodiments II- 167 to 11-170, wherein the MK polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:95.

[001599] Embodiment 11-172. The modified host cell of any one of Embodiments II- 167 to 11-171, wherein the PMK polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:93

[001600] Embodiment 11-173. The modified host cell of any one of Embodiments II- 167 to 11-172, wherein the MVD1 polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:89

[001601] Embodiment 11-174. The modified host cell of any one of Embodiments II- 43 to 11-173, wherein the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide.

[001602] Embodiment 11-175. The modified host cell of Embodiment 11-174, wherein the acetoacetyl-CoA thiolase polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:87.

[001603] Embodiment 11-176. The modified host cell of any one of Embodiments II- 43 to 11-175, wherein the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a pyruvate decarboxylase (PDC) polypeptide. [001604] Embodiment 11-177. The modified host cell of Embodiment 11-176, wherein the PDC polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:91.

[001605] Embodiment 11-178. The modified host cell of any one of Embodiments II- 43 to 11-177, wherein the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate synthetase (GPPS) polypeptide.

[001606] Embodiment 11-179. The modified host cell of Embodiment 11-178, wherein the GPPS polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:97.

[001607] Embodiment 11-180. The modified host cell of any one of Embodiments II- 43 to 11-179, wherein the modified host cell is a eukaryotic cell.

[001608] Embodiment 11-181. The modified host cell of Embodiment 11-180, wherein the eukaryotic cell is a yeast cell.

[001609] Embodiment 11-182. The modified host cell of Embodiment 11-181, wherein the yeast cell is Saccharomyces cerevisiae.

[001610] Embodiment 11-183. The modified host cell of Embodiment 11-182, wherein the Saccharomyces cerevisiae is a protease-deficient strain of Saccharomyces cerevisiae.

[001611] Embodiment 11-184. The modified host cell of any one of Embodiments II- 43 to 11-183, wherein at least one of the one or more heterologous nucleic acids are integrated into the chromosome of the modified host cell.

[001612] Embodiment 11-185. The modified host cell of any one of Embodiments II- 43 to 11-183, wherein at least one of the one or more heterologous nucleic acids are maintained extrachromosomally.

[001613] Embodiment 11-186. The modified host cell of any one of Embodiments II- 43 to 11-185, wherein at least one of the one or more heterologous nucleic acids are operably- linked to an inducible promoter.

[001614] Embodiment 11-187. The modified host cell of any one of Embodiments II- 43 to 11-185, wherein at least one of the one or more heterologous nucleic acids are operably- linked to a constitutive promoter.

[001615] Embodiment 11-188. A method of producing a cannabinoid or a cannabinoid derivative, the method comprising culturing a modified host cell of any one of Embodiments 11-34 to 11-187 in a culture medium. [001616] Embodiment II- 188a. The method of Embodiment 11-188, wherein the method comprises recovering the produced cannabinoid or cannabinoid derivative.

[001617] Embodiment 11-189. The method of Embodiment 11-188 or II-188a, wherein the culture medium comprises a carboxylic acid.

[001618] Embodiment 11-190. The method of Embodiment 11-189, wherein the carboxylic acid is an unsubstituted or substituted C3-C18 carboxylic acid.

[001619] Embodiment 11-191. The method of Embodiment 11-190, wherein the unsubstituted or substituted C3-C 18 carboxylic acid is an unsubstituted or substituted hexanoic acid.

[001620] Embodiment 11-192. The method of Embodiment 11-188 or II-l88a, wherein the culture medium comprises olivetolic acid or an olivetolic acid derivative.

[001621] Embodiment 11-193. The method of any one of Embodiments 11-188 to II- 192, wherein the culture medium comprises a fermentable sugar.

[001622] Embodiment 11-194. The method of any one of Embodiments 11-188 to II- 192, wherein the culture medium comprises a pretreated cellulosic feedstock.

[001623] Embodiment 11-195. The method of any one of Embodiments 11-188 to II- 192, wherein the culture medium comprises a non-fermentable carbon source.

[001624] Embodiment 11-196. The method of Embodiment 11-195, wherein the non- fermentable carbon source comprises ethanol.

[001625] Embodiment 11-197. The method of any one of Embodiments 11-188 to II-

196, wherein the cannabinoid is cannabigerolic acid, cannabigerol, D⁹- tetrahydrocannabinolic acid, D⁹ -tetrahydrocannabinol, A⁸-tetrahydrocannabinolic acid, D⁸- tetrahydrocannabinol, cannabidiolic acid, cannabidiol, cannabichromenic acid,

cannabichromene, cannabinolic acid, cannabinol, cannabidivarinic acid, cannabidivarin, tetrahydrocannabivarinic acid, tetrahydrocannabivarin, cannabichromevarinic acid, cannabichromevarin, cannabigerovarinic acid, cannabigerovarin, cannabicyclolic acid, cannabicyclol, cannabielsoinic acid, cannabielsoin, cannabicitranic acid, or cannabicitran.

[001626] Embodiment 11-198. The method of any one of Embodiments 11-188 to II-

197, wherein the cannabinoid or cannabinoid derivative is produced in an amount of more than 100 mg/L culture medium.

[001627] Embodiment 11-199. A method of making a modified host cell of any one of Embodiments 11-43 to 11-187, the method comprising introducing into a host cell: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and b) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides.

[001628] Embodiment 11-200. A method of making a modified host cell of any one of Embodiments 11-43 to 11-187, the method comprising introducing into a host cell: a) one or more vectors comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and b) one or more vectors comprising one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides.

[001629] Embodiment 11-201. The method of Embodiment 11-199 or Embodiment II- 200, the method comprising introducing into the host cell a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides.

[001630] Some embodiments of the disclosure are of Embodiment III:

[001631] Embodiment III- 1. A compound of Formula (I):

or a pharmaceutically acceptable salt thereof, wherein,

[001632] Embodiment III-2. The compound of Embodiment III- 1 , or a pharmaceutically acceptable salt thereof, wherein, R¹ is a Ci-Cisalkyl group substituted with R^a or R¹ is a C2-Cisalkenyl group substituted with R^a, wherein R^a is -S-Ci-C6alkyl or -S-C2-C6alkenyl, or R¹ is selected from the group consisting of:

[001633] Embodiment III-3. The compound of Embodiment III- 1 , or a pharmaceutically acceptable salt thereof, wherein,

R¹ is selected from the group consisting of:

[001634] Embodiment III-4. The compound of Embodiment III- 1 , or a pharmaceutically acceptable salt thereof, wherein,

R¹ is a Ci-Cisalkyl group substituted with R^a or R¹ is a C2-Cisalkenyl group substituted with R^a, wherein R^a is -S-Ci-C6alkyl or -S-C2-C6alkenyl.

[001635] Embodiment III-5. The compound of Embodiment III- 1 , or a pharmaceutically acceptable salt thereof, wherein, R¹ is selected from the group consisting of:

[001636] Embodiment III-6. The compound of Embodiment III- 1 , or a pharmaceutically acceptable salt thereof, wherein the compound of Formula (I) is selected from the group consisting of:

561

562

[001637] Embodiment III-7. The compound of Embodiment III- 1 , or a pharmaceutically acceptable salt thereof, wherein the compound of Formula (I) is selected from the group consisting of:

[001638] Embodiment III-8. The compound of Embodiment III- 1 , or a pharmaceutically acceptable salt thereof, wherein the compound of Formula (I) is selected from the group consisting of:

[001639] Embodiment III-9. A pharmaceutical composition comprising a compound of any one of Embodiments III-1 to III-8, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[001640] Embodiment III- 10. A method for treating a disorder in a subject, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of any one of Embodiments III-1 to III-8, or a pharmaceutically acceptable salt thereof.

[001641] Embodiment III- 11. A method for treating a disorder in a subject, comprising administering to a subject in need thereof a therapeutically effective amount of a pharmaceutical composition according to Embodiment III-9.

[001642] Embodiment III- 12. The method of Embodiment III- 10 or III-11, wherein the disorder is mediated by cannabinoid receptor type 1 or cannabinoid receptor type 2.

[001643] Embodiment III- 13 The method of Embodiment III- 10 or III- 11, wherein the disorder is chronic pain, multiple sclerosis, cancer-associated nausea and vomiting, weight loss, appetite loss, spasticity, or seizures.

[001644] Embodiment III- 1 . Use of a compound of any one of Embodiments III-1 to

III-8, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating a disorder in a subject in need thereof.

[001645] Embodiment III- 15. Use of a pharmaceutical composition according to Embodiment III-9 in the manufacture of a medicament for treating a disorder in a subject in need thereof. [001646] Embodiment III- 16. Use of a compound of any one of Embodiments III-1 to III-8, or a pharmaceutically acceptable salt thereof, for treating a disorder in a subject in need thereof.

[001647] Embodiment III- 17. Use of a pharmaceutical composition according to Embodiment III-9 for treating a disorder in a subject in need thereof.

[001648] Embodiment III- 18. The use of any one of Embodiments III- 14 to III-17, wherein the disorder is mediated by cannabinoid receptor type 1 or cannabinoid receptor type 2.

[001649] Embodiment III- 19. The use of any one of Embodiments III- 14 to III-17, wherein the disorder is chronic pain, multiple sclerosis, cancer-associated nausea and vomiting, weight loss, appetite loss, spasticity, or seizures.

[001650] Embodiment III-20. A compound of any one of Embodiments III- 1 to III-8, or a pharmaceutically acceptable salt thereof, for use in a method of treating a disorder in a subj ect in need thereof.

[001651] Embodiment III-21. The compound for use of Embodiment III-20, wherein the disorder is mediated by cannabinoid receptor type 1 or cannabinoid receptor type 2.

[001652] Embodiment III-22. The compound for use of Embodiment III-20, wherein the disorder is chronic pain, multiple sclerosis, cancer-associated nausea and vomiting, weight loss, appetite loss, spasticity, or seizures.

[001653] Embodiment III-23 A pharmaceutical composition according to

Embodiment III-9 for use in a method of treating a disorder in a subject in need thereof.

[001654] Embodiment III-24. The pharmaceutical composition for use of

Embodiment III-23, wherein the disorder is mediated by cannabinoid receptor type 1 or cannabinoid receptor type 2.

[001655] Embodiment III-25. The pharmaceutical composition for use of

Embodiment III-23, wherein the disorder is chronic pain, multiple sclerosis, cancer- associated nausea and vomiting, weight loss, appetite loss, spasticity, or seizures.

[001656] Embodiment III-26. A composition comprising a compound of any one of Embodiments III- 1 to III-8, or a pharmaceutically acceptable salt thereof, and an acceptable carrier.

[001657] Some embodiments of the disclosure are of Embodiment IV:

[001658] Embodiment IV- 1. A method of producing a cannabinoid or a cannabinoid derivative comprising: a) culturing a modified host cell of any one of Embodiments 1-34 to 1-187 in a culture medium; and

b) recovering the produced cannabinoid or cannabinoid derivative.

[001659] Embodiment IV-2. The method of Embodiment IV- 1, wherein the culture medium comprises a carboxylic acid.

[001660] Embodiment IV-3. The method of Embodiment IV-2, wherein the carboxylic acid is an unsubstituted or substituted C3-C18 carboxylic acid.

[001661] Embodiment IV-4. The method of Embodiment IV-3, wherein the unsubstituted or substituted C3-C 18 carboxylic acid is an unsubstituted or substituted hexanoic acid.

[001662] Embodiment IV-5. The method of Embodiment IV- 1, wherein the culture medium comprises olivetolic acid or an olivetolic acid derivative.

[001663] Embodiment IV-6. The method of Embodiment IV-l, wherein the cannabinoid is cannabigerolic acid, cannabigerol, A⁹-tetrahydrocannabinolic acid, D⁹- tetrahydrocannabinol, A⁸-tetrahydrocannabinolic acid, A⁸-tetrahydrocannabinol,

cannabidiolic acid, cannabidiol, cannabichromenic acid, cannabichromene, cannabinolic acid, cannabinol, cannabidivarinic acid, cannabidivarin, tetrahydrocannabivarinic acid, tetrahydrocannabivarin, cannabichromevarinic acid, cannabichromevarin,

cannabigerovarinic acid, cannabigerovarin, cannabicyclolic acid, cannabicyclol,

cannabielsoinic acid, cannabielsoin, cannabicitranic acid, or cannabicitran.

[001664] Embodiment IV-7. The method of Embodiment IV-2, wherein the carboxylic acid is selected from the group consisting of butyric acid, valeric acid, hexanoic acid, octanoic acid, 2-methylhexanoic acid, 3-methylhexanoic acid, 4-methylhexanoic acid, 5-methylhexanoic acid, 2-hexenoic acid, 3-hexenoic acid, 4-hexenoic acid, 5-hexenoic acid, heptanoic acid, 5-chlorovaleric acid, 5-(methylsulfanyl)valeric acid, 4-pentynoic acid, trans- 2-pentenoic acid, 5-hexynoic acid, trans-2-hexenoic acid, 6-heptynoic acid, trans-2-octenoic acid, nonanoic acid, trans-2-nonenoic acid, decanoic acid, undecanoic acid, dodecanoic acid, myristic acid, 4-phenylbutyric acid, 5-phenylvaleric acid, 6-phenylhexanoic acid, 7- phenylheptanoic acid, isobutyric acid, fumaric acid, itaconic acid, malic acid, succinic acid, maleic acid, malonic acid, glutaric acid, glucaric acid, oxalic acid, adipic acid, pimelic acid, suberic acid, azelaic acid, sebacic acid, dodecandioic acid, glutaconic acid, ortho-phthalic acid, isophthalic acid, terephthalic acid, citric acid, isocitric acid, aconitic acid, tricarballylic acid, trimesic acid, 5-aminovaleric acid, 5-cyanovaleric acid, 5-hydroxyvaleric acid, and 2,3- dimethylhexanoic acid. [001665] Embodiment IV-8. The method of Embodiment IV-2, wherein the carboxylic acid is selected from the group consisting of 2-methylhexanoic acid, 3- methylhexanoic acid, 4-methylhexanoic acid, 5-methylhexanoic acid, 2-hexenoic acid, 3- hexenoic acid, 4-hexenoic acid, heptanoic acid, 5-chlorovaleric acid, 5- (methylsulfanyl)valeric acid, 4-pentynoic acid, trans-2-pentenoic acid, 5-hexynoic acid, trans-2-hexenoic acid, 6-heptynoic acid, trans-2-octenoic acid, nonanoic acid, trans-2- nonenoic acid, decanoic acid, undecanoic acid, dodecanoic acid, myristic acid, 4- phenylbutyric acid, 5-phenylvaleric acid, 6-phenylhexanoic acid, 7-phenylheptanoic acid, isobutyric acid, fumaric acid, itaconic acid, malic acid, maleic acid, glucaric acid, suberic acid, azelaic acid, sebacic acid, dodecandioic acid, glutaconic acid, ortho-phthalic acid, isophthalic acid, terephthalic acid, citric acid, isocitric acid, aconitic acid, tricarballylic acid, trimesic acid, 5-aminovaleric acid, 5-cyanovaleric acid, 5 -hydroxy valeric acid, and 2,3- dimethylhexanoic acid.

[001666] Embodiment IV-9. The method of Embodiment IV-2, wherein the carboxylic acid is selected from the group consisting of 4-pentynoic acid, trans-2-pentenoic acid, 5-hexynoic acid, trans-2-hexenoic acid, 6-heptynoic acid, trans-2-octenoic acid, nonanoic acid, trans-2-nonenoic acid, decanoic acid, undecanoic acid, dodecanoic acid, 4- phenylbutyric acid, 5-phenylvaleric acid, 6-phenylhexanoic acid, and 7-phenylheptanoic acid.

[001667] Embodiment IV-10. The method of Embodiment IV-2, wherein the carboxylic acid is selected from the group consisting of 2-methylhexanoic acid, 4- methylhexanoic acid, 5-methylhexanoic acid, 2-hexenoic acid, 3-hexenoic acid, 4-hexenoic acid, heptanoic acid, 5-chlorovaleric acid, and 5-(methylsulfanyl)valeric acid.

[001668] Embodiment IV-11. The method of Embodiment IV-2, wherein the carboxylic acid is a carboxylic acid of Formula (II):

wherein R is a Ci-C ixalkyl group substituted with R^a or a C2-C ixalkenyl group substituted with R^a, wherein R^a is -S-Ci-C6alkyl or -S-C2-C6alkenyl.

[001669] Embodiment IV- 12. The method of any one of Embodiments IV-1 to IV-11, wherein the culture medium comprises a fermentable sugar.

[001670] Embodiment IV-13. The method of any one of Embodiments IV-1 to IV-11, wherein the culture medium comprises a pretreated cellulosic feedstock. [001671] Embodiment IV-14. The method of any one of Embodiments IV-1 to IV-l 1, wherein the culture medium comprises a non-fermentable carbon source.

[001672] Embodiment IV-15. The method of Embodiment IV-14, wherein the non- fermentable carbon source comprises ethanol.

[001673] Embodiment IV-16. The method of any one of Embodiments IV-1 to IV-15, wherein the cannabinoid or cannabinoid derivative is produced in a recoverable amount of more than 100 mg/L culture medium.

[001674] Embodiment IV- 17. The method of any one of Embodiments IV-1 to IV-15, wherein the cannabinoid or cannabinoid derivative is produced in a recoverable amount of more than 50 mg/L culture medium.

[001675] Provided in Table 1 are amino acid and nucleotide sequences disclosed herein. Where a genus and/or species is noted, the sequence should not be construed to be limited only to the specified genus and/or species, but also includes other genera and/or species expressing said sequence. Orthologs of the sequences disclosed in Table 1 may also be encompassed by this disclosure. Nucleotide sequences indicated as codon optimized in Table 1 are codon optimized for expression in S. cerevisiae. In Table 1, used as the end of a sequence denotes a stop codon. In reference to OAC*, denotes a mutation is present in the sequence.

Table 1: Amino acid and nucleotide sequences of the disclosure

111

EXAMPLES

[001676] The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the present disclosure, and are not intended to limit the scope of what the inventors regard as their disclosure nor are they intended to represent that the experiments below are all or the only experiments performed. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperature, etc.) but some experimental errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, molecular weight is weight average molecular weight, temperature is in degrees Celsius, and pressure is at or near atmospheric. Standard abbreviations may be used, e.g., bp, base pair(s); kb, kilobase(s); s or sec, second(s); min, minute(s); h or hr, hour(s); aa, amino acid(s); bp, base pair(s); nt, nucleotide(s); and the like.

General Methods of the Examples

Yeast transformation methods

[001677] Each DNA construct comprising one or more heterologous nucleic acids disclosed herein (e.g., constructs detailed in Tables 2, 3, and 9) was integrated into

Saccharomyces cerevisiae (CEN.PK2, Strain S4) with standard molecular biology techniques in an optimized lithium acetate (LiAc) transformation. Briefly, cells were grown overnight in yeast extract peptone dextrose (YPD) media at 30 °C with shaking (200 rpm), diluted to an OD600 of 0.1 in 100 mL YPD, and grown to an OD600 of 0.6 - 0.8. For each transformation, 5 mL of culture was harvested by centrifugation, washed in 5 mL of sterile water, spun down again, resuspended in 1 mL of 100 mM LiAc, and transferred to a microcentrifuge tube. Cells were spun down (13,000 x g) for 30 seconds, the supernatant was removed, and the cells were resuspended in a transformation mix consisting of 240 pL 50% PEG, 36 pL 1M LiAc, 10 pL boiled salmon sperm DNA, and 74 pL of donor DNA. Following a heat shock at 42 °C for 40 minutes, cells were recovered overnight in YPD media before plating on selective media. DNA integration was confirmed by colony PCR with primers specific to the integrations.

Yeast culturing conditions

[001678] Yeast colonies verified to contain the expected DNA assembly comprising one or more heterologous nucleic acids disclosed herein, modified host cells, were picked into 96-well microtiter plates containing 360 pL of YPD (10 g/L yeast extract, 20 g/L Bacto peptone, 20 g/L dextrose (glucose)) and sealed with a breathable film seal. Cells were cultured at 30 °C in a high capacity microtiter plate incubator shaking at 1000 rpm and 80% humidity for 2 days until the cultures reached carbon exhaustion. The growth-saturated cultures were subcultured into fresh plates containing base media and either olivetolic acid or hexanoic acid, or an olivetolic acid derivative or a carboxylic acid other than hexanoic acid (either 1 mM olivetolic acid or 2 mM hexanoic acid, or 1 mM of an olivetolic acid derivative or 2 mM of a carboxylic acid other than hexanoic acid), by taking 14.4 pL from the saturated cultures and diluting into 360 uL of fresh media and sealed with a breathable film seal. The base media was either YPGal (10 g/L yeast extract, 20 g/L Bacto peptone, 20 or 40 g/L galactose, 1 g/L glucose) or defined minimal media (40 g/L galactose, 1 g/L glucose, 15 g/L ammonium sulfate, 8 g/L potassium phosphate, 6.15 g/L magnesium sulfate, 5.9 g/L succinate, 60 pg/L D-biotin, 120 pg/L Ca-D-pantothenate, 120 pg/L nicotinic acid, 300 pg/L myo-inositol, 60 pg/L thiamine hydrochloride, 60 pg/L pyridoxal hydrochloride, 2.5 pg/L p-aminobenzoic acid, 150 mg/L EDTA, 57.5 mg/L zinc sulfate heptahydrate, 2.6 mg/L manganese chloride dihydrate, 4.7 mg/L cobalt chloride hexahydrate, 5 mg/L copper sulfate pentahydrate, 4.8 mg/L sodium molybdate dihydrate, 29 mg/L calcium chloride dihydrate, and 28 mg/L iron sulfate heptahydrate). Modified host cells in the production media were cultured at 30 °C in a high capacity microtiter plate shaker at 1000 rpm and 80% humidity for an additional 3, 5, or 7 days prior to extraction and analysis. Upon completion, 100 pL of whole cell broth was diluted into 900 pL of methanol, sealed with a foil seal, and shaken at 1500 rpm for 60 seconds to extract the cannabinoids or cannabinoid derivatives. After shaking, the plate was centrifuged at 1000 x g for 60 seconds to remove any solids. After centrifugation, 12 pL of supernatant was transferred to a fresh assay plate containing 228 pL of methanol (or more to maintain samples in the assay linear range), sealed with a foil seal, shaken for 60 seconds at 900 rpm, and analyzed by LC-MS.

Analytical methods

[001679] Samples were analyzed by LC-MS mass spectrometer (Agilent 6470) using a Phenomenex Kinetex Phenyl-Hexyl 2.1 x 30 mm, 2.6 pm analytical column with the following gradient (Mobile Phase A: LC-MS grade water with 0.1% formic acid, Mobile Phase B: LC-MS grade acetonitrile with 0.1% formic acid):

[001680] The mass spectrometer was operated in negative ion multiple reaction monitoring mode. Each cannabinoid or cannabinoid derivative was identified by retention time, determined from an authentic standard, and multiple reaction monitoring (MRM) transition:

Recovery and purifications

[001681] Whole-cell broth from cultures comprising modified host cells of the disclosure are extracted with a suitable organic solvent to afford cannabinoids or cannabinoid derivatives. Suitable organic solvents include, but are not limited to, hexane, heptane, ethyl acetate, petroleum ether, and di-ethyl ether, chloroform, and ethyl acetate.

The suitable organic solvent, such as hexane, is added to the whole-cell broth from fermentations comprising modified host cells of the disclosure at a 10: 1 ratio (10 parts whole-cell broth - 1 part organic solvent) and stirred for 30 minutes. The organic fraction is separated and extracted twice with an equal volume of acidic water (pH 2.5). The organic layer is then separated and dried in a concentrator (rotary evaporator or thin film evaporator under reduced pressure) to obtain crude cannabinoid or cannabinoid derivative crystals. The crude crystals may then be heated to 105 °C for 15 minutes followed by 145 °C for 55 minutes to decarboxylate the crude cannabinoid or cannabinoid derivative. The crude crystalline product is re-dissolved and recrystallized in a suitable solvent (e.g., n-pentane) and filtered through a 1 pm filter to remove any insoluble material. The solvent is then removed e.g., by rotary evaporation, to produce pure crystalline product.

In vitro enzyme assay and cell-free production of cannabinoids or cannabinoid derivatives

[001682] In some embodiments, modified host cells, e.g., modified yeast cells are cultured in 96-well microtiter plates containing 360 pL of YPD (10 g/L yeast extract, 20 g/L Bacto peptone, 20 g/L dextrose (glucose)) and sealed with a breathable film seal. Cells are then cultured at 30 °C in a high capacity microtiter plate incubator shaking at 1000 rpm and 80% humidity for 3 days until the cultures reach carbon exhaustion. The growth-saturated cultures are then subcultured into 200 mL of YPGAL media to an OD600 of 0.2 and incubated with shaking for 20 hours at 30 °C. Cells are then harvested by centrifugation at 3000 x g for 5 minutes at 4 °C. Harvested cells are then resuspended in 50 mL buffer (50 mM Tris-HCl, 1 mM EDTA, 0.1 M KC1, pH 7.4, 125 units Benzonase) and then lysed (Emulsiflex C3, Avestin, INC., 60 bar, 10 min). Cells debris is removed by centrifugation (10,000 x g, 10 min, 4 °C). Subsequently, the supernatant is then subjected to

ultracentrifugation (150,000 x g, 1 h, 4 °C, Beckman Coulter L-90K, TI-70). The resulting membrane fractions are then resuspended in 3.3 mL buffer (10 mM Tris-HCl, 10 mM MgCh, pH 8.0, 10% glycerol) and solubilized with a tissue grinder. Then, 0.02% (v/v) of the respective membrane preparations are then dissolved in reaction buffer (50 mM Tris-HCl, 10 mM MgCh, pH 8.5) and substrate (500 pM olivetolic acid, 500 mM GPP) to a total volume of 50 pL and incubated for 1 hour at 30 °C. Assays are then extracted by adding two reaction volumes of ethyl acetate followed by vortexing and centrifugation. The organic layer is evaporated for 30 minutes, resuspended in acetonitrile/H20/formic acid (80:20:0.05%) and filtered with Ultrafree® -MC columns (0.22 pm pore size, PVDF membrane material). Cannabinoids or cannabinoid derivatives are then detected via LC-MS and/or recovered and purified.

Yeast cultivation in a bioreactor

[001683] Single yeast colonies comprising modified host cells disclosed herein are grown in 15 mL of Verduyn medium (originally described by Verduyn et al, Yeast 8(7): 501-17) with 50 mM succinate (pH 5.0) and 2% glucose in a 125 mL flask at 30 °C, with shaking at 200 rpm to an OD600 between 4 to 9. Glycerol is then added to the culture to a concentration of 20% and 1 mL vials of the modified host cell suspension are stored at -80 °C. One to two vials of modified host cells are thawed and grown in Verduyn medium with 50 mM succinate (pH 5.0) and 4% sucrose for 24 hours, then sub-cultured to an OD600 reading of 0.1 in the same media. After 24 hours of growth at 30 °C with shaking, 65 mL of culture is used to inoculate a 1.3-liter fermenter (Eppendorf DASGIP Bioreactor) with 585 mL of Verduyn fermentation media containing 20 g/L galactose supplemented with hexanoic acid (2 mM), a carboxylic acid other than hexanoic acid (2 mM), olivetolic acid (1 mM), or an olivetolic acid derivative (1 mM). A poly-alpha-olefm may be added to the fermenter as an extractive agent. The fermenter is maintained at 30 °C and pH 5.0 with addition of MLOH. In an initial batch phase, the fermenter is aerated at 0.5 volume per volume per minute air (VVM) and agitation ramped to maintain 30% dissolved oxygen. After the initial sugar is consumed, the rise in dissolved oxygen triggers feeding of galactose + hexanoic acid (800 g galactose per liter + 9.28 g hexanoic acid per liter) at 10 g galactose per liter per hour in pulses of 10 g galactose per liter doses (alternatively, rather than feeding the modified host cells disclosed herein hexanoic acid, olivetolic acid, an olivetolic acid derivative, or a carboxylic acid other than hexanoic acid is fed to the modified host cells).

[001684] Between pulses, the feed rate is lowered to 5 g galactose per liter per hour. Upon a 10% rise in dissolved oxygen, the feed rate is resumed at 10 g L^-1 hour^-1. As modified host cell density increases, dissolved oxygen is allowed to reach 0%, and the pulse dose is increased to 50 g galacose per liter. Oxygen transfer rate is maintained at rates representative of full-scale conditions of 100 mM per liter per hour by adjusting agitation as volume increased. Feed rate is adjusted dynamically to meet demand using an algorithm that alternates between a high feed rate and low feed rate. During the low feed rate, modified host cells should consume galactose and hexanoic acid, or, alternatively, olivetolic acid, an olivetolic acid derivative, or a carboxylic acid other than hexanoic acid, and any overflow metabolites accumulated during the high feed rate. A rise in dissolved oxygen triggers the high feed rate to resume. The length of time spent in the low feed rate reflects the extent to which modified host cells are over- or under-fed in the prior high feed rate pulse; this information is then monitored and used to tune the high feed rate up or down, keeping the low feed rate within a defined range.

[001685] Over time, the feed rate matches sugar and hexanoic acid, or, alternatively, olivetolic acid, an olivetolic acid derivative, or a carboxylic acid other than hexanoic acid, demand from modified host cells. This algorithm ensures minimal net accumulation of fermentation products other than cannabinoids or cannabinoid derivatives; biomass; and CO2. In some embodiments, the process continues for 5 to 14 days. In certain such embodiments, accumulated broth is removed daily and assayed for biomass and cannabinoid, or cannabinoid derivative concentration. A concentrated solution of NH4H2PO4, trace metals and vitamins are added periodically to maintain steady state concentrations.

Example 1 : Production of Cannabinoids and Cannabinoid Derivatives with Modified Host Cells

[001686] Data from the assay detailed in the yeast culturing conditions described in the General Methods of the Examples for Saccharomyces cerevisiae strains produced are reported in Tables 2-4. 1 mM olivetolic acid (OA), 2 mM hexanoic acid (HA), or 1 mM of a carboxylic acid other than hexanoic acid were added to the production media (Feed). Table 2: Compilation of CBDA titers in engineered strains

indicated in the“Exp” column. The galactose percentage in each culture was 4% except for in Experiment 7, where it was 2%. For each strain at least 4 colonies were tested and standard deviation is provided.“Fold” was calculated by dividing titer by parent titer from the same experiment. NA = not applicable. ND = no comparison available to calculate Fold. OA = olivetolic acid. Flex = hexanoic acid. S4 is

CEN.PK113-1A with genotype MATalpha; URA3; TRP1 ; LEU2; HIS3; MAL2-8C; SUC2. Neutral integration loci are annotated with an“i” followed by a number and and refer to small deletions in intergenic regions chosen for neutral effect.

Lower case gene names denote deletion of the open reading frame. Promoters are designated with“p” preceding the name of the native gene they regulate. Terminators are omitted here, but shown in the Figures. In reference to OAC* or IRE 1 *,“*” denotes a mutation is present in the sequence. denotes that the experiment was run in defined minimal media (40 g/L galactose, 1 g/L glucose, 15 g/L ammonium sulfate, 8 g/L potassium phosphate, 6.15 g/L magnesium sulfate, 5.9 g/L succinate, 60 pg/L D-biotin, 120 pg/L Ca-D-pantothenate, 120 pg/L nicotinic acid, 300 pg/L myo-inositol, 60 pg/L thiamine hydrochloride, 60 pg/L pyridoxal hydrochloride, 2.5 pg/L p-aminobenzoic acid, 150 mg/L EDTA, 57.5 mg/L zinc sulfate heptahydrate, 2.6 mg/L manganese chloride dihydrate, 4.7 mg/L cobalt chloride hexahydrate, 5 mg/L copper sulfate pentahydrate, 4.8 mg/L sodium molybdate dihydrate, 29 mg/L calcium chloride dihydrate, and 28 mg/L iron sulfate heptahydrate).

Table 3: Compilation of optical densities (ODs) in engineered strains

Table 3 Legend: The galactose percentage in each culture was 4%. Three separate experiments were conducted and each group is indicated in the“Exp” column.

“Fold” was calculated by dividing OD by parent OD from the same experiment. NA = not applicable. OA = olivetolic acid. Neutral integration loci are annotated with an “i” followed by a number and and refer to small deletions in intergenic regions chosen for neutral effect. Lower case gene names denote deletion of the open reading frame. Promoters are designated with“p” preceding the name of the native gene they regulate. Terminators are omitted here, but shown in the Figures.

Table 4: Compilation of CBDA derivatives produced in engineered strains

Example 2: Assay of Expression of Cannabinoid Synthase Polypeptides

[001687] To examine the relationship between synthase codon-optimization and functional expression in naive or secretory pathway modified host strains, strains were constructed containing a CBDAS polypeptide fused to a GFP polypeptide via a cleavable peptide linker, T2A. The CBDAScoX-T2A-GFP cassettes (where X indicates the codon- optimization) express from a single promoter and terminator (pGALl and tTDHl, respectively) but T2A-mediated cleavage results in two separate proteins. In this manner the GFP polypeptide level can be used as a proxy for CBDAS polypeptide level, and the relationships among expression and activity could be investigated. The various CBDAS codon-optimized sequences fused to GFP via T2A were introduced into either a secretory engineering-naive parent (S29) or a secretory engineered parent (S228) and assayed both for GFP polypeptide expression and CBDA titer (Table 5 and FIG. 79).

[001688] Strains containing CBDAScoX-T2A-GFP (in locus i7, from promoter pGALl, and terminator tTDHl; where X refers to codon-optimized sequences 1 (SEQ ID NO:256), 2 (SEQ ID NO:258), 3 (SEQ ID NO:259), 4 (SEQ ID NO:260), 5 (SEQ ID NO:261), 7 (SEQ ID NO:262), or 8 (SEQ ID NO:263) were grown in 96-well plates in non inducing media (360 pL YPD) for two days. Then, these cultures were back-diluted into two plates, one for measuring GFP polypeptide level (360 pL YPG, assayed after 24 hrs of growth) and one for measuring titer (360 pL YPG+olivetolic acid, assayed after 72 hrs of growth). The GFP plate was assayed with a flow analyzer (BD Accuri C6plus) configured for detecting GFP polypeptide. The production plate was assayed with an LC-MS as described under the General Methods of the Examples. Representative strains comprising codon-optimized nucleotide sequences encoding CBDAS polypeptides are shown in Table 5. These codon-optimized nucleotide sequences lack the stop codon, enabling read-through and expression of the GFP polypeptide.

Table 5: Expression level and titer from CBDAS codon-optimizations (codon-opts) fused to GFP via cleavable T2A linker

Table 5 Legend: CBDAS codon-optimized sequences fused to GFP via cleavable T2A linker allow for measurement of CBDAS polypeptide expression level via GFP polypeptide level as a proxy. These cassettes were tested in a non-engineered CBGA producer (S29) or in a secretory-engineered CBGA producer (S228). The data are calculated from four to eight replicate cultures.

[001689] To explore the solubility and subcellular expression of the different synthase codon-optimization variants in naive or secretory-engineered host strains, strains were constructed comprising a CBDAS polypeptide fused to GFP polypeptide via a non-cleavable flexible linker, GS12. The resulting fusion polypeptide can be used to examine how engineering the secretory pathway affects synthase functional expression.

[001690] Strains containing CBDAScoX-GS12-GFP (in locus i7, from promoter pGALl, and terminator tTDHl) were grown in 96-well plates in non-inducing media (360 pL YPD) for two days. Then, these cultures were back-diluted (360 pL YPG) and assayed after 24 hrs of growth. Cultures were washed and resuspended in lx PBS and imaged on a Zeiss LSM 710 confocal system mounted on a Zeiss inverted microscope with a 63 ^c objective and processed using Zeiss Zen software (FIG. 78).

Example 3: Assay of Strains Expressing CBDAS Variant Polypeptides

[001691] Strain S487, described in Table 9, contains all engineering required for production of CBGA from fed olivetolic acid as well as chaperones and secretory pathway engineering features that support expression of CBDAS polypeptides. Variant CBDAS polypeptide constructs were PCR amplified from the selected competition strains and were integrated into S487 using yeast transformation methods described herein. A subset of these constructs were also tested in a strain, S510. Strain S510, described in Table 9, contains all engineering required for production of CBGA from fed olivetolic acid as well as a much more limited set of chaperones and secretory pathway engineering features that support expression of CBDAS polypeptides. Single colonies from the transformations were inoculated directly into the 96 well plate assay. Upon completion, samples were extracted and assayed by LC-MS The data for this assay are shown in Tables 6 and 7.

[001692] Considering strains S699-S791, CBDA titers were improved (outside standard deviation of wild type, S579) in 68 distinct variants covering 53 positions (nearly 10% of all residues). Additionally, some variants displayed a reduction in undesired THCA production. Moreover, growth, as measured by optical density (OD) was improved in most variants. Expression of wild type CBDAS polypeptide makes strains sick, so this is a desirable outcome. A subset of the constructs that improved titers in the S487 background were also tested in S510 background, which contains a more limited set of chaperone and secretory pathway engineering features. Of the 20 mutations tested in this strain, 14 improved titer over the wild type strain SI 100, and six reduced titer. This result indicates that some but not all mutations identified in the S487 background require the support of more extensive chaperone and secretory pathway engineering to function. Table 6: CBDAS Variant Assay (ND = Not Done)

Table 7: CBDAS Variant Assay (n = 4 or greater for all data in Table 7; ND = Not Done)

same experiment control. Four separate experiments were conducted and each group is indicated in the“Exp” column.

** Where multiple strains have the same mutation (e.g., S942, S943), they represent different transformation clones that were compared

Example 4: Assay of Strains Expressing THCAS Variant Polypeptides

[001693] Strain S487, described in Table 9, contains all engineering required for production of CBGA from fed olivetolic acid as well as chaperones and secretory pathway engineering features that support expression of THCAS polypeptides. Variant THCAS polypeptide constructs were PCR amplified from the selected competition strains and were integrated into S487 using yeast transformation methods described herein. Single colonies from the transformations were inoculated directly into the 96 well plate assay Upon completion, samples were extracted and assayed by LC-MS. The results are depicted in Table 8. The control strain here, S1042, contains wild-type THCAS and performs at expected levels. Table 8: THCAS Variants. All strains were assayed at n=3 or greater. NA = not applicable,

ND = not done.

* Absolute values vary by experiment, so values should be compared to their same experiment control. Three separate experiments were conducted and each group is indicated in the“Exp” column.

Table 9: Constructs and strains used in the Examples

In reference to OAC* or IRE1 *, denotes a mutation is present in the sequence.

* If a strain has a parent strain, it is a child strain. All of the constructs present in the parent strain are also all present in the child strain.

** S4 is CEN.PK113-1A with genotype MATalpha; URA3; TRP1; LEU2; HIS3; MAL2-8C; SUC2

*** S228 has no cannabinoid synthase polypeptide but is a parent strain for several strains. **** S369 is a“base strain” with extensive secretory pathway engineering used to test different cannabinoid synthases and codon optimizations. In S369, the nucleotide sequence encoding a GFP polypeptide is added and the synthase polypeptide in parent S270 is deleted, reverting the locus to its wild type state and creating a strain without synthase. Subsequently, the THCAS or other cannabinoid synthase codon optimization variant replaces the nucleotide sequence encoding the GFP polypeptide.

***** S487 and S510 are base strains used to test THCA synthase and CBDA synthase constructs. S487 has extensive chaperone and secretory pathway engineering while S510 has a more minimal set of engineering. In these strains, the nucleotide sequence encoding a pGALl tTDHl empty expression cassette is added as a location for integration of CBDAS variant open reading frames.

Table 10: List of Regulatory and Other Elements

are listed in Table 10. Flanking Homology regions direct recombination at specific genomic loci. Flanking homology upstream sequences are denoted with a“u”, and downstream with a "d"'.“I” indicates an intergenic integration site, e g., ui7, di7 are the regions flanking intergenic region 7. Integrations that delete an open reading frame have flanking homology with the deleted gene indicated, e g., uPEP4, dPEP4 are the regions flanking the PEP4 gene. Synthetic Recombination Sequences (SRS) direct internal recombination of two DNA constructs targeted for integration at the same locus. Linkers are short sequences used in assembly the DNA constructs, they are intervening between the indicated parts. Linkers Gl, G7, G10, RG1, RG10 and LTTDH1 contain the last 36 bp of the upstream DNA part; in cases where these linkers are used assume that the linker reconstitutes sequence omitted from the upstream part to create a seamless junction with the downstream part. Linkers DO and D9 are terminal linkers that direct entry of the DNA constructs into cloning vectors, and are not integrated into the genome. Where no linker is shown between parts, the junction is also seamless.

[001694] Although the present disclosure has been described with reference to the specific embodiments thereof, it should be understood by those skilled in the art that various changes may be made and equivalents may be substituted without departing from the true spirit and scope of the disclosure. In addition, many modifications may be made to adapt a particular situation, material, composition of matter, process, process step or steps, to the objective, spirit and scope of the present disclosure. All such modifications are intended to be within the scope of the claims appended hereto.

Claims

CLAIMS What is claimed is:

1. A modified host cell for producing a cannabinoid or cannabinoid derivative, wherein the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides or a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, or a combination of any of the foregoing, and wherein the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide.

2. The modified host cell of claim 1, wherein the modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides.

3. The modified host cell of claim 2, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin mononucleotide (FMN) synthetase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosidase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosyltransferase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more peptidyl-prolyl isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more fatty acid desaturase polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein transport and trafficking polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more signal peptidase or signal peptidase complex polypeptides, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response (UPR), one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in endoplasmic reticulum-associated degradation (ERAD), one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum, one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cell wall assembly, or one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in vacuolar protein sorting, or a combination of any of the foregoing.

4. The modified host cell of claim 3, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more chaperone or co-chaperone polypeptides.

5. The modified host cell of claim 4, wherein the one or more chaperone or co chaperone polypeptides comprise a KAR2 polypeptide, a JEM1 polypeptide, a LHS1 polypeptide, a SIS1 polypeptide, a SSB1 polypeptide, a CNE1 polypeptide, a CNS1 polypeptide, a PFD2s polypeptide, a PFD1 polypeptide, a SSA1 polypeptide, a YDJ1 polypeptide, a SIL1 polypeptide, a SCJ1 polypeptide, a ROTl polypeptide, a TCP1 polypeptide, a CCT2 polypeptide, a CCT3 polypeptide, a CCT4 polypeptide, a CCT5 polypeptide, a CCT6 polypeptide, a CCT7 polypeptide, a CCT8 polypeptide, a DED1 polypeptide, a CPR5 polypeptide, or a FPR1 polypeptide, or a combination of any of the foregoing.

6. The modified host cell of claim 5, wherein the one or more chaperone or co chaperone polypeptides comprise a JEM1 polypeptide and a KAR2 polypeptide.

7. The modified host cell of claim 5, wherein the one or more chaperone or co chaperone polypeptides comprise a LHS1 polypeptide and a KAR2 polypeptide.

8. The modified host cell of claim 5, wherein the one or more chaperone or co chaperone polypeptides comprise a SIL1 polypeptide and a KAR2 polypeptide.

9. The modified host cell of claim 5, wherein the one or more chaperone or co chaperone polypeptides comprise a CNE1 polypeptide and a KAR2 polypeptide.

10. The modified host cell of claim 5, wherein the one or more chaperone or co chaperone polypeptides comprise a SIS1 polypeptide and a KAR2 polypeptide.

11. The modified host cell of claim 5, wherein the one or more chaperone or co chaperone polypeptides comprise a SSB1 polypeptide and a KAR2 polypeptide.

12. The modified host cell of claim 5, wherein the one or more chaperone or co chaperone polypeptides comprise a ROT1 polypeptide and a KAR2 polypeptide.

13. The modified host cell of claim 5, wherein the one or more chaperone or co chaperone polypeptides comprise a CPR5 polypeptide and a KAR2 polypeptide.

14. The modified host cell of claim 5, wherein the one or more chaperone or co chaperone polypeptides comprise a DED1 polypeptide and a KAR2 polypeptide.

15. The modified host cell of claim 5, wherein the one or more chaperone or co chaperone polypeptides comprise a PFDl polypeptide, a PFD2s polypeptide, and a KAR2 polypeptide.

16. The modified host cell of claim 5, wherein the one or more chaperone or co chaperone polypeptides comprise a KAR2 polypeptide.

17. The modified host cell of any one of claims 5-16, wherein the modified host cell comprises two or more heterologous nucleic acids comprising a nucleotide sequence encoding a KAR2 polypeptide.

18. The modified host cell of any one of claims 5-17, wherein the modified host cell comprises three or more heterologous nucleic acids comprising a nucleotide sequence encoding a KAR2 polypeptide.

19. The modified host cell of claim 5, wherein the one or more chaperone or co chaperone polypeptides comprise a SSB1 polypeptide.

20. The modified host cell of any one of claims 3-19, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in unfolded protein response.

21. The modified host cell of claim 20, wherein the one or more polypeptides involved in unfolded protein response comprise an IRE1 polypeptide, a HACls polypeptide, a DED1 polypeptide, or a PPQ1 polypeptide, or a combination of any of the foregoing.

22. The modified host cell of claim 21, wherein the one or more polypeptides involved in unfolded protein response comprise an IRE1 polypeptide.

23. The modified host cell of claim 21 or 22, wherein the one or more polypeptides involved in unfolded protein response comprise a HACls polypeptide.

24. The modified host cell of any one of claims 21-23, wherein the one or more polypeptides involved in unfolded protein response comprise a PPQ1 polypeptide.

25. The modified host cell of claim 20, wherein the one or more polypeptides involved in unfolded protein response comprise a transcription factor polypeptide or a lumenal sensor polypeptide that binds to an unfolded protein response element.

26. The modified host cell of any one of claims 3-25, wherein the modified host cell comprises a synthetic polypeptide that binds to an unfolded protein response element.

27. The modified host cell of any one of claims 3-26, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in protein translocation into the endoplasmic reticulum.

28. The modified host cell of claim 27, wherein the one or more polypeptides involved in protein translocation into the endoplasmic reticulum comprise a KAR2 polypeptide, a SEC61 polypeptide, a SBH1 polypeptide, a SSS1 polypeptide, a SRP14 polypeptide, a SRP21 polypeptide, a SRP68 polypeptide, a SRP72 polypeptide, a SEC65 polypeptide, a SRP54 polypeptide, a SEC62 polypeptide, a SEC63 polypeptide, a SEC66 polypeptide, a SEC72 polypeptide, a SRP101 polypeptide, or a SRP102 polypeptide, or a combination of any of the foregoing.

29. The modified host cell of claim 28, wherein the one or more polypeptides involved in protein translocation into the endoplasmic reticulum comprise a SEC61 polypeptide, a SBH1 polypeptide, a SSS1 polypeptide, a SRP14 polypeptide, a SRP21 polypeptide, a SRP68 polypeptide, a SRP72 polypeptide, a SEC65 polypeptide, and a SRP54 polypeptide.

30. The modified host cell of claim 28, wherein the one or more polypeptides involved in protein translocation into the endoplasmic reticulum comprise a SRP14 polypeptide, a SRP21 polypeptide, a SRP68 polypeptide, a SRP72 polypeptide, a SEC65 polypeptide, and a SRP54 polypeptide.

31. The modified host cell of any one of claims 27-30, wherein the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding SRP protein complex RNA component SCR1.

32. The modified host cell of any one of claims 3-31, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin adenine dinucleotide (FAD) synthetase polypeptides.

33. The modified host cell of claim 32, wherein the one or more flavin adenine dinucleotide (FAD) synthetase polypeptides comprise a FAD1 polypeptide.

34. The modified host cell of any one of claims 3-33, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more flavin mononucleotide (FMN) synthetase polypeptides.

35. The modified host cell of claim 34, wherein the one or more flavin mononucleotide (FMN) synthetase polypeptides comprise a FMN1 polypeptide.

36. The modified host cell of any one of claims 3-35, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosidase polypeptides.

37. The modified host cell of claim 36, wherein the one or more glycosidase polypeptides comprise a CWH41 polypeptide.

38. The modified host cell of any one of claims 3-37, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more fatty acid desaturase polypeptides.

39. The modified host cell of claim 38, wherein the one or more fatty acid desaturase polypeptides comprise an OLE1 polypeptide.

40. The modified host cell of any one of claims 3-39, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein disulfide isomerase polypeptides.

41. The modified host cell of claim 40, wherein the one or more protein disulfide isomerase polypeptides comprise a PDI1 polypeptide.

42. The modified host cell of any one of claims 3-41, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more thiol oxidase polypeptides.

43. The modified host cell of claim 42, wherein the one or more thiol oxidase polypeptides comprise an EROl polypeptide or an ERV2 polypeptide, or a combination of any of the foregoing.

44. The modified host cell of claim 43, wherein the one or more thiol oxidase polypeptides comprise an EROl polypeptide.

45. The modified host cell of claim 43 or 44, wherein the modified host cell comprises two or more heterologous nucleic acids comprising a nucleotide sequence encoding an EROl polypeptide.

46. The modified host cell of any one of claims 43-45, wherein the one or more thiol oxidase polypeptides comprise an ERV2 polypeptide.

47. The modified host cell of any one of claims 3-46, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more signal peptidase or signal peptidase complex polypeptides.

48. The modified host cell of claim 47, wherein the one or more signal peptidase or signal peptidase complex polypeptides comprise a KEX2 polypeptide, a SPC1 polypeptide, a SPC2 polypeptide, a SPC3 polypeptide, or an SEC11 polypeptide, or a combination of any of the foregoing.

49. The modified host cell of any one of claims 3-48, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more protein transport and trafficking polypeptides.

50. The modified host cell of claim 49, wherein the one or more protein transport and trafficking polypeptides comprise a SEC23 polypeptide, a SEC24 polypeptide, a SEC 13 polypeptide, a BFR2 polypeptide, a SEC31 polypeptide, a SAR1 polypeptide, a SEC12 polypeptide, a SEC4 polypeptide, a SEC 16 polypeptide, an ERV29 polypeptide, a SEC7 polypeptide, a SEC5 polypeptide, a SEC22 polypeptide, a BOS1 polypeptide, a BET1 polypeptide, a BUG 1 polypeptide, a GRH1 polypeptide, a USO 1 polypeptide, a SEC17 polypeptide, a SEC 18 polypeptide, a SSOl polypeptide, a SEC9 polypeptide, a SNC2 polypeptide, or a LDB17 polypeptide, or a combination of any of the foregoing.

51. The modified host cell of claim 50, wherein the one or more protein transport and trafficking polypeptides comprise a BFR2 polypeptide.

52. The modified host cell of claim 50 or 51, wherein the one or more protein transport and trafficking polypeptides comprise a SEC24 polypeptide.

53. The modified host cell of any one of claims 50-52, wherein the one or more protein transport and trafficking polypeptides comprise a LDB17 polypeptide.

54. The modified host cell of any one of claims 3-53, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in endoplasmic reticulum-associated degradation (ERAD).

55. The modified host cell of claim 54, wherein the one or more polypeptides involved in endoplasmic reticulum-associated degradation (ERAD) comprise a NPL4 polypeptide.

56. The modified host cell of any one of claims 3-55, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in cell wall assembly.

57. The modified host cell of claim 56, wherein the one or more polypeptides involved in cell wall assembly comprise a KRE1 polypeptide.

58. The modified host cell of any one of claims 3-57, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more polypeptides involved in vacuolar protein sorting.

59. The modified host cell of claim 58, wherein the one or more polypeptides involved in vacuolar protein sorting comprise a PEPl polypeptide.

60. The modified host cell of any one of claims 3-59, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides comprise one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more glycosyltransferase polypeptides.

61. The modified host cell of any one of claims 1-60, wherein the modified host cell comprises a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides.

62. The modified host cell of claim 61, wherein the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more glycosidase polypeptides, one or more genes encoding one or more glycosyltransferase polypeptides, one or more genes encoding one or more polypeptides involved in lipid droplet assembly, one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism, or one or more genes encoding one or more vacuolar proteinase polypeptides, or a combination of any of the foregoing.

63. The modified host cell of claim 62, wherein the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more glycosidase polypeptides.

64. The modified host cell of claim 63, wherein the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene or a MNS1 gene, or a combination of any of the foregoing.

65. The modified host cell of claim 64, wherein the one or more genes encoding one or more glycosidase polypeptides comprise a ROT2 gene.

66. The modified host cell of any one of claims 62-65, wherein the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more glycosyltransferase polypeptides.

67. The modified host cell of claim 66, wherein the one or more genes encoding one or more glycosyltransferase polypeptides comprise an ALG12 gene.

68. The modified host cell of any one of claims 62-67, wherein the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more polypeptides involved in lipid droplet assembly.

69. The modified host cell of claim 68, wherein the one or more genes encoding one or more polypeptides involved in lipid droplet assembly comprise a FLD1 gene.

70. The modified host cell of any one of claims 62-69, wherein the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism.

71. The modified host cell of claim 70, wherein the one or more genes encoding one or more polypeptides involved in regulation of lipid metabolism comprise an OPI1 gene.

72. The modified host cell of any one of claims 62-71, wherein the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise one or more genes encoding one or more vacuolar proteinase polypeptides.

73. The modified host cell of claim 72, wherein the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene, a PRB1 gene, or a PRC1 gene, or a combination of any of the foregoing.

74. The modified host cell of claim 73, wherein the one or more genes encoding one or more vacuolar proteinase polypeptides comprise a PEP4 gene.

75. The modified host cell of any one of claims 1-74, wherein the modified host cell comprises two or more heterologous nucleic acids comprising a nucleotide sequence encoding the cannabinoid synthase polypeptide.

76. The modified host cell of any one of claims 1-75, wherein the nucleotide sequence encoding the cannabinoid synthase polypeptide is codon-optimized.

77. The modified host cell of any one of claims 1-76, wherein the cannabinoid synthase polypeptide is a tetrahydrocannabinolic acid synthase polypeptide.

78. The modified host cell of any one of claims 1-76, wherein the cannabinoid synthase polypeptide is a cannabichromenic acid synthase polypeptide.

79. The modified host cell of any one of claims 1-76, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide.

80. The modified host cell of claim 79, wherein the cannabidiolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 85% sequence identity to SEQ ID NO:242 or SEQ ID NO:244.

81. The modified host cell of claim 79, wherein the cannabidiolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 90% sequence identity to SEQ ID NO:242 or SEQ ID NO:244.

82. The modified host cell of claim 79, wherein the cannabidiolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 95% sequence identity to SEQ ID NO:242 or SEQ ID NO:244.

83. The modified host cell of claim 79, wherein the cannabidiolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence set forth in SEQ ID NO: 242 or SEQ ID NO: 244.

84. The modified host cell of claim 79, wherein the cannabidiolic acid synthase polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:243 or SEQ ID NO:245.

85. The modified host cell of any one of claims 80-84, wherein the one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide, comprise a nucleotide sequence encoding a signal sequence polypeptide.

86. The modified host cell of any one of claims 1-79, 84, or 85, wherein the cannabinoid synthase polypeptide comprises a signal sequence polypeptide.

87. The modified host cell of claim 86, wherein the signal sequence polypeptide is a secretory signal sequence polypeptide.

88. The modified host cell of claim 87, wherein the secretory signal sequence polypeptide is a native secretory signal sequence polypeptide.

89. The modified host cell of claim 87, wherein the secretory signal sequence polypeptide is a synthetic secretory signal sequence polypeptide.

90. The modified host cell of claim 87, wherein the secretory signal sequence polypeptide is an endoplasmic reticulum retention signal sequence polypeptide.

91. The modified host cell of claim 87, wherein the secretory signal sequence polypeptide is a vacuolar localization signal sequence polypeptide.

92. The modified host cell of claim 91, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide or a PRClt polypeptide.

93. The modified host cell of claim 92, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide.

94. The modified host cell of claim 87, wherein the secretory signal sequence polypeptide is a plasma membrane localization signal sequence polypeptide.

95. The modified host cell of claim 87, wherein the secretory signal sequence polypeptide is a peroxisome targeting signal sequence polypeptide.

96. The modified host cell of claim 79, wherein the cannabidiolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 85% sequence identity to SEQ ID NO: l, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8

97. The modified host cell of claim 79, wherein the cannabidiolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 90% sequence identity to SEQ ID NO: l, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8.

98. The modified host cell of claim 79, wherein the cannabidiolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 95% sequence identity to SEQ ID NO: l, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8.

99. The modified host cell of claim 79, wherein the cannabidiolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence set forth in SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8.

100. The modified host cell of claim 79, wherein the cannabidiolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 85% sequence identity to SEQ ID NO:256, SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO:263.

101. The modified host cell of claim 79, wherein the cannabidiolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 90% sequence identity to SEQ ID NO:256, SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO:263.

102. The modified host cell of claim 79, wherein the cannabidiohc acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 95% sequence identity to SEQ ID NO:256, SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO:263.

103. The modified host cell of claim 79, wherein the cannabidiohc acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence set forth in SEQ ID NO:256, SEQ ID NO:258, SEQ ID N0 259, SEQ ID NO 260, SEQ ID NO:261, SEQ ID NO:262, or SEQ ID NO:263.

104. The modified host cell of claim 79, wherein the cannabidio c acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence set forth in SEQ ID

NO: 389, SEQ ID NO 391, SEQ ID NO:393, SEQ ID NO: 395, SEQ ID NO: 397, SEQ ID NO: 399, SEQ ID NO 401, SEQ ID NO 403, SEQ ID NO 405, SEQ ID NO: 407, SEQ ID NO: 409, SEQ ID NO 411, SEQ ID NO:413, SEQ ID N0 415, SEQ ID NO:417, SEQ ID NO:4l9, SEQ ID NO 421, SEQ ID NO:423, SEQ ID N0 425, SEQ ID NO: 427, SEQ ID NO 429, SEQ ID NO 431, SEQ ID N0 433, SEQ ID NO 435, SEQ ID NO: 437, SEQ ID NO: 439, SEQ ID NO 441, SEQ ID NO:443, SEQ ID N0 445, SEQ ID NO: 447, SEQ ID NO: 449, SEQ ID NO 451, SEQ ID NO:453, SEQ ID N0 455, SEQ ID NO: 457, SEQ ID N0 459, SEQ ID NO 461, SEQ ID N0 463, SEQ ID NO 465, SEQ ID NO: 467, SEQ ID NO: 469, SEQ ID NO 471, SEQ ID NO:473, SEQ ID N0 475, SEQ ID NO: 477, SEQ ID NO: 479, SEQ ID NO 481, SEQ ID NO:483, SEQ ID N0 485, SEQ ID NO:487, SEQ ID NO:489, SEQ ID NO 491, SEQ ID N0 493, SEQ ID N0 495, SEQ ID NO: 497, SEQ ID NO: 499, SEQ ID NO 501, SEQ ID NO:503, SEQ ID NO 505, SEQ ID NO: 507, SEQ ID NO: 509, SEQ ID NO 511, SEQ ID NO:5 l3, SEQ ID N0 515, SEQ ID NO:5l7, SEQ ID NO:5l9, SEQ ID NO 521, SEQ ID N0 523, SEQ ID NO 525, SEQ ID NO: 527, SEQ ID NO: 529, SEQ ID NO 531, SEQ ID NO:533, SEQ ID N0 535, SEQ ID NO: 537, SEQ ID NO: 539, SEQ ID NO 541, SEQ ID NO:543, SEQ ID NO: 545, SEQ ID NO: 547, SEQ ID NO: 549, SEQ ID NO 551, SEQ ID NO:553, SEQ ID N0 555, SEQ ID NO:557, SEQ ID NO:559, SEQ ID NO 561, SEQ ID NO:563, SEQ ID N0 565, SEQ ID NO: 567, SEQ ID NO: 569, SEQ ID NO 571, SEQ ID NO:573, SEQ ID N0 579, SEQ ID NO:58l, SEQ ID NO:583, SEQ ID NO:585, SEQ ID NO:587, SEQ ID N0 589, SEQ ID NO:59l, SEQ ID NO:593, SEQ ID NO:595, SEQ ID NO:597, or SEQ ID NO:599.

105. The modified host cell of claim 77, wherein the tetrahydrocannabinolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence set forth in SEQ ID NO:606, SEQ ID NO:608, SEQ ID NO:6lO, SEQ ID NO:6l2, SEQ ID NO:614, SEQ ID NO:6l6, SEQ ID NO:6l8, SEQ ID NO:620, SEQ ID N0 622, SEQ ID NO:624, SEQ ID NO:626, SEQ ID NO:628, SEQ ID NO:630, SEQ ID N0 632, SEQ ID NO:634, SEQ ID NO:636, SEQ ID NO:638, SEQ ID NO:640, SEQ ID N0 642, SEQ ID NO:644, SEQ ID NO:646, SEQ ID NO:648, SEQ ID NO:650, SEQ ID N0 652, SEQ ID NO:654, SEQ ID NO:656, SEQ ID NO:658, SEQ ID NO 660, SEQ ID N0 662, SEQ ID NO:664, SEQ ID NO:666, SEQ ID NO:668, SEQ ID NO:670, SEQ ID N0 672, SEQ ID NO:674, SEQ ID NO:676, SEQ ID NO:678, SEQ ID NO:680, SEQ ID N0 682, SEQ ID NO:684, SEQ ID NO:686, SEQ ID NO:688, SEQ ID NO 690, SEQ ID N0 692, SEQ ID NO:694, SEQ ID NO:696, SEQ ID NO:698, SEQ ID NO:700, SEQ ID NO:702, SEQ ID NO:704, SEQ ID NO:706, SEQ ID NO:708, SEQ ID NO:7lO, SEQ ID NO:7l2, SEQ ID NO:7l4, SEQ ID NO:716, SEQ ID NO:718, SEQ ID NO 720, SEQ ID N0 722, SEQ ID NO:724, SEQ ID NO:726, SEQ ID NO:728, SEQ ID NO:730, SEQ ID N0 732, SEQ ID NO:734, SEQ ID NO:736, SEQ ID NO:738, SEQ ID NO:740, or SEQ ID NO:742.

106. The modified host cell of claim 77, wherein the tetrahydrocannabinolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 85% sequence identity to SEQ ID N0 238, SEQ ID N0 271, SEQ ID N0 272, SEQ ID N0 273, or SEQ ID NO: 274.

107. The modified host cell of claim 77, wherein the tetrahydrocannabinolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 90% sequence identity to SEQ ID NO:238, SEQ ID NO:27l, SEQ ID NO:272, SEQ ID NO:273, or SEQ ID NO: 274

108. The modified host cell of claim 77, wherein the tetrahydrocannabinolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 95% sequence identity to SEQ ID NO:238, SEQ ID NO:271, SEQ ID NO:272, SEQ ID NO:273, or SEQ ID NO: 274.

109. The modified host cell of claim 77, wherein the tetrahydrocannabinolic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence set forth in SEQ ID NO:238, SEQ ID NO:271, SEQ ID NO:272, SEQ ID N0 273, or SEQ ID NO:274

110. The modified host cell of claim 77, wherein the tetrahydrocannabmolic acid synthase polypeptide is a variant tetrahydrocannabinolic acid synthase polypeptide comprising an amino acid sequence set forth in SEQ ID NO:607, SEQ ID NO:609, SEQ ID NO:6l 1, SEQ ID NO:613, SEQ ID NO:615, SEQ ID NO:617, SEQ ID NO:619, SEQ ID NO:621, SEQ ID

NO: 623, SEQ ID NO: 625, SEQ ID NO: 627, SEQ ID NO: 629, SEQ ID NO: 631, SEQ ID NO:633, SEQ ID NO:635, SEQ ID NO:637, SEQ ID N0 639, SEQ ID NO: 641, SEQ ID NO: 643, SEQ ID NO: 645, SEQ ID NO 647, SEQ ID NO: 649, SEQ ID NO: 651, SEQ ID NO:653, SEQ ID NO:655, SEQ ID NO:657, SEQ ID N0 659, SEQ ID NO: 661, SEQ ID NO:663, SEQ ID NO:665, SEQ ID NO: 667, SEQ ID NO: 669, SEQ ID NO: 671, SEQ ID NO:673, SEQ ID NO:675, SEQ ID NO 677, SEQ ID NO: 679, SEQ ID NO: 681, SEQ ID NO:683, SEQ ID NO:685, SEQ ID NO: 687, SEQ ID NO: 689, SEQ ID NO: 691, SEQ ID NO:693, SEQ ID NO:695, SEQ ID NO: 697, SEQ ID NO: 699, SEQ ID NO: 701, SEQ ID NO:703, SEQ ID NO:705, SEQ ID NO 707, SEQ ID NO: 709, SEQ ID NO:711, SEQ ID NO:713, SEQ ID NO:715, SEQ ID NO:7l7, SEQ ID N0 719, SEQ ID NO: 721, SEQ ID NO: 723, SEQ ID NO: 725, SEQ ID NO: 727, SEQ ID NO 729, SEQ ID NO: 731, SEQ ID N0 733, SEQ ID N0 735, SEQ ID N0 737, SEQ ID N0 739, SEQ ID NO: 741, or SEQ ID NO: 743.

111. The modified host cell of claim 77, wherein the tetrahydrocannabinolic acid synthase polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:239.

112. The modified host cell of claim 78, wherein the cannabichromenic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 85% sequence identity to SEQ ID NO:240.

113. The modified host cell of claim 78, wherein the cannabichromenic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 90% sequence identity to SEQ ID NO:240.

114. The modified host cell of claim 78, wherein the cannabichromenic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence having at least 95% sequence identity to SEQ ID NO:240.

115. The modified host cell of claim 78, wherein the cannabichromenic acid synthase polypeptide is encoded by a codon-optimized nucleotide sequence set forth in SEQ ID NO: 240.

116. The modified host cell of claim 78, wherein the cannabichromenic acid synthase polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:241.

117. The modified host cell of claim 79, wherein the cannabidiolic acid synthase polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 9.

118. The modified host cell of claim 79, wherein the cannabidiolic acid synthase polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:257.

119. The modified host cell of claim 79, wherein the cannabidiolic acid synthase polypeptide is a variant cannabidiolic acid synthase polypeptide comprising an amino acid sequence set forth in SEQ ID NO: 390, SEQ ID NO: 392, SEQ ID NO:394, SEQ ID NO:396,

SEQ ID NO:398, SEQ ID NO:400, SEQ ID NO:402, SEQ ID NO:404, SEQ ID NO:406, SEQ ID NO:408, SEQ ID NO:410, SEQ ID NO:412, SEQ ID NO:4l4, SEQ ID NO:4l6, SEQ ID NO:418, SEQ ID NO:420, SEQ ID NO:422, SEQ ID NO:424, SEQ ID NO:426, SEQ ID NO:428, SEQ ID NO:430, SEQ ID NO:432, SEQ ID NO:434, SEQ ID NO:436, SEQ ID NO:438, SEQ ID NO:440, SEQ ID NO:442, SEQ ID NO:444, SEQ ID NO:446, SEQ ID NO:448, SEQ ID NO:450, SEQ ID NO:452, SEQ ID NO:454, SEQ ID NO:456, SEQ ID NO:458, SEQ ID NO:460, SEQ ID NO:462, SEQ ID NO:464, SEQ ID NO:466, SEQ ID NO:468, SEQ ID NO:470, SEQ ID NO:472, SEQ ID NO:474, SEQ ID NO:476, SEQ ID NO:478, SEQ ID NO:480, SEQ ID NO:482, SEQ ID NO:484, SEQ ID NO:486, SEQ ID NO:488, SEQ ID NO:490, SEQ ID NO:492, SEQ ID NO:494, SEQ ID NO:496, SEQ ID NO:498, SEQ ID NO:500, SEQ ID NO:502, SEQ ID NO:504, SEQ ID NO:506, SEQ ID NO:508, SEQ ID NO:510, SEQ ID NO:512, SEQ ID NO:5l4, SEQ ID NO:5l6, SEQ ID NO:518, SEQ ID NO: 520, SEQ ID NO: 522, SEQ ID NO: 524, SEQ ID NO: 526, SEQ ID NO:528, SEQ ID NO:530, SEQ ID NO:532, SEQ ID NO:534, SEQ ID NO:536, SEQ ID NO:538, SEQ ID NO:540, SEQ ID NO:542, SEQ ID NO:544, SEQ ID NO:546, SEQ ID NO:548, SEQ ID NO:550, SEQ ID NO:552, SEQ ID NO:554, SEQ ID NO:556, SEQ ID NO:558, SEQ ID NO:560, SEQ ID NO:562, SEQ ID NO:564, SEQ ID NO:566, SEQ ID NO:568, SEQ ID NO:570, SEQ ID NO:572, SEQ ID NO:574, SEQ ID NO:580, SEQ ID NO:582, SEQ ID NO:584, SEQ ID NO:586, SEQ ID NO:588, SEQ ID NO:590, SEQ ID NO:592, SEQ ID NO:594, SEQ ID NO:596, SEQ ID NO:598, or SEQ ID N0:600.

120. The modified host cell of any one of claims 1-119, wherein the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate:olivetolic acid geranyltransferase (GOT) polypeptide.

121. The modified host cell of claim 120, wherein the GOT polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:70 or SEQ ID NO:73.

122. The modified host cell of claim 120 or 121, wherein the modified host cell comprises two or more heterologous nucleic acids comprising a nucleotide sequence encoding a GOT polypeptide.

123. The modified host cell of any one of claims 1-119, wherein the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a NphB polypeptide.

124. The modified host cell of claim 123, wherein the NphB polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 576.

125. The modified host cell of any one of claims 1-124, wherein the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a tetraketide synthase (TKS) polypeptide and one or more heterologous nucleic acids comprising a nucleotide sequence encoding an olivetolic acid cyclase (OAC) polypeptide.

126. The modified host cell of claim 125, wherein the TKS polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:75.

127. The modified host cell of claim 125 or 126, wherein the modified host cell comprises three or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide.

128. The modified host cell of any one of claims 125-127, wherein the OAC polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:77 or SEQ ID NO:225

129. The modified host cell of any one of claims 125-128, wherein the modified host cell comprises three or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide.

130. The modified host cell of any one of claims 1-129, wherein the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acyl-activating enzyme (AAE) polypeptide.

131. The modified host cell of claim 130, wherein the AAE polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:79, SEQ ID NO:268, or SEQ ID NO:270.

132. The modified host cell of claim 130 or 131, wherein the modified host cell comprises two or more heterologous nucleic acids comprising a nucleotide sequence encoding an AAE polypeptide.

133. The modified host cell of any one of claims 1-132, wherein the modified host cell comprises one or more of the following: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a HMG-CoA synthase (HMGS) polypeptide; b) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3 -hydroxy-3 -methyl-glutaryl-CoA reductase (tHMGR) polypeptide; c) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate kinase (MK) polypeptide; d) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a phosphomevalonate kinase (PMK) polypeptide; e) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate pyrophosphate decarboxylase (MVD1) polypeptide; or f) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a isopentenyl diphosphate isomerase (IDI1) polypeptide.

134. The modified host cell of claim 133, wherein the IDI1 polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:8l.

135. The modified host cell of claim 133 or 134, wherein the tHMGR polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:83.

136. The modified host cell of any one of claims 133-135, wherein the HMGS polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:85.

137. The modified host cell of any one of claims 133-136, wherein the MK polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:95.

138. The modified host cell of any one of claims 133-137, wherein the PMK polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:93.

139. The modified host cell of any one of claims 133-138, wherein the MVD1 polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:89.

140. The modified host cell of any one of claims 1-139, wherein the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acetoacetyl-CoA thiolase polypeptide.

141. The modified host cell of claim 140, wherein the acetoacetyl-CoA thiolase polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:87.

142. The modified host cell of any one of claims 1-141, wherein the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a pyruvate decarboxylase (PDC) polypeptide.

143. The modified host cell of claim 142, wherein the PDC polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:91.

144. The modified host cell of any one of claims 1-143, wherein the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate synthetase (GPPS) polypeptide.

145. The modified host cell of claim 144, wherein the GPPS polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:97.

146. The modified host cell of claim 1, wherein the modified host cell comprises:

c) two or more heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3-hydroxy-3-methyl-glutaryl-CoA reductase (tHMGR) polypeptide; d) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a mevalonate kinase (MK) polypeptide;

k) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate:olivetolic acid geranyltransferase (GOT) polypeptide; l) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides comprise:

147. The modified host cell of claim 146, wherein the modified host cell comprises: a) two heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide; b) a heterologous nucleic acid comprising a nucleotide sequence encoding a HMG- CoA synthase (HMGS) polypeptide;

c) two heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3 -hydroxy-3 -methyl-glutaryl-CoA reductase (tHMGR) polypeptide;

k) a heterologous nucleic acid comprising a nucleotide sequence encoding a geranyl pyrophosphate: olivetolic acid geranyltransferase (GOT) polypeptide,

l) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides comprise:

3) a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in unfolded protein response, wherein the polypeptide involved in unfolded protein response is an IRE1 polypeptide; and 4) a heterologous nucleic acid comprising a nucleotide sequence encoding a protein disulfide isomerase polypeptide, wherein the protein disulfide isomerase polypeptide is a PDIl polypeptide; and

148. The modified host cell of claim 1, wherein the modified host cell comprises:

j) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate synthetase (GPPS) polypeptide; k) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate:olivetolic acid geranyltransferase (GOT) polypeptide; l) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides comprise:

149. The modified host cell of claim 148, wherein the modified host cell comprises: a) two heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide;

d) a heterologous nucleic acid comprising a nucleotide sequence encoding a mevalonate kinase (MK) polypeptide; e) a heterologous nucleic acid comprising a nucleotide sequence encoding a phosphomevalonate kinase (PMK) polypeptide;

k) a heterologous nucleic acid comprising a nucleotide sequence encoding a geranyl pyrophosphate: olivetolic acid geranyltransferase (GOT) polypeptide;

polypeptides comprise:

1) two heterologous nucleic acids comprising a nucleotide sequence encoding a chaperone or co-chaperone polypeptide, wherein the chaperone or co chaperone polypeptide is a KAR2 polypeptide;

2) a heterologous nucleic acid comprising a nucleotide sequence encoding a polypeptide involved in unfolded protein response, wherein the polypeptide involved in unfolded protein response is an IRE1 polypeptide; and

150. The modified host cell of claim 1, wherein the modified host cell comprises:

c) two or more heterologous nucleic acids comprising a nucleotide sequence encoding a truncated 3-hydroxy-3-methyl-glutaryl-CoA reductase (tHMGR) polypeptide; d) one or more heterologous nucleic acid comprising a nucleotide sequence encoding a mevalonate kinase (MK) polypeptide;

polypeptides comprise:

2) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a protein disulfide isomerase polypeptide, wherein the protein disulfide isomerase polypeptide is a PDI1 polypeptide; and m) deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides, wherein the one or more genes encoding one or more secretory pathway polypeptides deleted or downregulated comprise:

151. The modified host cell of claim 150, wherein the modified host cell comprises: a) two heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide;

polypeptides comprise:

152. The modified host cell of claim 1, wherein the modified host cell comprises:

k) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate:olivetolic acid geranyltransferase (GOT) polypeptide; and l) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides comprise:

153. The modified host cell of claim 152, wherein the modified host cell comprises: a) a heterologous nucleic acid comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide;

k) a heterologous nucleic acid comprising a nucleotide sequence encoding a geranyl pyrophosphate: olivetolic acid geranyltransferase (GOT) polypeptide; and

154. The modified host cell of claim 1, wherein the modified host cell comprises: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide;

k) two or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate olivetolic acid geranyltransferase (GOT) polypeptide; and l) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides comprise:

2) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a chaperone or co-chaperone polypeptide, wherein the chaperone or co-chaperone polypeptide is a KAR2 polypeptide; 3) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a flavin adenine dinucleotide (FAD) synthetase polypeptide, wherein the flavin adenine dinucleotide (FAD) synthetase polypeptide is a FAD1 polypeptide; and

155. The modified host cell of claim 154, wherein the modified host cell comprises: a) a heterologous nucleic acid comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide;

k) two heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate:olivetolic acid geranyltransferase (GOT) polypeptide; and

polypeptides comprise:

156. The modified host cell of claim 1, wherein the modified host cell comprises:

polypeptides comprise:

157. The modified host cell of claim 156, wherein the modified host cell comprises: a) a heterologous nucleic acid comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide; b) a heterologous nucleic acid comprising a nucleotide sequence encoding a HMG- CoA synthase (HMGS) polypeptide;

k) a heterologous nucleic acid comprising a nucleotide sequence encoding a geranyl pyrophosphate: olivetolic acid geranyltransferase (GOT) polypeptide, and

158. The modified host cell of claim 1, wherein the modified host cell comprises:

k) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a geranyl pyrophosphate:olivetolic acid geranyltransferase (GOT) polypeptide; and 1) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides, wherein the one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway

polypeptides comprise:

159. The modified host cell of claim 158, wherein the modified host cell comprises: a) two heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide, wherein the cannabinoid synthase polypeptide is a cannabidiolic acid synthase polypeptide;

j) a heterologous nucleic acid comprising a nucleotide sequence encoding a geranyl pyrophosphate synthetase (GPPS) polypeptide; k) a heterologous nucleic acid comprising a nucleotide sequence encoding a geranyl pyrophosphate: olivetolic acid geranyltransferase (GOT) polypeptide; and

polypeptides comprise:

160. The modified host cell of any one of claims 146-159, wherein the KAR2 polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:23.

161. The modified host cell of any one of claims 146-157, wherein the PDI1 polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:55.

162. The modified host cell of any one of claims 146, 147, or 152-157, wherein the EROl polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:5l .

163. The modified host cell of any one of claims 146-149, wherein the IRE1 polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:67 or SEQ ID NO: 578

164. The modified host cell of any one of claims 156-159, wherein the HACls polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:65.

165. The modified host cell of any one of claims 152-157, wherein the FAD1 polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:37.

166. The modified host cell of claim 156 or 157, wherein the SSB1 polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 13.

167. The modified host cell of any one of claims 146-159, wherein the GOT polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:70 or SEQ ID NO: 73.

168. The modified host cell of any one of claims 146-167, wherein the IDI1 polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:8l .

169. The modified host cell of any one of claims 146-168, wherein the tHMGR polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:83.

170. The modified host cell of any one of claims 146-169, wherein the HMGS polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID N0 85.

171. The modified host cell of any one of claims 146-170, wherein the MK polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:95.

172. The modified host cell of any one of claims 146-171, wherein the PMK polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:93.

173. The modified host cell of any one of claims 146-172, wherein the MVD1 polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:89.

174. The modified host cell of any one of claims 146-173, wherein the acetoacetyl-CoA thiolase polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:87.

175. The modified host cell of any one of claims 146-174, wherein the PDC polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:9l .

176. The modified host cell of any one of claims 146-175, wherein the GPPS polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:97.

177. The modified host cell of any one of claims 146-176, wherein the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding a TKS polypeptide.

178. The modified host cell of claim 177, wherein the TKS polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:75.

179. The modified host cell of any one of claims 146-178, wherein the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an OAC polypeptide.

180. The modified host cell of claim 179, wherein the OAC polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:77 or SEQ ID NO: 225.

181. The modified host cell of any one of claims 146-180, wherein the modified host cell comprises one or more heterologous nucleic acids comprising a nucleotide sequence encoding an acyl-activating enzyme (AAE) polypeptide.

182. The modified host cell of claim 181, wherein the AAE polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO:79, SEQ ID NO:268, or SEQ ID NO:270.

183. The modified host cell of any one of claims 1-182, wherein the modified host cell is a eukaryotic cell.

184. The modified host cell of claim 183, wherein the eukaryotic cell is a yeast cell.

185. The modified host cell of claim 184, wherein the yeast cell is Saccharomyces cerevisiae.

186. The modified host cell of claim 185, wherein the Saccharomyces cerevisiae is a protease-deficient strain of Saccharomyces cerevisiae.

187. The modified host cell of any one of claims 1-186, wherein at least one of the one or more heterologous nucleic acids are integrated into the chromosome of the modified host cell.

188. The modified host cell of any one of claims 1-186, wherein at least one of the one or more heterologous nucleic acids are maintained extrachromosomally.

189. The modified host cell of any one of claims 1-188, wherein at least one of the one or more heterologous nucleic acids are operably-linked to an inducible promoter.

190. The modified host cell of any one of claims 1-188, wherein at least one of the one or more heterologous nucleic acids are operably-linked to a constitutive promoter.

191. The modified host cell of any one of claims 1-190, wherein at least one of the one or more heterologous nucleic acids comprises a codon-optimized nucleotide sequence.

192. The modified host cell of any one of claims 1-191, wherein the modified host cell can produce the cannabinoid or cannabinoid derivative when cultured in a defined medium.

193. The modified host cell of claim 192, wherein the defined medium comprises galactose, glucose, ammonium sulfate, potassium phosphate, magnesium sulfate, succinate, D-biotin, Ca-D-pantothenate, nicotinic acid, myo-inositol, thiamine hydrochloride, pyridoxal hydrochloride, p-aminobenzoic acid, EDTA, zinc sulfate heptahydrate, manganese chloride dihydrate, cobalt chloride hexahydrate, copper sulfate pentahydrate, sodium molybdate dihydrate, calcium chloride dihydrate, and iron sulfate heptahydrate.

194. A method of producing a cannabinoid or a cannabinoid derivative, the method comprising culturing a modified host cell of any one of claims 1-193 in a culture medium.

195. The method of claim 194, wherein the method comprises recovering the produced cannabinoid or cannabinoid derivative.

196. The method of claim 194 or 195, wherein the culture medium comprises a carboxylic acid.

197. The method of claim 196, wherein the carboxylic acid is an unsubstituted or substituted C3-C18 carboxylic acid.

198. The method of claim 197, wherein the unsubstituted or substituted C3-C18 carboxylic acid is an unsubstituted or substituted hexanoic acid.

199. The method of claim 194, wherein the culture medium comprises olivetolic acid or an olivetolic acid derivative.

200. The method of claim 194, wherein the cannabinoid is cannabigerolic acid, cannabigerol, A⁹-tetrahydrocannabinolic acid, A⁹-tetrahydrocannabinol, D⁸- tetrahydrocannabinolic acid, D⁸ -tetrahydrocannabinol, cannabidiolic acid, cannabidiol, cannabichromenic acid, cannabichromene, cannabinolic acid, cannabinol, cannabidivarinic acid, cannabidivarin, tetrahydrocannabivarinic acid, tetrahydrocannabivarin,

201. The method of claim 196, wherein the carboxylic acid is selected from the group consisting of butyric acid, valeric acid, hexanoic acid, octanoic acid, 2-methylhexanoic acid, 3-methylhexanoic acid, 4-methylhexanoic acid, 5-methylhexanoic acid, 2-hexenoic acid, 3- hexenoic acid, 4-hexenoic acid, 5-hexenoic acid, heptanoic acid, 5-chlorovaleric acid, 5- (methylsulfanyl)valeric acid, 4-pentynoic acid, trans-2-pentenoic acid, 5-hexynoic acid, trans-2-hexenoic acid, 6-heptynoic acid, trans-2-octenoic acid, nonanoic acid, trans-2- nonenoic acid, decanoic acid, undecanoic acid, dodecanoic acid, myristic acid, 4- phenylbutyric acid, 5-phenylvaleric acid, 6-phenylhexanoic acid, 7-phenylheptanoic acid, isobutyric acid, fumaric acid, itaconic acid, malic acid, succinic acid, maleic acid, malonic acid, glutaric acid, glucaric acid, oxalic acid, adipic acid, pimelic acid, suberic acid, azelaic acid, sebacic acid, dodecandioic acid, glutaconic acid, ortho-phthalic acid, isophthalic acid, terephthalic acid, citric acid, isocitric acid, aconitic acid, tricarballylic acid, trimesic acid, 5- aminovaleric acid, 5-cyanovaleric acid, 5-hydroxyvaleric acid, and 2,3-dimethylhexanoic acid.

202. The method of claim 196, wherein the carboxylic acid is selected from the group consisting of 2-methylhexanoic acid, 3-methylhexanoic acid, 4-methylhexanoic acid, 5- methylhexanoic acid, 2-hexenoic acid, 3-hexenoic acid, 4-hexenoic acid, heptanoic acid, 5- chlorovaleric acid, 5-(methylsulfanyl)valeric acid, 4-pentynoic acid, trans-2-pentenoic acid, 5-hexynoic acid, trans-2-hexenoic acid, 6-heptynoic acid, trans-2-octenoic acid, nonanoic acid, trans-2-nonenoic acid, decanoic acid, undecanoic acid, dodecanoic acid, myristic acid, 4-phenylbutyric acid, 5 -phenyl valeric acid, 6-phenylhexanoic acid, 7-phenylheptanoic acid, isobutyric acid, fumaric acid, itaconic acid, malic acid, maleic acid, glucaric acid, suberic acid, azelaic acid, sebacic acid, dodecandioic acid, glutaconic acid, ortho-phthalic acid, isophthalic acid, terephthalic acid, citric acid, isocitric acid, aconitic acid, tricarballylic acid, trimesic acid, 5-aminovaleric acid, 5-cyanovaleric acid, 5-hydroxyvaleric acid, and 2,3- dimethylhexanoic acid.

203. The method of claim 196, wherein the carboxylic acid is selected from the group consisting of 4-pentynoic acid, trans-2-pentenoic acid, 5-hexynoic acid, trans-2-hexenoic acid, 6-heptynoic acid, trans-2-octenoic acid, nonanoic acid, trans-2-nonenoic acid, decanoic acid, undecanoic acid, dodecanoic acid, 4-phenylbutyric acid, 5-phenylvaleric acid, 6- phenylhexanoic acid, and 7-phenylheptanoic acid.

204. The method of claim 196, wherein the carboxylic acid is selected from the group consisting of 2-methylhexanoic acid, 4-methylhexanoic acid, 5-methylhexanoic acid, 2- hexenoic acid, 3-hexenoic acid, 4-hexenoic acid, heptanoic acid, 5-chlorovaleric acid, and 5- (methylsulfanyl)valeric acid.

205. The method of claim 196, wherein the carboxylic acid is a carboxylic acid of Formula (II):

wherein R is a Ci-C ixalkyl group substituted with R^a or R is a C2-Ci8alkenyl group substituted with R^a, wherein R^a is -S-Ci-C6alkyl or -S-C2-C6alkenyl.

206. The method of any one of claims 194-205, wherein the culture medium comprises a fermentable sugar.

207. The method of any one of claims 194-205, wherein the culture medium comprises a pretreated cellulosic feedstock.

208. The method of any one of claims 194-205, wherein the culture medium comprises a non-fermentable carbon source.

209. The method of claim 208, wherein the non-fermentable carbon source comprises ethanol.

210. The method of any one of claims 194-209, wherein the culture medium is defined medium.

211. The method of any one of claims 194-209, wherein the defined medium comprises galactose, glucose, ammonium sulfate, potassium phosphate, magnesium sulfate, succinate, D-biotin, Ca-D-pantothenate, nicotinic acid, myo-inositol, thiamine hydrochloride, pyridoxal hydrochloride, p-aminobenzoic acid, EDTA, zinc sulfate heptahydrate, manganese chloride dihydrate, cobalt chloride hexahydrate, copper sulfate pentahydrate, sodium molybdate dihydrate, calcium chloride dihydrate, and iron sulfate heptahydrate.

212. The method of any one of claims 194-211, wherein the cannabinoid or cannabinoid derivative is produced in an amount of more than 100 mg/L culture medium.

213. A method of making a modified host cell of any one of claims 1-193, the method comprising introducing into a host cell: a) one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and b) one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides.

214. A method of making a modified host cell of any one of claims 1-193, the method comprising introducing into a host cell: a) one or more vectors comprising one or more heterologous nucleic acids comprising a nucleotide sequence encoding a cannabinoid synthase polypeptide and b) one or more vectors comprising one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more secretory pathway polypeptides.

215. The method of claim 213 or 214, the method comprising introducing into the host cell a deletion or downregulation of one or more genes encoding one or more secretory pathway polypeptides.

216. A compound of Formula (I):

or a pharmaceutically acceptable salt thereof, wherein,

R¹ is a Ci-Cisalkyl group substituted with R^a or R¹ is a Cb-C ixalkcnyl group substituted with R^a, wherein R^a is -S-Ci-C6alkyl or -S-Cv-Cealkeriyl, or R¹ is selected from the group consisting of:

217. The compound of claim 216, or a pharmaceutically acceptable salt thereof, wherein, R¹ is a Ci-Cisalkyl group substituted with R^a or R¹ is a C2-Cixalkenyl group substituted with R^a, wherein R^a is -S-Ci-C6alkyl or -S-C2-C6alkenyl, or R¹ is selected from the group consisting of:

218. The compound of claim 216, or a pharmaceutically acceptable salt thereof, wherein, R¹ is selected from the group consisting of:

219. The compound of claim 216, or a pharmaceutically acceptable salt thereof, wherein, R¹ is a Ci-Cisalkyl group substituted with R^a or R¹ is a Cb-Cixalkenyl group substituted with R^a, wherein R^a is -S-Ci-C6alkyl or -S-C2-C6alkenyl.

220. The compound of claim 216, or a pharmaceutically acceptable salt thereof, wherein, R¹ is selected from the group consisting of:

221. The compound of claim 216, or a pharmaceutically acceptable salt thereof, wherein the compound of Formula (I) is selected from the group consisting of:

888

889

222. The compound of claim 216, or a pharmaceutically acceptable salt thereof, wherein the compound of Formula (I) is selected from the group consisting of:

223. The compound of claim 216, or a pharmaceutically acceptable salt thereof, wherein the compound of Formula (I) is selected from the group consisting of:

224. A pharmaceutical composition comprising a compound of any one of claims 216- 223, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

225. A method for treating a disorder in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound of any one of claims 216-223, or a pharmaceutically acceptable salt thereof

226. A method for treating a disorder in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of a pharmaceutical composition according to claim 224.

227. The method of claim 225 or 226, wherein the disorder is mediated by cannabinoid receptor type 1 or cannabinoid receptor type 2.

228. The method of claim 225 or 226, wherein the disorder is chronic pain, multiple sclerosis, cancer-associated nausea and vomiting, weight loss, appetite loss, spasticity, or seizures.

229. Use of a compound of any one of claims 216-223, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating a disorder in a subject in need thereof.

230. Use of a pharmaceutical composition according to claim 224 in the manufacture of a medicament for treating a disorder in a subject in need thereof.

231. Use of a compound of any one of claims 216-223, or a pharmaceutically acceptable salt thereof, for treating a disorder in a subject in need thereof.

232. Use of a pharmaceutical composition according to claim 224 for treating a disorder in a subject in need thereof.

233. The use of any one of claims 229-232, wherein the disorder is mediated by cannabinoid receptor type 1 or cannabinoid receptor type 2.

234. The use of any one of claims 229-232, wherein the disorder is chronic pain, multiple sclerosis, cancer-associated nausea and vomiting, weight loss, appetite loss, spasticity, or seizures.

235. A compound of any one of claims 216-223, or a pharmaceutically acceptable salt thereof, for use in a method of treating a disorder in a subject in need thereof.

236. The compound for use of claim 235, wherein the disorder is mediated by cannabinoid receptor type 1 or cannabinoid receptor type 2.

237. The compound for use of claim 235, wherein the disorder is chronic pain, multiple sclerosis, cancer-associated nausea and vomiting, weight loss, appetite loss, spasticity, or seizures.

238. A pharmaceutical composition according to claim 224 for use in a method of treating a disorder in a subject in need thereof.

239. The pharmaceutical composition for use of claim 238, wherein the disorder is mediated by cannabinoid receptor type 1 or cannabinoid receptor type 2.

240. The pharmaceutical composition for use of claim 238, wherein the disorder is chronic pain, multiple sclerosis, cancer-associated nausea and vomiting, weight loss, appetite loss, spasticity, or seizures.

241. A composition comprising a compound of any one of claims 216-223, or a pharmaceutically acceptable salt thereof, and an acceptable carrier.

242. The method of any one of claims 194-211, wherein the cannabinoid or cannabinoid derivative is produced in an amount of more than 50 mg/L culture medium.

243. A nucleic acid comprising a codon-optimized nucleotide sequence having at least 80% sequence identity to SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO: 6, SEQ ID NO: 7, or SEQ ID NO: 8, wherein the codon-optimized nucleotide sequence encodes a cannabidiolic acid synthase polypeptide.

244. The nucleic acid of claim 243, comprising a codon-optimized nucleotide sequence having at least 85% sequence identity to SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8, wherein the codon-optimized nucleotide sequence encodes a cannabidiolic acid synthase polypeptide.

245. The nucleic acid of claim 243, comprising a codon-optimized nucleotide sequence having at least 90% sequence identity to SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8, wherein the codon-optimized nucleotide sequence encodes a cannabidiolic acid synthase polypeptide.

246. The nucleic acid of claim 243, comprising a codon-optimized nucleotide sequence having at least 95% sequence identity to SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8, wherein the codon-optimized nucleotide sequence encodes a cannabidiolic acid synthase polypeptide.

247. A nucleic acid comprising a codon-optimized nucleotide sequence having at least 80% sequence identity to SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO:263, wherein the codon-optimized nucleotide sequence encodes a cannabidiolic acid synthase polypeptide.

248. The nucleic acid of claim 247, comprising a codon-optimized nucleotide sequence having at least 85% sequence identity to SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO:263, wherein the codon- optimized nucleotide sequence encodes a cannabidiolic acid synthase polypeptide.

249. The nucleic acid of claim 247, comprising a codon-optimized nucleotide sequence having at least 90% sequence identity to SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO:263, wherein the codon- optimized nucleotide sequence encodes a cannabidiolic acid synthase polypeptide.

250. The nucleic acid of claim 247, comprising a codon-optimized nucleotide sequence having at least 95% sequence identity to SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:26l, SEQ ID NO:262, or SEQ ID NO:263, wherein the codon- optimized nucleotide sequence encodes a cannabidiolic acid synthase polypeptide.

251. A nucleic acid comprising a codon-optimized nucleotide sequence having at least 80% sequence identity to SEQ ID NO:238, SEQ ID NO:271, SEQ ID NO:272, SEQ ID NO:273, or SEQ ID NO:274, wherein the codon-optimized nucleotide sequence encodes a tetrahydrocannabinolic acid synthase polypeptide.

252. The nucleic acid of claim 251, comprising a codon-optimized nucleotide sequence having at least 85% sequence identity to SEQ ID NO:238, SEQ ID NO:27l, SEQ ID NO:272, SEQ ID NO:273, or SEQ ID NO:274, wherein the codon-optimized nucleotide sequence encodes a tetrahydrocannabinolic acid synthase polypeptide.

253. The nucleic acid of claim 251, comprising a codon-optimized nucleotide sequence having at least 90% sequence identity to SEQ ID NO:238, SEQ ID NO:27l, SEQ ID NO:272, SEQ ID NO:273, or SEQ ID NO:274, wherein the codon-optimized nucleotide sequence encodes a tetrahydrocannabinolic acid synthase polypeptide.

254. The nucleic acid of claim 251, comprising a codon-optimized nucleotide sequence having at least 95% sequence identity to SEQ ID NO:238, SEQ ID NO:271, SEQ ID NO:272, SEQ ID NO:273, or SEQ ID NO:274, wherein the codon-optimized nucleotide sequence encodes a tetrahydrocannabinolic acid synthase polypeptide.

255. A nucleic acid comprising a codon-optimized nucleotide sequence having at least 80% sequence identity to SEQ ID NO:240, wherein the codon-optimized nucleotide sequence encodes a cannabichromenic acid synthase polypeptide.

256. The nucleic acid of claim 255, comprising a codon-optimized nucleotide sequence having at least 85% sequence identity to SEQ ID NO:240, wherein the codon-optimized nucleotide sequence encodes a cannabichromenic acid synthase polypeptide.

257. The nucleic acid of claim 255, comprising a codon-optimized nucleotide sequence having at least 90% sequence identity to SEQ ID NO:240, wherein the codon-optimized nucleotide sequence encodes a cannabichromenic acid synthase polypeptide.

258. The nucleic acid of claim 255, comprising a codon-optimized nucleotide sequence having at least 95% sequence identity to SEQ ID NO:240, wherein the codon-optimized nucleotide sequence encodes a cannabichromenic acid synthase polypeptide.

259. A nucleic acid comprising a codon-optimized nucleotide sequence having at least 80% sequence identity to SEQ ID NO:244, wherein the codon-optimized nucleotide sequence encodes a cannabidiolic acid synthase polypeptide.

260. The nucleic acid of claim 259, comprising a codon-optimized nucleotide sequence having at least 85% sequence identity to SEQ ID NO:244, wherein the codon-optimized nucleotide sequence encodes a cannabidiolic acid synthase polypeptide.

261. The nucleic acid of claim 259, comprising a codon-optimized nucleotide sequence having at least 90% sequence identity to SEQ ID NO:244, wherein the codon-optimized nucleotide sequence encodes a cannabidiolic acid synthase polypeptide.

262. The nucleic acid of claim 259, comprising a codon-optimized nucleotide sequence having at least 95% sequence identity to SEQ ID NO:244, wherein the codon-optimized nucleotide sequence encodes a cannabidiolic acid synthase polypeptide.

263. The nucleic acid of any one of claims 259-262, wherein the nucleic acid comprises a nucleotide sequence encoding a signal sequence polypeptide.

264. The nucleic acid of claim 263, wherein the signal sequence polypeptide is a secretory signal sequence polypeptide.

265. The nucleic acid of claim 264, wherein the secretory signal sequence polypeptide is a native secretory signal sequence polypeptide.

266. The nucleic acid of claim 264, wherein the secretory signal sequence polypeptide is a synthetic secretory signal sequence polypeptide.

267. The nucleic acid of claim 264, wherein the secretory signal sequence polypeptide is an endoplasmic reticulum retention signal sequence polypeptide.

268. The nucleic acid of claim 264, wherein the secretory signal sequence polypeptide is a vacuolar localization signal sequence polypeptide.

269. The nucleic acid of claim 268, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide or a PRClt polypeptide.

270. The nucleic acid of claim 269, wherein the vacuolar localization signal sequence polypeptide is a PEP4t polypeptide.

271. The nucleic acid of claim 264, wherein the secretory signal sequence polypeptide is a plasma membrane localization signal sequence polypeptide.

272. The nucleic acid of claim 264, wherein the secretory signal sequence polypeptide is a peroxisome targeting signal sequence polypeptide.

273. A method of making a modified host cell for producing a cannabinoid or cannabinoid derivative, the method comprising introducing one or more nucleic acids of any one of claims 243-272 into a host cell.

274. A vector comprising one or more nucleic acids of any one of claims 243-272.

275. A method of making a modified host cell for producing a cannabinoid or a cannabinoid derivative, the method comprising introducing one or more vectors of claim 274 into a host cell.

276. A modified host cell for producing a cannabinoid or cannabinoid derivative comprising one or more nucleic acids of any one of claims 243-272.

277. The modified host cell of claim 276, wherein the modified host cell is a eukaryotic cell.

278. The modified host cell of claim 277, wherein the eukaryotic cell is a yeast cell.

279. The modified host cell of claim 278, wherein the yeast cell is Saccharomyces cerevisiae.

280. The modified host cell of claim 279, wherein the Saccharomyces cerevisiae is a protease-deficient strain of Saccharomyces cerevisiae.

281. The modified host cell of any one of claims 276-280, wherein at least one of the one or more nucleic acids are integrated into the chromosome of the modified host cell.

282. The modified host cell of any one of claims 276-280, wherein at least one of the one or more nucleic acids are maintained extrachromosomally.

283. The modified host cell of any one of claims 276-282, wherein at least one of the one or more nucleic acids are operably-linked to an inducible promoter.

284. The modified host cell of any one of claims 276-282, wherein at least one of the one or more nucleic acids are operably-linked to a constitutive promoter.