US20220389467A1

US20220389467A1 - Biosynthetic production of psilocybin and related intermediates in recombinant organisms

Info

Publication number: US20220389467A1
Application number: US17/878,858
Authority: US
Inventors: Jacob Michael Vogan; Laura Flatauer Peiffer; James Lee Wade; Tyrone Jacob Yacoub; Kirsten Tang; Rachel Nadine Burnett
Original assignee: CB Therapeutics Inc USA
Current assignee: CB Therapeutics Inc USA
Priority date: 2019-11-15
Filing date: 2022-08-01
Publication date: 2022-12-08
Also published as: WO2021097452A2; US11441164B2; CA3155976A1; US20210147888A1; WO2021097452A3

Abstract

The systems and methods herein include engineering a host to produce psilocybin using engineered enzymes, genetic changes, and exogenous psilocybin precursor addition (e.g., addition of L-tryptophan to a growing culture of a psilocybin producing recombinant host strain). The process occurs in genetically engineered host cell(s) that can produce psilocybin.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority from U.S. Provisional Application Ser. No. 62/936,387 filed on Nov. 15, 2019, which is incorporated herein by reference in its entirety.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

Not Applicable.

INCORPORATION-BY-REFERENCE OF MATERIAL SUBMITTED ON A COMPACT DISC

The Sequence Listing, which is a part of the present disclosure, includes a computer readable form and a written sequence listing comprising nucleotide and/or amino acid sequences of the present invention. The sequence listing information recorded in computer readable form is identical to the written sequence listing. The ASCII text file, entitled “psilocybinseq.text”, was created on Nov. 15, 2019 using PatentIn version 3.5 and is incorporated herein by reference in its entirety. The subject matter of the Sequence Listing is incorporated herein by reference in its entirety.

FIELD

The present invention generally relates to the production of psilocybin and its intermediates (e.g., tryptamine, 4-hydroxytryptamine, norbaeocystin, baeocystin, and psilocin) in a modified heterologous microorganism.

INTRODUCTION

Mental health problems, which may also be referred to as mental illness or psychiatric disorder, are behavioral or mental patterns which impair the functioning of individuals across the world. Psilocybin has been increasingly evaluated for treating mental health problems. Such mental health disorders include: personality disorders, anxiety disorders, major depressions, and various addictions. In contrast to anxiolytic medicines, usage of psilocybin does not lead to physical dependence.

SUMMARY

The present teachings include a recombinant host organism. The recombinant host organism can include: a plurality of cells transfected by a set of genes for synthesizing psilocybin in the recombinant host organism via at least a first pathway and a second pathway. The recombinant host organism can be a fungal species comprising: Schizosaccharomyces cerevisiae, Schizosaccharomyces japonicus, Schizosaccharomyces pombe, Schizosaccharomyces cryophilus, Saccharomyces cerevisiae, Kluyveromyces lactis, Kluyveromyces dobzhanskii, and Yarrowia lipolytica. The set of genes can include any combination of a gene selected from a group consisting of PsiD, PsiH, PsiK, and PsiM.
In accordance with a further aspect, PsiD can comprise codon optimized nucleic acid sequences SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3 that encode for isolated amino acid sequences SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 16, respectively; PsiH can comprise codon optimized nucleic acid sequences SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6 that encode for isolated amino acid sequences SEQ ID NO: 17 SEQ ID NO: 18, and SEQ ID NO: 19, respectively; PsiK can comprise codon optimized nucleic acid sequences SEQ ID NO: 7 and SEQ ID NO: 8 that encode for isolated amino acid sequences SEQ ID NO: 20 and SEQ ID NO: 21, respectively; and PsiM can comprises codon optimized nucleic acid sequences SEQ ID NO: 9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO: 12, and SEQ ID NO: 13 that encode for isolated amino acid sequences SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24; SEQ ID NO: 25, and SEQ ID NO: 26, respectively.
In accordance with a further aspect, the set of genes can express amino acid sequences that increase titers of psilocybin in the plurality of cells.
In accordance with a further aspect, the set of genes can synthesize intermediates of psilocybin, wherein the intermediates comprise: tryptamine, 4-hydroxytryptamine, norbaeocystin, baeocystin, and psilocin.
In accordance with a further aspect, a protein can be heterologous to the plurality of cells and an exogenous substrate, wherein the protein is encoded by codon optimized SEQ ID NO: 36.
In accordance with a further aspect, the carbon source can include at least one of: glucose, galactose, sucrose, fructose, corn syrup, corn steep liquor, ethanol, and molasses.
In accordance with another aspect, the first pathway can be a shikimate-chorismate pathway and the second pathway can be a L-tryptophan pathway
In accordance with another aspect, the first pathway can be modified by codon optimized SEQ ID NO: 27, SEQ ID NO. 28, SEQ ID NO: 29, SEQ ID NO: 30, and SEQ ID NO: 31 and the second pathway is modified by codon optimized SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, and SEQ ID NO: 35.
The present teaching include a plurality of sequences containing nucleotides or amino acids for producing psilocybin in a recombinant host organism, wherein the plurality of sequences comprise SEQ ID NO: 1-SEQ ID NO: 36.
In accordance with a further aspect, an isolated amino acid sequence comprises SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 16, wherein SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 16 can be at least 50% similar to each other, and wherein SEQ ID NO: 14 is encoded by codon optimized SEQ ID NO: 1, SEQ ID NO: 15 is encoded by codon optimized SEQ ID NO: 2, and SEQ ID NO: 16 is encoded by codon optimized SEQ ID NO: 3.
In accordance with a further aspect, an isolated amino acid sequence comprises at least one of: SEQ ID NO: 17, SEQ ID NO: 18, and SEQ ID NO: 19, wherein SEQ ID NO: 17, SEQ ID NO: 18, and SEQ ID NO: 19 are at least 40% similar to each other, and wherein SEQ ID NO: 17 is encoded by codon optimized SEQ ID NO: 4, SEQ ID NO: 18 is encoded by codon optimized SEQ ID NO: 5, and SEQ ID NO: 19 is encoded by codon optimized SEQ ID NO: 6.
In accordance with a further aspect, an isolated amino acid sequence comprises at least one of: SEQ ID NO: 20 and SEQ ID NO: 21, wherein SEQ ID NO: 20 and SEQ ID NO: 21 are at least 85% similar to each other; and wherein SEQ ID NO: 21 is encoded by codon optimized SEQ ID NO: 7 and SEQ ID NO: 22 is encoded by codon optimized SEQ ID NO: 8.
In accordance with a further aspect, an isolated amino acid sequence comprises at least one of: SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, and SEQ ID NO: 26, wherein SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, and SEQ ID NO: 26 are at least 55% similar to each other, and wherein SEQ ID NO: 22 is encoded by codon optimized SEQ ID NO: 9, SEQ ID NO: 23 is encoded by codon optimized SEQ ID NO: 10, SEQ ID NO: 24 is encoded by SEQ ID NO: 11, SEQ ID NO: 25 is encoded by SEQ ID NO: 12, and SEQ ID NO: 26 is encoded by SEQ ID NO: 13.
The present teachings include a method. The method can include: transfecting a plurality of cells in a recombinant host organism a set of genes for synthesizing psilocybin via at least a first pathway and a second pathway; and increasing titers of psilocybin in the plurality of cells via the set of genes; and synthesizing intermediates of psilocybin via the set of genes. The recombinant host organism can be a fungal species comprising: Schizosaccharomyces cerevisiae, Schizosaccharomyces japonicus, Schizosaccharomyces pombe, Schizosaccharomyces cryophilus, Saccharomyces cerevisiae, Kluyveromyces lactis, Kluyveromyces dobzhanskii, and Yarrowia lipolytica. The The set of genes can include a gene from a group consisting of: PsiD, PsiH, PsiK, and PsiM.
In accordance with a further aspect, PsiD can comprise codon optimized nucleic acid sequences SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3 that encode for isolated amino acid sequences SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 16, respectively; wherein PsiH can comprise codon optimized nucleic acid sequences SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6 that encode for isolated amino acid sequences SEQ ID NO: 17, SEQ ID NO: 18, and SEQ ID NO: 19, respectively; wherein PsiK can comprise codon optimized nucleic acid sequences SEQ ID NO: 7 and SEQ ID NO: 8 that encode for isolated amino acid sequences SEQ ID NO: 20 and SEQ ID NO: 21, respectively; and wherein PsiM can comprise codon optimized nucleic acid sequences SEQ ID NO: 9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO: 12, and SEQ ID NO: 13 that encode for isolated amino acid sequences SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24; SEQ ID NO: 25, and SEQ ID NO: 26, respectively.
In accordance with a further aspect, the carbon source can include at least one of: glucose, galactose, sucrose, fructose, corn syrup, corn steep liquor, ethanol, and molasses.
In accordance with a further aspect, the method can also include an exogenous substrate and a transporter protein.
In accordance with a further aspect, the first pathway can be a shikimate-chorismate pathway modified by codon optimized SEQ ID NO: 27, SEQ ID NO. 28, SEQ ID NO: 29, SEQ ID NO: 30, and SEQ ID NO: 31 and the second pathway can be a L-tryptophan pathway modified by codon optimized SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, and SEQ ID NO: 35.
In accordance with a further aspect, the transporter protein can be encoded by codon optimized SEQ ID NO: 36.
In accordance with a further aspect, the intermediates can include: tryptamine, 4-hydroxytryptamine, norbaeocystin, baeocystin, and psilocin.
These and other features, aspects, and advantages of the present teachings will become better understood with reference to the following description, examples and appended claims.

DRAWINGS

Those of skill in the art will understand that the drawings, described below, are for illustrative purposes only. The drawings are not intended to limit the scope of the present teachings in any way.

FIG. 1 depicts a table of amino acids and codon triplets.

FIG. 2 depicts a table of genes and enzymes inserted into a recombinant host organism.

FIG. 3 depicts the biosynthesis of psilocybin.

FIG. 4-7 depicts sequence alignments.

FIG. 8 depicts endogenous pathways in a host organism.

FIG. 9 depicts a scheme to increase metabolic flux through shikimate-chorismate and L-tryptophan pathways.

FIG. 10 depicts a heterologous recombinant host organism.

FIG. 11 depicts HPLC chromatograms and UV/Vis spectra.

DETAILED DESCRIPTION

Abbreviations and Definitions

To facilitate understanding of the invention, a number of terms and abbreviations as used herein are defined below as follows:
Amino acids: As used herein, the term “amino acids” refer to the molecular basis for constructing and assembling proteins, such as enzymes. (See FIG. 1 for a table of amino acids.). Peptide bonds (i.e., polypeptides) are formed between amino acids and assemble three-dimensionally (3-D). The 3-D assembly can influence the properties, function, and conformational dynamics of the protein. Within biological systems, the protein may: (i) catalyze reactions as enzymes; (ii) transport vesicles, molecules, and other entities within cells as transporter entities; (iii) provide structure to cells and organisms as protein filaments; (iv) replicate deoxyribonucleic acid (DNA); and (v) coordinate actions of cells as cell signalers.
Nucleotides: As used herein, the term “nucleotides” refers to the molecular basis for constructing and assembling nucleic acids, such as DNA and ribonucleic acid (RNA). There are two types of nucleotides—purines and pyrimidines. The specific purines are adenine (A) and guanine (G). The specific pyrimidines are cytosine (C), uracil (U), and thymine (T). T is found in DNA, whereas U is found in RNA. The genetic code defines the sequence of nucleotide triplets (i.e., codons) for specifying which amino acids are added during protein synthesis.
Genes: As used herein, the term “genes” refers to regions of DNA. Amino acid sequences in the proteins, as defined by the sequence of a gene, are encoded in the genetic code.
The present invention is directed to biosynthetic production of psilocybin and related intermediates in recombinant organisms. The syntheses of psilocybin and intermediates of psilocybin in a laboratory environment typically involve tedious techniques of organic chemistry. Often reproducibility is elusive and the solvents used during the syntheses of psilocybin and intermediates of psilocybin are environmentally toxic. Decarboxylations, selective methylations, and selective phosphorylations can be difficult to obtain via the techniques of organic chemistry. Further, the yields and purity of the intermediates for obtaining the target molecules can be low using the techniques of organic chemistry, where the starting molecule is L-tryptophan and the target molecule is psilocybin.
The systems and method herein disclose more environmentally benign processes which can have higher throughputs (i.e., more robust processes). The systems and methods herein include: (i) growing modified recombinant host cells and thereby yielding a recombinant host organism; (ii) expressing engineered psilocybin biosynthesis genes and enzymes in the recombinant host organism; (iii) producing or synthesizing psilocybin and/or intermediates of psilocybin in the recombinant host organism; (iv) fermenting the recombinant host organism; and (v) isolating the psilocybin and/or intermediates of psilocybin from the recombinant host organism. Endogenous pathways of the recombinant host can be modified by the systems and methods herein to produce high purity psilocybin and/or intermediates of psilocybin.
Reference is made to the figures to further describe the systems and methods disclosed herein.
Referring to FIG. 2 , a table lists the enzymes involved in the direct biosynthesis of psilocybin and psilocybin intermediates in species of fungus (i.e., mushrooms). Gene source organisms provide a genetic starting source (i.e., raw gene sequences) which is codon optimized and engineered to function in the recombinant host organisms. The recombinant host organisms include but are not limited to: Schizosaccharomyces cerevisiae, Schizosaccharomyces japonicus, Schizosaccharomyces pombe, Schizosaccharomyces cryophilus, Saccharomyces cerevisiae, Kluyveromyces lactis, Kluyveromyces dobzhanskii, and Yarrowia lipolytica.
Further, the genes/enzymes that are inserted or engineered into the recombinant host are PsiD, PsiH, PsiK, and PsiM.
A PsiD enzyme, which is a decarboxylase (e.g., L-tryptophan decarboxylase) derives from a gene source organism herein—Psilocybe cubensis, Psilocybe cyanescens, and Gymnopilus junonius. The decarboxylase can catalyze the decarboxylation of an aliphatic carboxylic acid (i.e., release carbon dioxide) L-tryptophan to tryptamine and 4-hydroxy-L-tryptophan to 4-hydroxytryptamine, as depicted in FIG. 3 .
A PsiH enzyme, which is a monooxygenase (e.g., Tryptamine 4-monooxygenase) derives from a gene source organism herein—Psilocybe cubensis, Psilocybe cyanescens, and Gymnopilus junonius. The monooxygenase can catalyze the oxidative hydroxylation of the phenyl ring of tryptamine to 4-hydroxytryptamine, as depicted in FIG. 3 .
A PsiK enzyme, which is a kinase (e.g., 4-hydroxytryptamine kinase) derives from a gene source organism herein—Psilocybe cubensis and Psilocybe cyanescens. The kinase can catalyze the phosphorylation (i.e., adding O═P(OH)₂) of the phenolic oxygen of 4-hydroxytryptamine to norbaeocystin, as depicted in FIG. 3 . The kinase can also catalyze the phosphorylation of psilocin to psilocybin.
A PsiM enzyme, which is a methyl transferase (e.g., psilocybin synthase) derives from a gene source organism herein—Psilocybe cubensis, Psilocybe cyanescens, Panaeolus cynascens, Gymnopilus junonius, and Gymnopilus dilepis. The methyl transferase can catalyze the alkylation (i.e., adding a methyl (CH₃) group) of the primary amine in norbaeocystin to baecystin, as depicted in FIG. 3 . Another alkylation can take place where the methyl transferase when the secondary amine of baecystin becomes a tertiary amine of psilocybin, as depicted in FIG. 3 .
As depicted in FIG. 3 , the engineered PsiD, PsiH, PsiK, and PsiM enzymes act on substrates in the psilocybin biosynthetic pathway to produce intermediates of psilocybin and psilocybin itself. The initial substrate for psilocybin intermediates and psilocybin can be L-tryptophan and/or 4-hydroxy-L-tryptophan. These initial substrates can be produced endogenously in a recombinant host as described and/or provided exogenously to a fermentation involving a recombinant host, whereby the host uptakes the starting substrates to feed into the psilocybin biosynthetic pathway. The recombinant host herein described that is expressing all, one, or multiple combinations of the engineered PsiD, PsiH, PsiK, PsiM genes can produce tryptamine, 4-hydroxytryptamine, norbaeocystin, baeocystin, psilocybin, and psilocin. Psilocybin may be converted to psilocin due to spontaneous dephosphorylation. Psilocin is in turn an intermediate which can be acted on by the PsiK enzyme to produce psilocybin.
As depicted in FIG. 4 , the amino acid alignments of recombinant PsiD enzymes are presented. Recombinant PsiD enzymes have been reengineered from various fungal species to function in heterologous recombinant host organisms herein. The gene used in the pair wise alignment is the PsiD gene from the fungal species—Psilocybe cubensis, Psilocybe cyanescens, and Gymnopilus junonius. The alignment is performed with EMBOSS Needle Pair wise Sequence Alignment statistic (EBLOSUM62) with Psilocybe cubensis (PsiD gene) as a reference. The identity percentage and similarity percentage of the amino acid sequences are presented.
For the PsiD gene, codon optimized nucleic acid sequences SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3 encode for isolated amino acid sequences SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 16, respectively. SEQ ID NO: 14 is Psilocybe cubensis (PsiD gene); SEQ ID NO: 15 is Psilocybe cyanescens (PsiD gene); and SEQ ID NO: 16 is Gymnopilus junonius (PsiD gene).
As depicted in FIG. 5 , the amino acid alignment of recombinant PsiH enzymes are presented. Recombinant PsiH enzymes have been reengineered from various fungal species to function in heterologous recombinant host organisms herein. The gene used in the pair wise alignment is the PsiH gene from the fungal species—Psilocybe cubensis, Psilocybe cyanescens, and Gymnopilus junonius. The alignment is performed with EMBOSS Needle Pair wise Sequence Alignment statistic (EBLOSUM62) with Psilocybe cubensis (PsiH gene) as a reference. The identity percentage and similarity percentage of the amino acid sequences are presented.
For the PsiH gene, codon optimized nucleic acid sequences SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6 encode for isolated amino acid sequences SEQ ID NO: 17 SEQ ID NO: 18, and SEQ ID NO: 19, respectively. SEQ ID NO: 17 is Psilocybe cubensis (PsiH gene); SEQ ID NO: 18 is Psilocybe cyanescens (PsiH gene); and SEQ ID NO: 19 is Gymnopilus junonius (PsiH gene).
As depicted in FIG. 6 , the amino acid alignment of recombinant PsiK enzymes are presented. Recombinant PsiK enzymes have been reengineered from various fungal species to function in heterologous recombinant host organisms herein. The gene used in the pair wise alignment is the PsiK gene from the fungal species—Psilocybe cubensis and Psilocybe cyanescens. The alignment is performed with EMBOSS Needle Pair wise Sequence Alignment statistic (EBLOSUM62) with Psilocybe cubensis (PsiK gene) as a reference. The identity percentage and similarity percentage of the amino acid sequences are presented.
For the PsiK gene, codon optimized nucleic acid sequences SEQ ID NO: 7 and SEQ ID NO: 8 encode for isolated amino acid sequences SEQ ID NO: 20 and SEQ ID NO: 21, respectively. SEQ ID NO: 20 is Psilocybe cubensis (PsiK gene) and SEQ ID NO: 21 is Psilocybe cyanescens (PsiK gene).
As depicted in FIG. 7 , the amino acid alignment of recombinant PsiM enzymes are presented. Recombinant PsiM enzymes have been reengineered from various fungal species to function in heterologous recombinant host organisms herein. The gene used in the pair wise alignment is the PsiM gene from the fungal species—Psilocybe cubensis, Psilocybe cyanescens, Panaeolus cynascens, Gymnopilus junonius, and Gymnopilus dilepis. The alignment is performed with EMBOSS Needle Pair wise Sequence Alignment statistic (EBLOSUM62) with Psilocybe cubensis (PsiM gene) as a reference. The identity percentage and similarity percentage of the amino acid sequences are presented.
For the PsiM gene, codon optimized nucleic acid sequences SEQ ID NO: 9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO: 12, and SEQ ID NO: 13 encode for isolated amino acid sequences SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24; SEQ ID NO: 25, and SEQ ID NO: 26, respectively. SEQ ID NO: 22 is Psilocybe cubensis (PsiM gene); SEQ ID NO: 23 is Psilocybe cyanescens (PsiM gene); SEQ ID NO: 24 is Panaeolus cynascens (PsiM gene); SEQ ID NO: 25 is Gymnopilus junonius (PsiM gene), and SEQ ID NO: 26 is Gymnopilus dilepis (PsiM gene).
As depicted in FIG. 8 , the endogenous pathways of a recombinant host organism produce precursors for the engineered PsiD, PsiH, PsiK, PsiM genes. Pathways relating to chorismate, L-glutamine, and L-serine, feed into the endogenous pathway for L-tryptophan production, which a recombinant host organism expressing the psilocybin biosynthetic pathway herein described can use to create tryptamine, 4-hydroxytryptamine, norbaeocystin, baeocystin, psilocin, and psilocybin. The enzymes in the endogenous pathways of the recombinant host organism are encircled in FIG. 8 . Glycolysis and gluconeogenesis in combination with ARO3, ARO4, ARO1, and ARO2 enzymes can be subjected to the depicted precursors at the specified point in the pathway to selectively yield chrorismate. The glutamate biosynthesis pathway in combination with a GLN1 enzyme can be subjected to the depicted precursor at the specified point in the pathway to selectively yield L-glutamine. Glycolysis in combination with SER3, SER33, SER1, and SER2 enzymes can be subjected to the depicted precursors at the specified points in the pathway to selectively yield L-serine. Chorismate and L-glutamine in combination with TRP1, TRP2, TRP3, and TRP4 enzymes can be subjected to the depicted precursors at the specified point to selectively yield (1S,2R)-1-C-indol-3-yl)glycerol 3-phosphate. The addition of L-serine to (1S,2R)-1-C-indol-3-yl)glycerol 3-phosphate in the presence of the TRP1 enzyme can yield L-tryptophan.
As depicted in FIG. 9 , a scheme to increase metabolic flux through the shikimate-chorismate and L-tryptophan pathways is disclosed. The increased metabolic flux through the shikimate-chorismate and L-tryptophan pathways increases the production of L-tryptophan, a key precursor compound for the production of psilocybin and intermediates of psilocybin. Specific enzymes in the described native pathways are overexpressed. Enzymes subject to allosteric inhibition are mutated and overexpressed to render the enzymes insensitive to feedback mechanisms. Enzymes that consume pathway intermediates for off-pathway compound production are hereby deleted.
L-tryptophan production is improved herein by overexpressing a series of enzymes that first increase production of the aromatic compound intermediate, chorismate in a series of enzymatic reactions known as the shikimate pathway. As described in FIG. 5 , the shikimate-chorismate pathway initial precursors, PEP and E4P are converted into 3-deoxy-D-arabinoheptulosonate 7-phosphate (DAHP), catalyzed by ARO3 and ARO4 enzymes.
Overexpression of the genes encoding ARO3 enzyme (as encoded by codon optimized SEQ ID NO: 29), and a feedback-resistant mutant ARO4 K229L enzyme (as encoded by codon optimized SEQ ID NO: 30) are described herein and can increase metabolic flux through the pathway. In addition, genes that encode key enzymes, ARO1 enzyme (as encoded by codon optimized SEQ ID NO: 27) and ARO2 (as encoded by codon optimized SEQ ID NO: 28) are overexpressed as part of a series of enzymes that can convert DAHP to chorismate. In addition, the gene that encodes the Escherichia coli shikimate kinase II (AROL enzyme) can be overexpressed to increase pathway flux from DHAP to chorismate via codon optimized SEQ ID NO: 31.
Chorismate as a general precursor compound can be converted specifically to L-tryptophan by overexpressing a series of enzymes in the L-tryptophan pathway. As described in FIG. 9 , flux through the L-tryptophan pathway can be increased by overexpressing the genes that encode specific enzymes, TRP1 enzyme (as encoded by codon optimized by SEQ ID NO: 32), TRP3 enzyme (as encoded by codon optimized by SEQ ID NO: 34), and TRP4 enzyme (as encoded by codon optimized by SEQ ID NO: 35). Furthermore, overexpression of the gene that encodes the feedback-resistant mutant of TRP2 S76L enzyme (as encoded by SEQ ID NO: 33) is described herein.
Chorismate is a precursor that feeds into the metabolic pathways that produce a variety of aromatic alcohols and aromatic amino acids. The mechanism made operable by systems and methods herein reduce pathway flux into pathways that produce off-pathway targets. As described in FIG. 9 , genes that encode native enzymes—PDC5 enzyme and ARO10 enzyme—have been deleted to reduce pathway flux through the pathways that produce aromatic alcohols. The gene that encodes the native enzyme, ARO7 enzyme has been deleted to reduce production of tyrosine and phenylalanine. Genes that encode PDZ1 and PDZ2 enzymes have been deleted to reduce pathway flux through the pABA production pathway.
As depicted in FIG. 10 , a modified heterologous recombinant host organism is: (i) expressing endogenous pathways for L-tryptophan; (ii) expressing a recombinant version of the TAT2 L-tryptophan importer protein; and (iii) selectively expressing recombinant psilocybin biosynthetic pathway genes. Such a recombinant host can produce tryptamine, 4-hydroxytryptamine, norbaeocystin, baeocystin, psilocin, and psilocybin from L-tryptophan. L-tryptophan can be created by the host through endogenous pathways (FIG. 8 ) or engineered pathways (FIG. 9 ). L-tryptophan may also be fed to the recombinant host organism by media supplementation and up taken by the host expressing the recombinant TAT2 importer protein. Accordingly, contact with the L-tryptophan and the recombinant host organism in the media can selectively direct flux towards psilocybin. Other carbon sources can make contact with the recombinant host organism in the media, wherein the other carbon sources include at least one of: glucose, galactose, sucrose, corn steep liquor, ethanol, fructose, and molasses.
Besides the recombinant TAT2 importer protein, which is encoded by a codon optimized L-tryptophan importer (SEQ ID NO: 36), the nucleotide and amino acid sequences provided are in the order of the psilocybin pathway: PsiD, PsiH, PsiK, and PsiM genes which encode for the respective enzymes. In the systems and methods herein, PsiD enzyme selectively and cleanly catalyzes decarboxylation; the PsiH enzyme catalyzes selective hydroxylation at the 4-position of an indole; the PsiK enzyme catalyzes selective phosphorylation at the hydroxylated 4-position of an indole; and the PsiM enzyme catalyzes selective and stepwise methylations of an amine group, respectively.
By expressing the PsiD gene in the recombinant host organism, codon optimized nucleic acid sequences SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3 encode for isolated amino acid sequences SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 16, respectively. Using the techniques of organic chemistry, decarboxylations would require harsh and toxic tin hydrides (e.g., Barton Decarboxylation), as opposed to the selective and clean decarboxylation by the PsiD enzyme in the recombinant host.
By expressing the PsiH gene in the recombinant host organism, codon optimized nucleic acid sequences SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6 encode for isolated amino acid sequences SEQ ID NO: 17, SEQ ID NO: 18, and SEQ ID NO: 19, respectively. Phenyl group functionalization is often done at high temperatures and pressures, while leading to a mixture of products (e.g., hydroxylations at the 5, 6, and 7 positions of the indole). The regioisomers of the hydroxylated products at the 5, 6, and 7 positions of the indole are structurally distinct from each other, but also structurally similar to each other. Separation of such regioisomers can be very challenging and requires cumbersome separation techniques (e.g., slow column chromatography with poor separation (i.e., the regiosiomers have similar R_fvalues to each other) and low accompanying yields). In contrast, the PsiH enzyme catalyzes selective hydroxylation of indole at the 4-position in the recombinant host organism herein at standard room conditions (˜25 degrees Celsius at ˜1 atm of atmospheric pressure). The systems and methods herein can produce and increase the titers of the hydroxylated indole at the 4-position within the recombinant organism. Using the purification techniques, as described in more detail with respect to the Examples, a sample can be obtained, which exclusively contains the hydroxylated indole at the 4-position. This is indicative of a more facile procedure for obtaining the hydroxylated indole at the 4-position, in comparison to the techniques of organic chemistry.
By expressing the PsiK gene in the recombinant host organism, codon optimized nucleic acid sequences SEQ ID NO: 7 and SEQ ID NO: 8 encode for isolated amino acid sequences SEQ ID NO: 20 and SEQ ID NO: 21, respectively. Primary amines and indole nitrogen are nucleophilic groups than can compete with phenolic oxygen for phosphorylation. In contrast, the recombinant host supports the PsiK enzyme catalysis of selective phosphorylation of the phenolic oxygen. The recombinant host and the PsiK enzyme can also catalyze the undoing of de-phosphorylations that yield psilocin. Stated another way, the recombinant host organism and the expressed PsiK gene for encoding the PsiK enzyme can convert psilocin back to the target molecule psilocybin. Stated yet another way, the recombinant host organism and the expressed PsiK gene for encoding the PsiK enzyme can provide a corrective mechanism to obtain the target molecule psilocybin.
By expressing the PsiM gene in the recombinant host organism, codon optimized nucleic acid sequences SEQ ID NO: 9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO: 12, and SEQ ID NO: 13 encode for isolated amino acid sequences SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, and SEQ ID NO: 26, respectively. The primary amine when subjected to methyl iodide may get over alkylated to the quaternary amine. Further, the reaction is not selective as monoalklyated and dialkylated products may also be obtained. To further complicate the alkylation, the nitrogen of the indole is sufficiently nucleophilic to perform alkylations. In contrast, the PsiM enzyme catalyzes selective methylation at the primary amine in the recombinant host organism, which is also stepwise. The first methylation yields norbaeocystin and the second methylation yields psilocybin. The indole nitrogen does not get methylated.
SEQ ID NO: 1-SEQ ID NO: 36 of the systems and methods herein aid in increasing titers of psilocybin in the recombinant host organism in comparison to the titers of psilocybin in natural state of the host organism. As described above, the mutations at specific points of the pathways above direct flux toward yielding psilocybin in the recombinant host organism.

EXAMPLES

Aspects of the present teachings may be further understood in light of the following examples, which should not be construed as limiting the scope of the present teachings in any way.
The following examples are provided to illustrate various aspects of the present invention. They are not intended to limit the invention, which is defined by the accompanying claims.
In the examples below, genetically engineered host cells may be any species of yeast herein, including but not limited to any species of Saccharomyces, Candida, Schizosaccharomyces, Yarrowia, etc., which have been genetically altered to produce precursor molecules, intermediate molecules, and psilocybin molecules. Additionally, genetically engineered host cells may be any species of filamentous fungus, including but not limited to any species of Aspergillus, which have been genetically altered to produce precursor molecules, intermediate molecules, and psilocybin molecules. Some of the species of yeast herein for the recombinant host organism include but are not limited to: Schizosaccharomyces cerevisiae, Schizosaccharomyces japonicus, Schizosaccharomyces pombe, Schizosaccharomyces cryophilus, Saccharomyces cerevisiae, Kluyveromyces lactis, Kluyveromyces dobzhanskii, and Yarrowia lipolytica.
The gene sequences from gene source organisms are codon optimized to improve expression using techniques disclosed in U.S. patent application Ser. No. 15/719,430, filed Sep. 28, 2017, entitled “An Isolated Codon Optimized Nucleic Acid”. The gene source organisms can include, but are not limited to: Psilocybe cubensis, Psilocybe cyanescens, Panaeolus cynascens, Gymnopilus junonius, and Gymnopilus dilepis. DNA sequences are synthesized and cloned using techniques known in the art. Gene expression can be controlled by inducible or constitutive promoter systems using the appropriate expression vectors. Genes are transformed into an organism using standard yeast or fungus transformation methods to generate modified host strains (i.e., the recombinant host organism). The modified strains express genes for: (i) producing L-tryptophan and precursor molecules to L-tryptophan; (ii) increasing an output of L-tryptophan molecules and precursor molecules to L-tryptophan molecules; (iii) increasing the import of exogenous L-tryptophan into the host strain; and (iv) the genes for the psilocybin biosynthetic pathway. In the presence or absence of exogenous L-tryptophan, fermentations are run to determine if the cell will convert the L-tryptophan into psilocybin. The L-tryptophan and psilocybin pathway genes herein can be integrated into the genome of the cell or maintained as an episomal plasmid. Samples are: (i) prepared and extracted using a combination of fermentation, dissolution, and purification steps; and (ii) analyzed by HPLC for the presence of precursor molecules, intermediate molecules, and psilocybin molecules.
Using the systems and methods herein, the genes which can be expressed to encode for a corresponding enzyme or other type of proteins include but are not limited to: PsiM, PsiH, PsiD, PsiK, TRP1, TRP2 S76L, TRP3, TRP4, ARO1, ARO2, ARO3, ARO4 K229L, and AROL. For example, the PsiM gene is expressed or (overexpressed) to encode for the PsiM enzyme; the PsiH gene is overexpressed to encode for the PsiH enzyme; and so forth. These PsiM, PsiH, PsiD, and PsiK genes can derive from: Psilocybe cubensis, Psilocybe cyanescens, Panaeolus cynascens, Gymnopilus junonius, and Gymnopilus dilepis. These TRP1, TRP2 S76L, TRP3, TRP4, AR01, ARO2, ARO3, and ARO4 K229L genes can derive from Saccharomyces cerevisiae. These AROL genes can derive from Escherichia coli. Further, these genes are transformed into Schizosaccharomyces cerevisiae, Schizosaccharomyces japonicus, Schizosaccharomyces pombe, Schizosaccharomyces cryophilus, Saccharomyces cerevisiae, Kluyveromyces lactis, Kluyveromyces dobzhanskii, and Yarrowia lipolytica. The PsiM, PsiH, PsiD, PsiK, TRP1, TRP2 S76L, TRP3, TRP4, ARO1, ARO2, ARO3, ARO4 K229L, and AROL genes which derive from at least one of: Psilocybe cubensis, Psilocybe cyanescens, Panaeolus cynascens, Gymnopilus junonius, Gymnopilus dilepis, Saccharomyces cerevisiae, and Escherichia coli can be expressed at the same time. Gene sequences can be determined using the techniques disclosed in U.S. Nonprovisional patent application Ser. No. 16/558,909 filed on Sep. 3, 2019, entitled “Automated Pipeline”.

Example 1—Construction of Saccharomyces cerevisiae Platform Strains with Elevated Metabolic Flux Towards L-Tryptophan Via Overexpression of the Feedback Resistant Mutant, ARO4 K229L

The optimized ARO4 K229L gene is synthesized using DNA synthesis techniques known in the art. The optimized gene can be cloned into vectors with the proper regulatory elements for gene expression (e.g. promoter, terminator) and the derived plasmid can be confirmed by DNA sequencing. As an alternative to expression from an episomal plasmid, the optimized ARO4 K229L gene is inserted into the recombinant host genome. Integration is achieved by a single cross-over insertion event of the plasmid. Strains with the integrated gene can be screened by rescue of auxotrophy and genome sequencing.

Example 2—Construction of Saccharomyces cerevisiae Platform Strains with Elevated Metabolic Flux Towards L-Tryptophan Via Deletion of PDC5

Deletion of PDC5 is performed by replacement of the PDC5 gene with the URA3 cassette in the recombinant host. The PDC5 URA3 knockout fragment, carrying the marker cassette, URA3, and homologous sequence to the targeted gene, PDC5, can be generated by bipartite PCR amplification. The PCR product is transformed into a recombinant host and transformants can be selected on synthetic URA drop-out media. Further verification of the modification in said strain can be carried out by genome sequencing, and analyzed by the techniques disclosed in U.S. Nonprovisional patent application Ser. No. 16/558,909 filed on Sep. 3, 2019, entitled “Automated Pipeline”.

Example 3—Method of Growth

Modified host cells that yield recombinant host cells, such as the psilocybin-producing strain herein, express engineered psilocybin biosynthesis genes and enzymes. More specifically, the psilocybin-producing strain herein is grown in rich culture media containing yeast extract, peptone and a carbon source of glucose, galactose, sucrose, fructose, corn syrup, corn steep liquor, ethanol, and/or molasses. The recombinant host cells are grown in either shake flasks or fed-batch bioreactors. Fermentation temperatures can range from 25 degrees Celsius to 37 degrees Celsius at a pH range from pH 4 to pH 7.5. Exogenous L-tryptophan can be added to media to supplement the precursor pool for psilocybin production, which can be up taken by strains expressing the TAT2 L-tryptophan importer protein. The strains herein can be harvested during a fermentation period ranging from 12 hours onward from the start of fermentation.

Example 4—Detection of Isolated Product

To identify fermentation derived psilocybin produced by a recombinant host expressing the engineered psilocybin biosynthetic pathway, an Agilent 1100 series liquid chromatography (LC) system equipped with a HILIC column (Obelisc N, SIELC, Wheeling, Ill. USA) is used. A gradient is used of mobile phase A (ultraviolet (UV) grade H₂O+0.1% Formic Acid) and mobile phase B (UV grade acetonitrile+0.1% Formic Acid). Column temperature is set at 40 degree Celsius. Compound absorbance is measured at 220 nanometers (nm) and 270 nm wavelength using a diode array detector (DAD) and spectral analysis from 200 nm to 400 nm wavelengths. A 0.1 milligram (mg)/milliliter (mL) analytical standard is made from psilocybin certified reference material (Cayman Chemical Company, USA). Each sample is prepared by diluting fermentation biomass from a recombinant host expressing the engineered psilocybin biosynthesis pathway 1:1 in 100% ethanol and filtered in 0.2 um nanofilter vials. Samples are compared to the psilocybin analytical standard retention time and UV-visible spectra for identification. As depicted in inset A of FIG. 11 , a fermentation derived product is obtained which has absorption of 300 au at 220 nm with a retention time of 4.55 minutes in a HPLC chromatogram. As depicted in inset B of FIG. 11 , the fermentation derived product obtained matches the retention time of the psilocybin analytical standard in the overlaid HPLC chromatograms. This indicates that the fermentation derived product is psilocybin. As depicted in inset C of FIG. 11 , the UV-visible spectra of the fermentation derived product and the psilocybin analytical standard are identical. This further corroborates that the fermentation derived product is psilocybin.

OTHER EMBODIMENTS

The detailed description set-forth above is provided to aid those skilled in the art in practicing the present invention. However, the invention described and claimed herein is not to be limited in scope by the specific embodiments herein disclosed because these embodiments are intended as illustration of several aspects of the invention. Any equivalent embodiments are intended to be within the scope of this invention. Indeed, various modifications of the invention in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description which does not depart from the spirit or scope of the present inventive discovery. Such modifications are also intended to fall within the scope of the appended claims.

REFERENCES CITED

All publications, patents, patent applications and other references cited in this application are incorporated herein by reference in their entirety for all purposes to the same extent as if each individual publication, patent, patent application or other reference was specifically and individually indicated to be incorporated by reference in its entirety for all purposes. Citation of a reference herein shall not be construed as an admission that such is prior art to the present invention.

	SEQUENCE LISTINGS
	(Psilocybe cubensis (PSID gene))
	SEQ ID NO: 1
	ATGCAAGTCATCCCCGCGTGCAACAGCGCAGCTAT

	AAGGTCACTTTGTCCGACCCCCGAGAGCTTTAGAA

	ATATGGGCTGGCTTTCCGTGAGCGATGCCGTCTAT

	AGCGAATTTATAGGTGAACTTGCGACGAGAGCATC

	TAATAGAAACTACAGCAATGAGTTCGGTTTAATGC

	AACCAATACAAGAATTTAAAGCGTTCATCGAGAGT

	GATCCCGTTGTACACCAAGAGTTTATCGACATGTT

	TGAAGGCATCCAAGATTCTCCGAGGAACTACCAAG

	AACTATGTAACATGTTCAATGATATTTTTAGGAAG

	GCTCCCGTATACGGAGATTTGGGCCCTCCGGTCTA

	CATGATTATGGCGAAGTTGATGAATACAAGGGCGG

	GTTTCAGTGCGTTCACAAGACAACGTCTGAACCTG

	CATTTTAAAAAGCTGTTCGATACCTGGGGTTTATT

	TCTTTCATCCAAAGACAGCAGGAATGTCCTGGTAG

	CTGACCAGTTTGATGATAGGCACTGCGGCTGGCTG

	AACGAGAGGGCATTATCTGCGATGGTGAAACACTA

	TAATGGGCGTGCATTTGATGAAGTATTTCTATGTG

	ACAAAAATGCACCCTATTACGGCTTTAATTCATAC

	GACGATTTCTTCAATAGGAGGTTCCGTAATAGAGA

	CATTGATAGACCCGTTGTCGGCGGCGTGAACAACA

	CGACGCTTATATCAGCAGCCTGTGAGTCTCTGTCT

	TATAACGTCAGCTATGACGTGCAATCCTTAGATAC

	TTTAGTTTTCAAAGGTGAGACGTACTCATTAAAAC

	ATCTTTTGAATAATGATCCATTTACGCCACAATTC

	GAGCACGGTTCCATATTGCAAGGATTCCTAAACGT

	GACAGCATATCATCGTTGGCACGCGCCGGTTAACG

	GAACTATCGTCAAGATAATCAACGTTCCTGGTACT

	TATTTCGCACAAGCGCCGTCTACCATCGGTGATCC

	GATCCCAGATAATGACTATGATCCACCGCCATATC

	TAAAGAGTCTTGTGTACTTCAGTAACATTGCAGCG

	AGACAGATTATGTTCATAGAAGCTGATAACAAGGA

	GATAGGCCTAATTTTCCTGGTTTTTATAGGCATGA

	CAGAAATTTCAACGTGTGAAGCAACGGTATCCGAG

	GGGCAACATGTCAATAGAGGGGACGACCTGGGTAT

	GTTTCATTTCGGGGGCTCTTCTTTTGCCCTTGGCC

	TGCGTAAAGACTGCCGTGCCGAAATTGTTGAGAAG

	TTCACGGAGCCCGGGACAGTTATAAGGATTAACGA

	AGTCGTCGCCGCCTTGAAGGCTTAA

	(Psilocybe cyanescens (PSID gene))
	SEQ ID NO: 2
	ATGCAAGTGCTTCCTGCTTGCCAAAGCTCTGCCCT

	TAAAACCCTGTGTCCGAGCCCCGAGGCTTTTAGAA

	AGCTGGGATGGCTACCTACGTCTGACGAAGTGTAC

	AACGAGTTCATAGATGATCTGACTGGCAGGACTTG

	CAATGAGAAGTATAGCAGCCAAGTAACCCTGTTAA

	AGCCAATCCAAGACTTCAAGACTTTCATAGAGAAT

	GACCCGATAGTATATCAAGAGTTCATTAGCATGTT

	TGAGGGCATAGAACAGAGCCCTACTAACTATCATG

	AGCTATGTAACATGTTCAACGATATTTTTCGTAAG

	GCACCCCTATACGGAGACTTAGGACCACCTGTCTA

	CATGATAATGGCACGTATTATGAATACGCAGGCGG

	GTTTTTCAGCGTTCACCAAAGAATCTCTGAACTTC

	CATTTTAAGAAGCTATTCGACACGTGGGGTCTATT

	CCTAAGCTCTAAAAATTCCAGAAACGTACTTGTCG

	CCGATCAGTTTGACGACAAACATTACGGATGGTTT

	TCTGAGAGAGCAAAGACTGCGATGATGATCAACTA

	TCCAGGACGTACATTCGAGAAGGTCTTCATCTGTG

	ACGAGCATGTGCCTTATCACGGATTTACTTCCTAT

	GACGACTTCTTTAACAGGAGATTTCGTGACAAGGA

	TACAGACCGTCCCGTCGTCGGTGGCGTCACCGACA

	CGACGTTGATAGGCGCGGCATGTGAAAGTTTATCT

	TATAACGTTTCTCACAACGTCCAATCACTGGACAC

	CCTTGTCATAAAAGGCGAGGCGTACTCTTTAAAAC

	ACCTTCTGCATAATGACCCATTTACGCCACAGTTT

	GAACATGGATCTATCATCCAAGGATTCTTGAACGT

	TACAGCCTATCACAGATGGCACTCTCCAGTTAACG

	GCACTATTGTGAAGATTGTAAACGTACCAGGGACA

	TACTTTGCCCAGGCGCCCTATACCATAGGTAGCCC

	AATCCCTGATAATGACCGGGACCCGCCGCCCTACT

	TGAAGAGCCTTGTTTATTTTAGCAACATTGCTGCC

	AGACAGATTATGTTTATTGAGGCTGACAATAAAGA

	TATTGGCCTTATCTTTCTTGTGTTCATTGGCATGA

	CTGAAATTAGCACATGTGAAGCGACGGTATGCGAA

	GGACAGCACGTTAACAGAGGCGATGACCTTGGGAT

	GTTTCATTTTGGGGGATCGAGTTTTGCATTGGGGC

	TTAGAAAAGATAGCAAAGCAAAAATACTAGAAAAA

	TTTGCAAAGCCGGGAACAGTAATAAGGATTAACGA

	GCTGGTGGCATCCGTCAGAAAATAA

	(Gymnopilus junonius (PSID gene))
	SEQ ID NO: 3
	ATGTCATCTCCTCGTATCGTGCTGCACAGGGTTGG

	TGGCTGGCTGCCTAAAGACCAAAACGTGCTAGAAG

	CATGGCTGAGCAAGAAGATTGCTAAAGCAAAAACT

	AGAAATAGGGCTCCAAAAGATTGGGCTCCTGTGAT

	TCAAGACTTCCAGAGACTGATAGAGACCGATGCCG

	AGATCTACATGGGTTTCCATCAGATGTTCGAGCAG

	GTCCCCAAGAAAACTCCGTACGATAAAGACCCCAC

	CAATGAGCAATGGCAAGTAAGAAATTATATGCACA

	TGTTAGATCTGTTCGACCTAATTATAACCGAGGCA

	CCGGATTTCGAACAAAATGATCTTGTTGGATTTCC

	AATAAATGCAATCCTGGATTGGCCCATGGGGACCC

	CCGGTGGGCTTACTGCATTTATTAACCCTAAAGTA

	AATATTATGTTTCATAAAATGTTTGACGTTTGGGC

	AGTATTTCTGTCATCTCCAGCATCATGCTACGTCC

	TAAATACAAGCGATAGCGGTTGGTTCGGTCCCGCT

	GCAACCGCAGCTATACCCAACTTCAAAGAGACCTT

	CATCTGCGACCCAAGTCTGCCATACCTAGGGTACA

	CTAGCTGGGATAATTTCTTCACCAGGCTGTTTAGG

	CCGGGGGTGCGTCCTGTCGAGTTCCCGAACAATGA

	TGCCATTGTTAACAGTGCGTGTGAATCCACGGTTT

	ATAATATAGCTCCAAACATTAAACCACTAGATAAA

	TTTTGGATTAAGGGAGAGCCGTATTCCCTAAATCA

	CATACTTAATAACGACCCGTACGCGAGCCAGTTCG

	TAGGTGGAACCATATCCCAAGCATTCTTATCTGCG

	CTGAACTATCACCGTTGGGCGAGTCCGGTTAACGG

	CAACATTGTCAAGGTCGTCAATGTTCCGGGTACAT

	ACTACGCGGAGTCCCCAGTTACCGGTTTTGGGAAT

	CCAGAAGGGCCAGATCCAGCGGCGCCCAATCTATC

	TCAAGGTTTCATTACTGCTGTGGCTGCGAGAGCCC

	TGATTTTCATAGAGGCCGATAACCCTAACATCGGA

	TTAATGTGTTTTGTGGGGGTTGGCATGGCAGAGGT

	CTCAACATGTGAAGTTACCGTGAGTGTAGGCGATG

	TTGTCAAGAAAGGAGATGAGATTGGAATGTTCCAT

	TTCGGGGGAAGCACTCACTGCTTGATATTTAGGCC

	ACAAACAAAAATTACGTTCAATCCCGACTATCCTG

	TGTCAACCGCCGTACCCTTGAATGCTGCAGTGGCA

	ACCGTCGTATAA

	(Psilocybe cubensis (PSIH gene))
	SEQ ID NO: 4
	ATGATTGCCGTCTTATTCTCTTTTGTCATAGCTGG

	CTGCATCTATTATATAGTATCCCGTCGTGTGCGTC

	GTTCAAGACTTCCGCCCGGACCACCAGGCATCCCT

	ATCCCCTTTATCGGCAATATGTTTGACATGCCCGA

	AGAATCACCCTGGTTGACGTTTCTGCAATGGGGCA

	GAGATTATAATACAGACATTTTGTATGTAGATGCA

	GGCGGAACTGAGATGGTAATATTGAATACCCTTGA

	GACAATCACTGATTTGTTAGAAAAGAGGGGGTCTA

	TATATTCTGGCAGGCTAGAAAGTACCATGGTTAAT

	GAGTTGATGGGGTGGGAGTTTGATCTAGGATTCAT

	CACCTACGGTGATCGTTGGAGAGAGGAGAGAAGGA

	TGTTCGCGAAAGAGTTCAGCGAAAAGGGAATCAAA

	CAATTCAGGCACGCCCAAGTAAAGGCGGCGCATCA

	ACTTGTCCAACAGCTGACAAAAACACCGGATCGTT

	GGGCTCAACACATACGTCATCAGATAGCCGCCATG

	TCTTTAGACATCGGCTATGGCATAGACTTAGCGGA

	GGATGATCCATGGTTAGAAGCAACACACTTAGCTA

	ACGAAGGACTGGCGATAGCTTCCGTCCCAGGAAAA

	TTTTGGGTAGACTCATTTCCGTCTCTGAAATACCT

	ACCAGCCTGGTTTCCTGGAGCTGTCTTCAAACGTA

	AGGCAAAAGTATGGAGGGAGGCAGCAGACCATATG

	GTGGACATGCCATATGAGACTATGAGGAAATTGGC

	GCCACAGGGCTTGACTAGACCATCCTATGCATCTG

	CAAGACTACAGGCCATGGACCTAAACGGTGATTTG

	GAGCACCAAGAGCACGTAATTAAAAACACAGCAGC

	CGAAGTGAACGTCGGAGGGGGAGATACAACCGTCT

	CTGCGATGAGTGCGTTCATACTAGCGATGGTCAAG

	TATCCGGAAGTACAGCGTAAAGTCCAGGCCGAGCT

	AGACGCACTTACTAACAACGGCCAGATTCCCGATT

	ACGACGAGGAAGACGATAGTCTACCTTACTTGACC

	GCATGTATTAAAGAGTTATTTAGATGGAATCAAAT

	TGCGCCCCTAGCGATTCCTCACAAGTTAATGAAAG

	ACGATGTATATAGGGGTTATCTAATACCTAAGAAT

	ACGCTAGTTTTTGCAAACACATGGGCGGTCCTGAA

	CGACCCTGAAGTCTACCCAGACCCTAGCGTATTTA

	GGCCGGAGCGTTATTTAGGACCCGACGGTAAGCCC

	GATAATACTGTCAGGGACCCCAGGAAGGCTGCGTT

	CGGGTATGGGAGGAGGAACTGTCCAGGAATACACT

	TAGCCCAATCAACCGTCTGGATAGCCGGAGCGACC

	TTACTTAGTGCGTTTAATATCGAGAGGCCAGTTGA

	CCAGAATGGGAAACCCATCGATATTCCAGCAGACT

	TCACAACCGGGTTTTTCAGGCATCCTGTTCCTTTT

	CAGTGCCGTTTCGTGCCTAGGACTGAACAGGTCTC

	CCAATCAGTCAGTGGGCCGTAA

	(Psilocybe cyanescens (PSIH gene))
	SEQ ID NO: 5
	ATGGCGCCTTTGACAACCATGATTCCGATCGTTCT

	ATCTCTTCTAATAGCGGGGTGTATATATTATATCA

	ACGCAAGGAGAATTAAAAGGTCCAGGTTGCCACCA

	GGACCGCCGGGTATTCCTATTCCATTCATCGGGAA

	CATGTTCGACATGCCAAGCGAAAGTCCCTGGCTAA

	TCTTCCTACAATGGGGACAAGAGTACCAGACCGAT

	ATAATTTACGTTGACGCGGGAGGAACTGATATGAT

	AATACTTAATTCCCTAGAGGCAATTACAGATCTGT

	TAGAGAAAAGGGGCTCATTGTATAGCGGGAGGTTG

	GAATCCACGATGGTAAACGAGCTAATGGGTTGGGA

	GTTTGATTTCGGTTTCATACCTTACGGTGAAAGAT

	GGAGGGAAGAACGTCGTATGTTCGCCAAAGAGTTT

	TCTGAGAAGAACATAAGGCAGTTTAGACACGCCCA

	AGTAAAGGCTGCCAATCAGCTAGTGCGTCAACTAA

	CCGATAAACCGGACAGGTGGTCACACCACATAAGG

	CATCAAATCGCGTCCATGGCCCTGGACATCGGTTA

	CGGAATCGATCTTGCTGAAGACGATCCGTGGATCG

	CAGCTTCCGAACTGGCGAATGAAGGCTTGGCTGTA

	GCCTCAGTGCCAGGATCTTTTTGGGTAGATACGTT

	CCCGTTTCTTAAATATTTGCCAAGTTGGTTACCTG

	GCGCGGAGTTCAAAAGAAACGCAAAGATGTGGAAG

	GAAGGAGCAGATCATATGGTCAATATGCCTTACGA

	AACGATGAAAAAGCTAAGCGCACAAGGACTGACTA

	GACCATCATATGCAAGTGCGAGGCTACAGGCTATG

	GACCCGAACGGGGATCTTGAACATCAAGAAAGAGT

	GATCAAAAATACGGCCACGCAGGTAAATGTTGGTG

	GTGGGGATACTACAGTCGGGGCAGTAAGTGCGTTT

	ATCCTTGCGATGGTAAAATACCCGGAAGTTCAAAG

	GAAAGTACAAGCCGAGCTGGACGAGTTCACGAGCA

	AGGGGAGGATACCGGATTACGATGAAGATAACGAT

	TCTCTTCCCTATCTATCGGCTTGCTTCAAAGAGCT

	GTTCAGGTGGGGCCAGATTGCGCCTTTGGCGATTG

	CTCATAGGCTGATAAAGGACGATGTCTATAGGGAA

	TATACTATCCCAAAGAATGCTCTGGTCTTTGCGAA

	CAATTGGTATGGGCGTACTGTATTGAATGACCCTT

	CTGAGTATCCCAATCCTTCAGAATTTAGACCTGAA

	AGGTACTTGGGGCCCGATGGTAAGCCAGATGACAC

	CGTCAGGGACCCAAGAAAGGCAGCGTTTGGGTACG

	GACGTAGAGTGTGTCCAGGGATACACCTGGCGCAG

	AGCACGGTCTGGATTGCTGGTGTCGCGTTGGTATC

	TGCCTTCAACATTGAGCTGCCCGTGGACAAAGACG

	GGAAATGTATAGATATTCCGGCGGCCTTCACGACG

	GGATTCTTTAGATAA

	(Gymnopilus junonius (PSIH gene))
	SEQ ID NO: 6
	ATGATGTCCGAGATGAATGGGATGGATAAATTGGC

	GCTATTGACGACGTTATTAGCTGCCGGTTTTCTAT

	ACTTCAAGAATAAGCGTCGTTCCGCGTTGCCGTTC

	CCGCCAGGGCCGAAAAAGCATCCCCTTTTAGGTAA

	CTTGCTGGACCTTCCGAAGAAGCTGGAGTGGGAGA

	CGTACAGAAGATGGGGAAAAGAATACAATTCAGAT

	GTAATACATGTTAGCGCGGGGAGTGTAAACTTAAT

	TATCGTTAATTCCTTTGAAGCTGCGACAGACCTGT

	TTGATAAGAGATCAGCCAATTATTCAAGTAGGCCA

	CAATTCACGATGGTGAGAGAACTGATGGGATGGAA

	TTGGTTGATGTCTGCATTAATATACGGTGACAAGT

	GGAGAGAGCAACGTAGGTTGTTTCAGAAACATTTC

	AGTACAACGAATGCCGAACTTTACCAAAATACACA

	ATTAGAATATGTTCGTAAAGCCCTGCAGCATCTGC

	TAGAAGAGCCTTCAGATTTTATGGGAATAACACGT

	CACATGGCTGGGGGCGTCAGCATGTCCCTGGCATA

	TGGCTTAAACATTCAGAAGAAAAACGACCCTTTTG

	TTGACCTTGCACAAAGGGCAGTGCACAGCATAACA

	GAGGCCTCAGTTCCTGGGACATTTTGGGTAGACGT

	AATGCCTTGGCTAAAGTATATTCCAGAATGGGTGC

	CGGGTGCTGGCTTTCAGAAGAAGGCTAGAGTGTGG

	AGGAAATTACAGCAAGATTTTCGTCAGGTCCCATA

	TCAGGCAGCTCTGAAAGACATGGCTTCAGGGAAAG

	CTAAACCATCATTTGCAAGTGAGTGTTTGGAGACG

	ATAGACGACAATGAGGATGCACAAAGGCAAAGGGA

	GGTGATAAAAGACACAGCTGCCATTGTATTCGCAG

	CCGGTGCGGATACAAGCCTTAGTGGAATCCATACA

	TTATTCGCCGCAATGTTGTGTTACCCAGAGGTCCA

	GAAGAAAGCACAAGAAGAACTGGATCGTGTCTTGG

	GTGGGAGACGTCTACCGGAATTTACCGATGAGCCC

	AACATGCCCTACATCTCTGCGTTAGTGAAGGAAAT

	ATTGAGGTGGAAACCGGCTACTCCGATTGGCGTAC

	CCCACTTAGCCAGCGAGGATGACGTTTACAACGGA

	TATTACATACCAAAACGTGCGGTTGTCATAGGCAA

	CAGCTGGGCTATGCTTCATGATGAGGAAACTTATC

	CGGACCCAAGCACCTTTAACCCTGACAGATTTTTG

	ACCACAAATAAAAGCACTGGAAAATTGGAATTAGA

	TCCCACAGTGAGAGATCCCGCTTTAATGGCCTTCG

	GATTTGGTAGACGTATGTGTCCAGGACGTGATGTA

	GCTCTTTCTGTCATATGGCTGACTATCGCAAGCGT

	TTTAGCAACGTTTAATATTACCAAGGCGATAGACG

	AAAACGGGAAGGAACTGGAACCGGATGTACAGTAC

	TGGAGCGGTCTAATCGTCCACCCGCTGCCATTCAA

	ATGTACGATCAAGCCAAGATCAAAGGCAGCGGAAG

	AACTTGTGAAATCTGGCGCAGACGCCTATTAA

	(Psilocybe cubensis (PSIK gene))
	SEQ ID NO: 7
	ATGGCATTCGACTTGAAAACTGAAGACGGGCTAAT

	AACTTACCTAACGAAACACCTTTCTTTGGATGTGG

	ATACATCAGGTGTGAAAAGGTTAAGCGGTGGCTTC

	GTTAACGTGACCTGGAGAATAAAACTAAACGCACC

	CTATCAGGGTCACACATCAATAATTCTAAAGCACG

	CACAGCCGCATATGTCAACCGACGAAGACTTCAAA

	ATTGGCGTGGAGCGTTCCGTCTATGAGTACCAGGC

	TATCAAACTTATGATGGCCAATAGGGAGGTGCTAG

	GGGGTGTTGACGGGATCGTGTCTGTGCCAGAGGGG

	TTGAACTACGACCTTGAAAATAATGCATTGATCAT

	GCAGGACGTAGGTAAGATGAAGACCCTATTAGACT

	ACGTAACGGCAAAACCCCCGCTTGCGACTGATATA

	GCACGTTTGGTAGGTACAGAGATTGGGGGTTTCGT

	GGCTAGACTGCATAACATAGGGAGGGAGAGGAGAG

	ACGACCCGGAGTTCAAGTTTTTCTCTGGAAATATA

	GTCGGCAGGACAACAAGCGATCAACTATACCAAAC

	AATTATCCCTAACGCAGCTAAGTACGGGGTAGATG

	ACCCTCTACTGCCTACCGTTGTAAAAGATCTGGTC

	GATGATGTCATGCACAGTGAGGAGACTCTTGTAAT

	GGCGGATTTATGGAGCGGCAATATACTTCTACAGT

	TGGAGGAGGGGAATCCTTCAAAGTTACAGAAAATC

	TACATTTTAGATTGGGAATTGTGTAAATACGGCCC

	AGCTTCACTAGACCTTGGGTATTTCTTGGGTGATT

	GCTACCTGATTTCTCGTTTCCAAGATGAGCAGGTC

	GGCACAACTATGAGACAAGCCTACTTACAAAGCTA

	CGCTCGTACCTCTAAACATTCCATAAACTACGCCA

	AGGTCACTGCGGGAATTGCAGCACATATAGTGATG

	TGGACAGACTTTATGCAGTGGGGGAGTGAGGAAGA

	GAGAATTAACTTCGTCAAGAAAGGCGTGGCCGCCT

	TCCATGACGCAAGAGGGAACAATGATAATGGTGAA

	ATCACCTCTACTCTGTTGAAGGAGAGTTCAACTGC

	CTAA

	(Psilocybe cyanescens (PSIK gene))
	SEQ ID NO: 8
	ATGACTTTCGATCTAAAAACGGAGGAGGGCTTATT

	ATCTTATCTTACCAAGCATTTAAGTTTAGACGTAG

	CACCGAATGGTGTCAAAAGATTATCTGGTGGATTC

	GTCAATGTGACTTGGAGGGTAGGGTTAAATGCACC

	GTACCATGGGCACACGTCTATAATCCTTAAACACG

	CTCAACCACATTTAAGCTCCGATATTGACTTCAAA

	ATAGGGGTGGAAAGAAGTGCGTATGAGTACCAGGC

	TTTGAAGATTGTCTCTGCCAACAGCAGCCTACTTG

	GTTCTTCTGATATCCGTGTCTCAGTTCCAGAAGGT

	TTGCACTATGATGTTGTGAATAACGCCCTAATCAT

	GCAGGACGTGGGTACAATGAAGACCTTGCTGGACT

	ATGTTACAGCGAAACCCCCTATATCTGCTGAAATT

	GCCAGCCTAGTAGGTAGTCAGATTGGCGCTTTCAT

	AGCAAGATTACACAATTTGGGCAGAGAAAATAAAG

	ATAAGGACGACTTTAAATTTTTCTCCGGAAATATA

	GTTGGGAGGACGACGGCAGACCAACTGTATCAGAC

	CATAATTCCTAATGCGGCAAAATATGGAATCGATG

	ACCCAATTCTTCCAATAGTTGTCAAAGAACTTGTT

	GAAGAAGTCATGAACTCAGAGGAAACCCTGATTAT

	GGCGGACCTATGGAGCGGTAATATCTTGCTACAGT

	TCGACGAGAACAGTACGGAACTAACCCGTATTTGG

	CTGGTAGACTGGGAGCTATGCAAGTACGGGCCGCC

	GTCACTGGATATGGGTTACTTCTTGGGCGACTGCT

	TTTTGGTAGCTAGATTCCAAGACCAACTTGTAGGC

	ACATCTATGAGACAAGCATACCTTAAAAGCTACGC

	ACGTAACGTAAAAGAGCCGATCAACTATGCTAAGG

	CCACAGCAGGCATCGGCGCTCATTTGGTAATGTGG

	ACTGACTTCATGAAGTGGGGTAACGATGAAGAAAG

	GGAGGAGTTCGTGAAAAAGGGGGTCGAAGCATTCC

	ACGAGGCCAACGAAGACAATAGGAACGGAGAGATA

	ACGAGCATATTGGTGAAAGAGGCATCACGTACGTA

	A

	(Psilocybe cubensis (PSIM gene))
	SEQ ID NO: 9
	ATGCACATCAGAAACCCCTATAGAACCCCCATAGA

	TTACCAGGCGCTGAGTGAGGCCTTTCCACCATTGA

	AGCCCTTTGTATCCGTAAACGCTGATGGTACGAGT

	TCCGTAGATCTAACGATCCCGGAGGCGCAACGTGC

	GTTCACTGCCGCATTGTTACATAGAGATTTCGGGC

	TAACCATGACTATACCGGAAGATAGACTGTGCCCT

	ACTGTCCCTAACAGGTTAAATTATGTACTGTGGAT

	TGAAGATATTTTCAACTACACGAATAAGACCCTGG

	GGCTGAGCGATGACAGACCGATAAAGGGGGTGGAT

	ATTGGCACAGGCGCCAGCGCAATATACCCTATGCT

	TGCTTGCGCCAGGTTTAAGGCATGGTCCATGGTAG

	GGACAGAGGTAGAACGTAAATGTATTGATACGGCT

	AGACTAAATGTCGTCGCCAATAATCTACAGGATAG

	ATTGAGTATATTAGAGACATCCATCGACGGTCCCA

	TTCTTGTTCCAATCTTCGAGGCCACAGAAGAATAT

	GAGTATGAGTTCACCATGTGTAATCCGCCATTCTA

	CGATGGTGCGGCCGACATGCAGACCTCTGACGCGG

	CCAAAGGATTCGGCTTTGGAGTGGGGGCCCCTCAC

	TCTGGAACAGTTATCGAAATGTCCACTGAAGGAGG

	GGAGTCCGCATTCGTAGCCCAGATGGTGAGAGAGA

	GCTTGAAACTGCGTACCAGATGCAGATGGTATACG

	TCTAATCTTGGGAAATTAAAAAGCCTAAAGGAGAT

	TGTGGGTCTTTTAAAAGAGCTGGAGATTTCCAACT

	ACGCCATAAACGAGTACGTCCAAGGGTCTACCAGA

	AGATACGCCGTCGCGTGGTCTTTTACTGACATTCA

	GCTTCCAGAGGAGCTATCTCGTCCCAGTAACCCGG

	AATTGTCCTCCTTGTTTTAA

	(Psilocybe cyanescens (PSIM gene))
	SEQ ID NO: 10
	ATGCATATCAGGAATCCGTACCGTGACGGCGTGGA

	CTACCAGGCATTAGCCGAGGCTTTCCCGGCGCTAA

	AGCCACACGTCACTGTCAATTCAGACAATACAACT

	TCTATAGATTTCGCGGTACCCGAGGCCCAGAGACT

	TTACACCGCAGCATTACTTCATAGGGACTTTGGTT

	TAACCATAACCTTACCCGAGGATAGACTATGTCCT

	ACGGTCCCGAATAGATTGAACTATGTGTTGTGGGT

	GGAAGATATACTGAAGGTTACGTCAGACGCATTGG

	GATTACCGGATAATAGACAAGTGAAAGGTATTGAT

	ATTGGAACAGGAGCAAGCGCAATTTATCCCATGTT

	AGCTTGTTCCAGGTTTAAGACTTGGTCCATGGTAG

	CTACAGAGGTGGATCAAAAATGCATAGATACCGCA

	AGGCTAAACGTAATAGCTAATAACCTTCAGGAGAG

	ATTGGCAATCATAGCCACTTCCGTGGACGGGCCTA

	TTCTTGTTCCTCTGTTGCAGGCTAATTCCGACTTT

	GAATATGACTTCACCATGTGCAATCCGCCCTTTTA

	CGACGGCGCCTCTGATATGCAGACAAGTGATGCCG

	CTAAAGGCTTTGGCTTCGGAGTAAACGCACCTCAC

	ACTGGGACAGTACTTGAAATGGCGACAGAAGGAGG

	GGAAAGTGCGTTCGTTGCCCAAATGGTTCGTGAGT

	CCTTGAACCTGCAGACTAGATGCAGGTGGTTCACA

	TCTAATTTGGGTAAACTAAAATCACTGTACGAGAT

	TGTGGGTCTATTAAGAGAACACCAGATTTCTAACT

	ACGCCATAAATGAGTATGTACAAGGCGCAACTCGT

	AGGTATGCAATTGCGTGGAGTTTCATAGATGTAAG

	ACTGCCCGACCATTTGTCCAGACCATCTAATCCCG

	ATCTATCCAGTTTGTTTTAA

	(Panaeolus cyanescens (PSIM gene))
	SEQ ID NO: 11
	ATGCATAACCGTAACCCGTATAGGGACGTGATTGA

	TTACCAAGCACTTGCGGAAGCCTACCCGCCCCTAA

	AACCCCACGTCACGGTGAACGCGGATAACACGGCA

	TCCATAGATCTTACGATCCCCGAGGTCCAGAGGCA

	ATACACAGCAGCTCTTTTACATCGTGATTTCGGAT

	TAACTATCACACTACCAGAAGATAGGCTGTGCCCG

	ACAGTACCGAACCGTTTAAACTATGTATTGTGGAT

	AGAGGATATATTTCAGTGTACGAATAAGGCTCTGG

	GATTGTCAGATGACAGACCCGTTAAGGGGGTAGAT

	ATAGGGACCGGCGCCTCCGCCATCTATCCAATGCT

	TGCTTGCGCGAGGTTTAAGCAGTGGTCCATGATTG

	CCACAGAAGTGGAGCGTAAGTGCATAGATACAGCG

	AGATTGAATGTCCTGGCGAATAACTTACAGGACCG

	TTTGTCAATTCTTGAGGTTTCAGTAGACGGCCCGA

	TTTTGGTACCCATCTTTGATACCTTCGAGCGTGCG

	ACAAGCGATTACGAATTTGAGTTCACGATGTGTAA

	CCCTCCATTTTACGACGGGGCCGCGGATATGCAAA

	CATCAGATGCAGCTAAGGGTTTCGGTTTTGGAGTT

	AACGCTCCACACTCCGGTACCGTGATAGAGATGGC

	TACTGAAGGAGGTGAGGCTGCTTTTGTGGCGCAAA

	TGGTCCGTGAGAGCATGAAGTTACAGACAAGGTGT

	CGTTGGTTTACAAGCAACTTAGGCAAGCTAAAATC

	ACTGCATGAAATTGTTGCTTTGTTGAGAGAATCCC

	AGATCACAAACTATGCCATAAATGAGTACGTTCAG

	GGGACGACGAGAAGGTACGCTCTTGCTTGGTCCTT

	CACAGACATAAAACTTACTGAGGAACTTTACAGGC

	CCTCCAATCCAGAATTAGGACCTCTTTGCAGCACA

	TTTGTCTAA

	(Gymnopilus dilepis (PSIM gene))
	SEQ ID NO: 12
	ATGCACATTAGAAACCCTTACTTAACACCTCCGGA

	CTACGAGGCCCTTGCGGAGGCCTTCCCCGCACTAA

	AGCCTTATGTTACAGTTAACCCCGATAAGACTACT

	ACAATTGACTTTGCCATACCGGAGGCTCAGAGATT

	ATACACGGCTGCTCTACTTTACAGGGACTTTGGAC

	TGACAATAACATTGCCGCCGGATAGGTTATGCCCA

	ACCGTGCCCAATAGGCTTAATTATGTTTTGTGGAT

	TCAGGACATTCTGCAGATTACCTCCGCTGCCTTGG

	GCTTGCCAGAGGCTAGACAAGTAAAGGGAGTAGAC

	ATAGGTACCGGAGCGGCAGCGATATACCCTATTCT

	TGGTTGCAGCCTTGCAAAGAATTGGTCTATGGTGG

	GGACAGAGGTCGAACAAAAATGTATCGACATAGCG

	CGTCAAAACGTGATTTCAAATGGATTGCAGGATAG

	GATAACCATAACTGCTAATACCATAGACGCTCCCA

	TTCTGCTGCCCTTATTTGAAGGAGACAGTAACTTC

	GAATGGGAGTTCACCATGTGTAACCCGCCATTTTA

	CGACGGCGCTGCGGACATGGAGACAAGCCAGGACG

	CTAAAGGCTTCGGGTTCGGCGTCAACGCCCCGCAT

	ACAGGAACAGTGGTGGAAATGGCCACGGACGGTGG

	TGAGGCTGCATTCGTCAGCCAAATGGTGAGAGAGT

	CCTTGCACCTAAAGACACGTTGTAGATGGTTCACG

	TCCAATCTAGGTAAATTGAAGAGTCTACATGAAAT

	TGTGGGATTGTTGCGTGAACACCAAATTACCAACT

	ACGCGATAAATGAATATGTTCAGGGAACGACACGT

	AGATACGCGATTGCATGGTCATTTACTGACCTACG

	TCTATCAGACCACCTGCCACGTCCTCCGAACCCCG

	ATCTATCAGCCCTATTTTAA

	(Gymnopilus junonius (PSIM gene))
	SEQ ID NO: 13
	ATGCACTCTCGTAACCCTTATAGATCCCCTCCTGA

	TTTCGCGGCATTAAGTGCGGCTTATCCTCCGCTGT

	CACCATACATAACTACCGATCTAAGCAGCGGTCGT

	AAAACAATTGACTTTAGAAATGAGGAAGCGCAACG

	TCGTCTAACTGAGGCTATCATGTTGCGTGACTTCG

	GCGTTGTGTTAAACATACCATCTAACAGGCTGTGC

	CCGCCTGTGCCGAATCGTATGAACTATGTACTTTG

	GATACAAGATATAGTTTACGCGCACCAGACAATAC

	TGGGAGTGAGTTCTCGTCGTATCAGAGGTCTTGAT

	ATTGGTACTGGTGCTACCGCTATATATCCTATACT

	GGCATGCAAGAAAGAGCAGAGCTGGGAGATGGTTG

	CAACTGAATTGGACGACTACTCCTATGAGTGTGCA

	TGTGATAACGTGTCATCCAACAATATGCAGACTTC

	CATTAAAGTAAAGAAGGCTTCGGTAGATGGGCCCA

	TCCTGTTCCCAGTGGAAAACCAAAATTTCGACTTT

	AGCATGTGCAACCCGCCTTTCTACGGCTCTAAGGA

	GGAGGTGGCGCAATCCGCAGAGTCAAAAGAACTGC

	CGCCCAATGCTGTTTGCACGGGTGCAGAGATCGAG

	ATGATATTTAGTCAAGGAGGAGAAGAGGGTTTCGT

	AGGTAGAATGGTAGAGGAATCAGAGAGGTTGCAAA

	CGAGATGCAAATGGTACACTTCAATGCTTGGTAAG

	ATGTCTAGTGTAAGCACTATAGTTCAGGCTCTGCG

	TGCGAGATCAATTATGAATTATGCTTTGACAGAAT

	TTGTACAAGGACAAACCCGTAGGTGGGCGATAGCT

	TGGTCTTTCTCCGACACTCACTTACCGGATGCCGT

	CAGTAGAATCTCTAGTTAA

	(Psilocybe cubensis (PSID gene))
	SEQ ID NO: 14
	MQVIPACNSAAIRSLCPTPESFRNMGWLSVSDAVY

	SEFIGELATRASNRNYSNEFGLMQPIQEFKAFIES

	DPVVHQEFIDMFEGIQDSPRNYQELCNMFNDIFRK

	APVYGDLGPPVYMIMAKLMNTRAGFSAFTRQRLNL

	HFKKLFDTWGLFLSSKDSRNVLVADQFDDRHCGWL

	NERALSAMVKHYNGRAFDEVFLCDKNAPYYGFNSY

	DDFFNRRFRNRDIDRPVVGGVNNTTLISAACESLS

	YNVSYDVQSLDTLVFKGETYSLKHLLNNDPFTPQF

	EHGSILQGFLNVTAYHRWHAPVNGTIVKIINVPGT

	YFAQAPSTIGDPIPDNDYDPPPYLKSLVYFSNIAA

	RQIMFIEADNKEIGLIFLVFIGMTEISTCEATVSE

	GQHVNRGDDLGMFHFGGSSFALGLRKDCRAEIVEK

	FTEPGTVIRINEVVAALKA

	(Psilocybe cyanescens (PSID gene))
	SEQ ID NO: 15
	MQVLPACQSSALKTLCPSPEAFRKLGWLPTSDEVY

	NEFIDDLTGRTCNEKYSSQVTLLKPIQDFKTFIEN

	DPIVYQEFISMFEGIEQSPTNYHELCNMFNDIFRK

	APLYGDLGPPVYMIMARIMNTQAGFSAFTKESLNF

	HFKKLFDTWGLFLSSKNSRNVLVADQFDDKHYGWF

	SERAKTAMMINYPGRTFEKVFICDEHVPYHGFTSY

	DDFFNRRFRDKDTDRPVVGGVTDTTLIGAACESLS

	YNVSHNVQSLDTLVIKGEAYSLKHLLHNDPFTPQF

	EHGSIIQGFLNVTAYHRWHSPVNGTIVKIVNVPGT

	YFAQAPYTIGSPIPDNDRDPPPYLKSLVYFSNIAA

	RQIMFIEADNKDIGLIFLVFIGMTEISTCEATVCE

	GQHVNRGDDLGMFHFGGSSFALGLRKDSKAKILEK

	FAKPGTVIRINELVASVRK

	(Gymnopilus junonius (PSID gene))
	SEQ ID NO: 16
	MSSPRIVLHRVGGWLPKDQNVLEAWLSKKIAKAKT

	RNRAPKDWAPVIQDFQRLIETDAEIYMGFHQMFEQ

	VPKKTPYDKDPTNEQWQVRNYMHMLDLFDLIITEA

	PDFEQNDLVGFPINAILDWPMGTPGGLTAFINPKV

	NIMFHKMFDVWAVFLSSPASCYVLNTSDSGWFGPA

	ATAAIPNFKETFICDPSLPYLGYTSWDNFFTRLFR

	PGVRPVEFPNNDAIVNSACESTVYNIAPNIKPLDK

	FWIKGEPYSLNHILNNDPYASQFVGGTISQAFLSA

	LNYHRWASPVNGNIVKVVNVPGTYYAESPVTGFGN

	PEGPDPAAPNLSQGFITAVAARALIFIEADNPNIG

	LMCFVGVGMAEVSTCEVTVSVGDVVKKGDEIGMFH

	FGGSTHCLIFRPQTKITFNPDYPVSTAVPLNAAVA

	TVV

	(Psilocybe cubensis (PSIH gene))
	SEQ ID NO: 17
	MIAVLFSFVIAGCIYYIVSRRVRRSRLPPGPPGIP

	IPFIGNMFDMPEESPWLTFLQWGRDYNTDILYVDA

	GGTEMVILNTLETITDLLEKRGSIYSGRLESTMVN

	ELMGWEFDLGFITYGDRWREERRMFAKEFSEKGIK

	QFRHAQVKAAHQLVQQLTKTPDRWAQHIRHQIAAM

	SLDIGYGIDLAEDDPWLEATHLANEGLAIASVPGK

	FWVDSFPSLKYLPAWFPGAVFKRKAKVWREAADHM

	VDMPYETMRKLAPQGLTRPSYASARLQAMDLNGDL

	EHQEHVnCNTAAEVNVGGGDTTVSAMSAFILAMVK

	YPEVQRKVQAELDALTNNGQIPDYDEEDDSLPYLT

	ACIKELFRWNQIAPLAIPHKLMKDDVYRGYLIPKN

	TLVFANTWAVLNDPEVYPDPSVFRPERYLGPDGKP

	DNTVRDPRKAAFGYGRRNCPGIHLAQSTVWIAGAT

	LLSAFNIERPVDQNGKPIDIPADFTTGFFRHPVPF

	QCRFVPRTEQVSQSVSGP

	(Psilocybe cyanescens (PSIH gene))
	SEQ ID NO: 18
	MAPLTTMIPIVLSLLIAGCIYYINARRIKRSRLPP

	GPPGIPIPFIGNMFDMPSESPWLIFLQWGQEYQTD

	IIYVDAGGTDMIILNSLEAITDLLEKRGSLYSGRL

	ESTMVNELMGWEFDFGFIPYGERWREERRMFAKEF

	SEKNIRQFRHAQVKAANQLVRQLTDKPDRWSHHIR

	HQIASMALDIGYGIDLAEDDPWIAASELANEGLAV

	ASVPGSFWVDTFPFLKYLPSWLPGAEFKRNAKMWK

	EGADHMVNMPYETMKKLSAQGLTRPSYASARLQAM

	DPNGDLEHQERVIKNTATQVNVGGGDTTVGAVSAF

	ILAMVKYPEVQRKVQAELDEFTSKGRIPDYDEDND

	SLPYLSACFKELFRWGQIAPLAIAHRLIKDDVYRE

	YTIPKNALVFANNWYGRTVLNDPSEYPNPSEFRPE

	RYLGPDGKPDDTVRDPRKAAFGYGRRVCPGIHLAQ

	STVWIAGVALVSAFNIELPVDKDGKCIDIPAAFTT

	GFFR

	(Gymnopilus junonius (PSIH gene))
	SEQ ID NO: 19
	MMSEMNGMDKLALLTTLLAAGFLYFKNKRRSALPF

	PPGPKKHPLLGNLLDLPKKLEWETYRRWGKEYNSD

	VIHVSAGSVNLIIVNSFEAATDLFDKRSANYSSRP

	QFTMVRELMGWNWLMSALIYGDKWREQRRLFQKHF

	STTNAELYQNTQLEYVRKALQHLLEEPSDFMGITR

	HMAGGVSMSLAYGLNIQKKNDPFVDLAQRAVHSIT

	EASVPGTFWVDVMPWLKYIPEWVPGAGFQKKARVW

	RKLQQDFRQVPYQAALKDMASGKAKPSFASECLET

	IDDNEDAQRQREVIKDTAAIVFAAGADTSLSGIHT

	LFAAMLCYPEVQKKAQEELDRVLGGRRLPEFTDEP

	NMPYISALVKEILRWKPATPIGVPHLASEDDVYNG

	YYIPKRAVVIGNSWAMLHDEETYPDPSTFNPDRFL

	TTNKSTGKLELDPTVRDPALMAFGFGRRMCPGRDV

	ALSVIWLTIASVLATFNITKAIDENGKELEPDVQY

	WSGLIVHPLPFKCTIKPRSKAAEELVKSGADAY

	(Psilocybe cubensis (PSIK gene))
	SEQ ID NO: 20
	MAFDLKTEDGLITYLTKHLSLDVDTSGVKRLSGGF

	VNVTWRIKLNAPYQGHTSIILKHAQPHMSTDEDFK

	IGVERSVYEYQAIKLMMANREVLGGVDGIVSVPEG

	LNYDLENNALIMQDVGKMKTLLDYVTAKPPLATDI

	ARLVGTEIGGFVARLHNIGRERRDDPEFKFFSGNI

	VGRTTSDQLYQTIIPNAAKYGVDDPLLPTVVKDLV

	DDVMHSEETLVMADLWSGNILLQLEEGNPSKLQKI

	YILDWELCKYGPASLDLGYFLGDCYLISRFQDEQV

	GTTMRQAYLQSYARTSKHSINYAKVTAGIAAHIVM

	WTDFMQWGSEEERINFVKKGVAAFHD ARGNNDNG

	EITSTLLKESSTA

	(Psilocybe cyanescens (PSIK gene))
	SEQ ID NO: 21
	MTFDLKTEEGLLSYLTKHLSLDVAPNGVKRLSGGF

	VNVTWRVGLNAPYHGHTSIILKHAQPHLSSDIDFK

	IGVERSAYEYQALKIVSANSSLLGSSDIRVSVPEG

	LHYDVVNNALIMQDVGTMKTLLDYVTAKPPISAEI

	ASLVGSQIGAFIARLHNLGRENKDKDDFKFFSGNI

	VGRTTADQLYQTIIPNAAKYGIDDPILPIVVKELV

	EEVMNSEETLIMADLWSGNILLQFDENSTELTRIW

	LVDWELCKYGPPSLDMGYFLGDCFLVARFQDQLVG

	TSMRQAYLKSYARNVKEPINYAKATAGIGAHLVMW

	TDFMKWGNDEEREEFVKKGVEAFHEANEDNRNGEI

	TSILVKEASRT

	(Psilocybe cyanescens (PSIM gene))
	SEQ ID NO: 22
	MHIRNPYRDGVDYQALAEAFPALKPHVTVNSDNTT

	SIDFAVPEAQRLYTAALLHRDFGLTITLPEDRLCP

	TVPNRLNYVLWVEDILKVTSDALGLPDNRQVKGID

	IGTGASAIYPMLACSRFKTWSMVATEVDQKCIDTA

	RLNVIANNLQERLAIIATSVDGPILVPLLQANSDF

	EYDFTMCNPPFYDGASDMQTSDAAKGFGFGVNAPH

	TGTVLEMATEGGESAFVAQMVRESLNLQTRCRWFT

	SNLGKLKSLYEIVGLLREHQISNYAINEYVQGATR

	RYAIAWSFIDVRLPDHLSRPSNPDLSSLF

	(Psilocybe cubensis (PSIM gene))
	SEQ ID NO: 23
	MHIRNPYRTPIDYQALSEAFPPLKPFVSVNADGTS

	SVDLTIPEAQRAFTAALLHRDFGLTMTIPEDRLCP

	TVPNRLNYVLWIEDIFNYTNKTLGLSDDRPIKGVD

	IGTGASAIYPMLACARFKAWSMVGTEVERKCIDTA

	RLNVVANNLQDRLSILETSIDGPILVPIFEATEEY

	EYEFTMCNPPFYDGAADMQTSDAAKGFGFGVGAPH

	SGTVIEMSTEGGESAFVAQMVRESLKLRTRCRWYT

	SNLGKLKSLKEIVGLLKELEISNYAINEYVQGSTR

	RYAVAWSFTDIQLPEELSRPSNPELSSLF

	(Panaeolus cyanescens (PSIM gene))
	SEQ ID NO: 24
	MHNRNPYRDVIDYQALAEAYPPLKPHVTVNADNTA

	SIDLTIPEVQRQYTAALLHRDFGLTITLPEDRLCP

	TVPNRLNYVLWIEDIFQCTNKALGLSDDRPVKGVD

	IGTGASAIYPMLACARFKQWSMIATEVERKCIDTA

	RLNVLANNLQDRLSILEVSVDGPILVPIFDTFERA

	TSDYEFEFTMCNPPFYDGAADMQTSDAAKGFGFGV

	NAPHSGTVIEMATEGGEAAFVAQMVRESMKLQTRC

	RWFTSNLGKLKSLHEIVALLRESQITNYAINEYVQ

	GTTRRYALAWSFTDIKLTEELYRPSNPELGPLCST

	FV

	(Gymnopilus junonius (PSIM gene))
	SEQ ID NO: 25
	MHSRNPYRSPPDFAALSAAYPPLSPYITTDLSSGR

	KTIDFRNEEAQRRLTEAIMLRDFGVVLNIPSNRLC

	PPVPNRMNYVLWIQDIVYAHQTILGVSSRRIRGLD

	IGTGATAIYPILACKKEQSWEMVATELDDYSYECA

	CDNVSSNNMQTSIKVKKASVDGPILFPVENQNFDF

	SMCNPPFYGSKEEVAQSAESKELPPNAVCTGAEIE

	MIFSQGGEEGFVGRMVEESERLQTRCKWYTSMLGK

	MSSVSTIVQALRARSIMNYALTEFVQGQTRRWAIA

	WSFSDTHLPDAVSRISS

	(Gymnopilus dilepis (PSIM gene))
	SEQ ID NO: 26
	MHIRNPYLTPPDYEALAEAFPALKPYVTVNPDKTT

	TIDFAIPEAQRLYTAALLYRDFGLTITLPPDRLCP

	TVPNRLNYVLWIQDILQITSAALGLPEARQVKGVD

	IGTGAAAIYPILGCSLAKNWSMVGTEVEQKCIDIA

	RQNVISNGLQDRITITANTIDAPILLPLFEGDSNF

	EWEFTMCNPPFYDGAADMETSQDAKGFGFGVNAPH

	TGTVVEMATDGGEAAFVSQMVRESLHLKTRCRWFT

	SNLGKLKSLHEIVGLLREHQITNYAINEYVQGTTR

	RYAIAWSFTDLRLSDHLPRPPNPDLSALF

	(Saccharmyces cerevisiae (ARO1 gene))
	SEQ ID NO: 27
	ATGGTTCAACTAGCCAAGGTTCCAATACTAGGAAA

	CGATATAATACACGTTGGATATAATATACACGATC

	ATCTTGTAGAGACAATTATTAAACACTGTCCTTCT

	TCTACTTACGTCATCTGTAACGATACTAACCTTAG

	CAAGGTACCTTATTACCAGCAACTGGTTCTGGAGT

	TCAAAGCAAGTCTTCCCGAAGGCTCCAGACTACTA

	ACCTACGTGGTCAAACCGGGCGAGACGTCTAAGAG

	TAGGGAGACGAAGGCGCAGTTAGAGGATTATCTTT

	TAGTAGAAGGGTGCACTCGTGATACGGTCATGGTA

	GCCATCGGCGGAGGTGTCATCGGTGACATGATCGG

	TTTCGTAGCCTCCACGTTCATGAGAGGTGTGAGGG

	TAGTACAGGTTCCGACGTCTCTTTTAGCAATGGTA

	GACTCATCCATAGGCGGTAAAACGGCGATCGATAC

	TCCGCTAGGAAAGAACTTCATTGGAGCCTTTTGGC

	AGCCAAAATTTGTTCTTGTGGATATCAAGTGGCTT

	GAAACACTAGCTAAACGTGAATTTATCAACGGCAT

	GGCAGAAGTGATCAAGACAGCGTGCATCTGGAACG

	CTGATGAATTTACTCGTCTCGAATCCAACGCGTCA

	CTGTTCCTAAACGTAGTAAATGGTGCGAAAAATGT

	AAAGGTGACTAACCAGCTGACGAACGAGATAGATG

	AGATCAGCAACACGGATATTGAAGCCATGTTGGAC

	CATACTTATAAACTGGTATTAGAGAGTATTAAGGT

	TAAAGCGGAGGTGGTAAGCAGCGATGAAAGGGAGA

	GCAGTCTTAGGAACCTTTTAAACTTCGGGCATAGC

	ATAGGTCACGCGTATGAAGCCATACTGACACCCCA

	GGCTTTACATGGAGAGTGCGTATCCATCGGCATGG

	TAAAAGAAGCAGAACTATCAAGGTATTTTGGGATA

	CTTTCTCCGACCCAGGTGGCGCGTCTAAGCAAAAT

	TCTAGTTGCGTACGGATTGCCCGTTAGCCCCGATG

	AGAAATGGTTTAAAGAGCTTACACTTCATAAGAAG

	ACACCCTTGGACATACTGCTAAAGAAGATGAGCAT

	CGACAAGAAAAATGAAGGAAGCAAGAAGAAGGTCG

	TAATCCTAGAGTCTATCGGCAAATGTTACGGAGAC

	TCAGCTCAGTTTGTTTCAGACGAAGACTTACGTTT

	TATATTGACAGATGAAACACTAGTATATCCTTTTA

	AGGATATTCCCGCTGATCAGCAGAAAGTCGTGATT

	CCACCCGGAAGTAAATCAATAAGCAATCGTGCTTT

	AATCTTAGCAGCTCTGGGGGAGGGACAGTGCAAGA

	TCAAGAACCTATTACACTCCGACGACACCAAACAT

	ATGCTGACCGCAGTCCACGAGTTAAAAGGTGCTAC

	CATCAGTTGGGAGGATAACGGAGAAACAGTGGTCG

	TAGAGGGCCATGGCGGGAGCACTCTATCGGCTTGT

	GCTGATCCCTTATACTTAGGCAACGCGGGGACGGC

	GAGTAGATTCTTAACATCACTGGCGGCACTAGTGA

	ACAGTACATCCTCCCAAAAGTATATCGTACTAACA

	GGCAACGCAAGGATGCAGCAACGTCCGATAGCGCC

	CCTTGTTGACAGCTTACGTGCTAACGGGACAAAGA

	TCGAGTACTTGAACAACGAAGGTTCTTTGCCGATC

	AAAGTGTACACTGATTCTGTATTTAAAGGCGGCCG

	TATTGAGTTGGCTGCGACAGTTAGTTCCCAATACG

	TGAGCAGTATCCTGATGTGTGCGCCTTACGCAGAA

	GAGCCCGTGACTTTAGCTTTGGTAGGTGGGAAACC

	GATCAGTAAACTATACGTTGATATGACAATTAAGA

	TGATGGAAAAGTTCGGCATCAATGTGGAGACCTCA

	ACCACGGAACCCTACACATACTACATTCCGAAGGG

	GCATTACATTAATCCAAGTGAGTACGTAATCGAGA

	GCGACGCTTCATCCGCTACCTATCCGTTAGCATTC

	GCCGCAATGACCGGTACCACCGTAACAGTCCCCAA

	CATCGGCTTTGAATCTCTGCAGGGCGACGCTAGAT

	TCGCAAGAGACGTCCTAAAGCCGATGGGGTGTAAA

	ATCACCCAAACGGCTACGTCTACAACCGTCAGTGG

	ACCACCCGTCGGTACGCTAAAGCCATTAAAACACG

	TTGATATGGAACCAATGACAGACGCCTTCTTAACC

	GCATGCGTTGTAGCCGCAATCAGTCATGACTCCGA

	CCCCAATTCAGCGAACACTACTACTATCGAGGGGA

	TCGCAAACCAAAGGGTTAAAGAATGCAACAGAATC

	TTAGCGATGGCTACCGAGCTGGCAAAGTTTGGAGT

	AAAGACAACAGAACTTCCCGATGGCATACAGGTCC

	ATGGGCTAAATTCCATCAAGGACCTTAAAGTCCCA

	TCTGACAGCTCAGGACCCGTCGGAGTCTGTACTTA

	TGATGACCATAGGGTTGCCATGTCATTTTCCCTTT

	TGGCTGGCATGGTAAACAGTCAGAATGAGAGAGAT

	GAAGTGGCAAACCCAGTTAGGATCTTAGAGAGGCA

	CTGCACCGGAAAGACGTGGCCAGGCTGGTGGGACG

	TTCTGCACAGCGAACTTGGAGCGAAGCTGGATGGT

	GCCGAGCCGCTAGAATGCACATCCAAAAAGAACTC

	TAAGAAGAGCGTAGTCATAATAGGCATGAGAGCTG

	CGGGCAAAACTACTATCTCTAAGTGGTGCGCAAGT

	GCGCTGGGTTACAAGTTGGTAGATTTAGATGAATT

	GTTCGAGCAGCAGCATAATAACCAATCAGTAAAAC

	AATTTGTAGTCGAGAATGGTTGGGAGAAATTCAGA

	GAGGAAGAGACCAGGATATTCAAGGAGGTTATTCA

	AAATTACGGCGACGACGGGTATGTCTTTAGCACTG

	GGGGAGGGATCGTCGAATCCGCGGAGAGCAGGAAA

	GCACTAAAGGACTTCGCCAGTTCCGGTGGGTATGT

	GCTTCACTTACATCGTGATATAGAGGAGACGATAG

	TCTTCCTACAAAGTGATCCATCCAGGCCGGCGTAT

	GTTGAGGAGATTAGGGAGGTCTGGAACCGTAGAGA

	AGGCTGGTATAAAGAATGTAGTAATTTTAGCTTTT

	TCGCACCTCACTGTAGCGCAGAGGCGGAGTTTCAA

	GCACTTAGACGTTCATTCAGTAAGTATATAGCTAC

	GATCACGGGGGTCCGTGAAATAGAGATTCCTAGTG

	GGAGGAGTGCGTTTGTATGCTTAACTTTTGACGAT

	CTAACTGAGCAAACGGAGAATCTGACGCCTATATG

	CTACGGGTGTGAAGCCGTAGAGGTGCGTGTTGATC

	ATCTTGCCAATTATTCCGCAGACTTCGTTAGCAAG

	CAATTAAGCATACTGAGAAAAGCGACCGACAGTAT

	ACCCATTATCTTCACCGTCCGTACTATGAAACAAG

	GCGGTAATTTTCCCGATGAAGAGTTCAAGACATTG

	CGTGAGTTGTACGACATAGCTCTTAAAAACGGAGT

	GGAGTTCCTTGATTTGGAACTTACTCTGCCTACAG

	ATATACAGTACGAAGTCATCAACAAGAGAGGTAAT

	ACGAAGATCATTGGGTCTCATCATGACTTCCAGGG

	TTTGTACAGCTGGGACGATGCTGAATGGGAAAACA

	GATTCAATCAGGCACTGACTCTTGACGTAGATGTG

	GTGAAATTTGTGGGTACCGCGGTGAATTTCGAGGA

	CAACTTACGTTTGGAACATTTTCGTGACACGCACA

	AAAATAAACCACTAATAGCAGTTAACATGACGTCT

	AAGGGCTCAATCAGTAGGGTACTAAATAATGTATT

	GACTCCGGTTACTTCAGACCTTTTACCGAACAGCG

	CAGCGCCTGGTCAATTGACGGTTGCACAGATTAAT

	AAAATGTATACATCTATGGGAGGAATTGAGCCTAA

	AGAGCTATTTGTGGTGGGGAAGCCAATCGGCCACT

	CAAGATCACCTATACTACACAATACTGGGTATGAG

	ATTTTGGGTCTACCTCACAAATTCGATAAATTTGA

	GACGGAAAGCGCACAATTAGTGAAGGAGAAATTGT

	TAGACGGGAACAAGAATTTCGGTGGTGCAGCGGTG

	ACCATCCCTTTAAAGCTAGACATAATGCAGTACAT

	GGATGAACTTACGGACGCTGCGAAGGTGATTGGGG

	CGGTAAACACAGTAATCCCTTTGGGTAACAAGAAA

	TTCAAGGGTGATAATACGGACTGGTTAGGGATAAG

	GAACGCACTTATAAATAATGGTGTGCCCGAGTACG

	TGGGGCATACTGCCGGACTTGTAATAGGTGCTGGT

	GGTACCAGTAGGGCGGCACTGTACGCTTTGCATAG

	CTTAGGTTGCAAGAAGATCTTTATCATCAATAGAA

	CAACTAGTAAACTGAAGCCACTGATAGAATCACTA

	CCCTCCGAGTTTAACATCATTGGAATAGAGTCTAC

	GAAATCCATCGAGGAGATTAAAGAACACGTCGGAG

	TCGCTGTTAGCTGCGTGCCTGCCGATAAGCCCTTA

	GATGACGAGCTACTGAGTAAGTTAGAACGTTTCCT

	TGTCAAGGGTGCACATGCGGCTTTCGTCCCAACAC

	TGCTAGAGGCTGCCTATAAACCCAGCGTAACACCT

	GTTATGACCATAAGTCAGGACAAGTATCAATGGCA

	CGTGGTGCCGGGTTCCCAGATGCTGGTCCATCAAG

	GTGTTGCACAATTTGAAAAATGGACTGGTTTCAAG

	GGGCCCTTCAAAGCCATATTTGACGCCGTGACTAA

	AGAGTAA

	(Saccharomyces cerevisiae (ARO2 gene))
	SEQ ID NO: 28
	ATGTCCACATTCGGTAAACTTTTCCGTGTCACTAC

	ATACGGCGAGTCACACTGCAAATCTGTGGGGTGCA

	TAGTAGACGGCGTTCCGCCGGGCATGAGTTTAACC

	GAAGCGGACATTCAACCTCAGCTTACCCGTAGGAG

	GCCCGGTCAGAGCAAGTTATCCACCCCGAGGGACG

	AAAAGGACCGTGTAGAGATCCAAAGCGGAACGGAA

	TTTGGGAAGACACTTGGTACGCCTATCGCTATGAT

	GATTAAAAACGAGGATCAACGTCCGCACGATTACT

	CCGACATGGACAAGTTCCCTAGGCCGAGTCACGCC

	GATTTTACGTACTCAGAGAAATACGGAATAAAAGC

	CTCCAGCGGTGGGGGCCGTGCTTCCGCGAGAGAAA

	CCATTGGAAGAGTAGCATCCGGTGCAATAGCAGAG

	AAGTTCCTAGCACAGAACTCAAATGTTGAAATTGT

	CGCTTTCGTCACGCAAATAGGTGAGATCAAGATGA

	ACCGTGACAGTTTCGACCCAGAATTTCAACACCTT

	CTAAATACAATTACGAGGGAGAAGGTAGATAGCAT

	GGGTCCAATAAGATGCCCCGACGCTTCCGTCGCGG

	GATTGATGGTGAAGGAAATTGAAAAATATCGTGGG

	AACAAGGATTCTATTGGGGGTGTAGTAACTTGCGT

	AGTCAGAAATCTACCTACAGGGTTGGGTGAACCGT

	GTTTTGACAAACTGGAGGCGATGCTGGCACATGCC

	ATGTTATCCATACCAGCAAGTAAAGGATTTGAAAT

	AGGATCTGGCTTCCAGGGTGTAAGCGTACCAGGAA

	GCAAACACAATGATCCCTTTTACTTTGAAAAAGAG

	ACTAACCGTCTTCGTACAAAGACAAACAACTCCGG

	TGGGGTGCAAGGGGGCATCTCTAATGGTGAGAACA

	TTTACTTTTCCGTACCATTTAAGAGCGTGGCTACA

	ATAAGCCAAGAGCAAAAGACCGCAACTTACGATGG

	AGAAGAAGGAATCCTCGCAGCTAAGGGTAGGCACG

	ATCCTGCGGTCACACCGCGTGCAATTCCCATAGTG

	GAAGCTATGACCGCCCTAGTACTAGCAGATGCGTT

	ACTAATACAGAAAGCCAGGGATTTTTCTAGGTCAG

	TCGTACATTAA

	(Saccharomyces cerevisiae (ARO3 gene))
	SEQ ID NO: 29
	ATGTTCATCAAGAATGACCATGCTGGTGATAGAAA

	GAGACTAGAGGACTGGCGTATAAAGGGTTATGACC

	CTCTAACTCCGCCTGATTTGCTACAGCACGAGTTT

	CCTATATCAGCAAAAGGGGAAGAAAATATCATCAA

	GGCTCGTGATAGTGTATGTGATATACTGAACGGAA

	AGGATGACAGACTTGTGATAGTAATTGGACCCTGT

	TCTCTGCATGATCCGAAGGCGGCCTACGACTATGC

	CGACAGATTAGCCAAAATATCCGAAAAGCTGTCAA

	AAGATCTTTTAATTATCATGCGTGCATACCTAGAG

	AAGCCTCGTACAACCGTTGGATGGAAAGGGTTGAT

	AAACGACCCGGATATGAACAATAGTTTTCAGATTA

	ATAAAGGCCTTCGTATAAGCCGTGAGATGTTTATA

	AAACTAGTTGAGAAATTACCTATTGCAGGAGAAAT

	GCTTGACACGATTTCCCCTCAGTTCTTATCTGACT

	GTTTCTCACTAGGTGCAATTGGTGCTAGGACTACC

	GAGTCACAGTTACATCGTGAACTGGCCAGCGGTCT

	GTCTTTCCCCATTGGCTTTAAAAATGGTACCGATG

	GTGGCCTTCAAGTAGCAATTGATGCTATGAGAGCT

	GCGGCCCACGAACACTACTTTTTGTCTGTGACCAA

	ACCTGGCGTAACAGCGATTGTGGGAACTGAAGGGA

	ACAAGGACACCTTCCTAATCCTGAGAGGGGGCAAG

	AACGGGACTAATTTTGACAAGGAGTCAGTTCAAAA

	CACTAAGAAGCAATTGGAGAAGGCGGGCCTTACTG

	ACGATTCTCAGAAGAGAATCATGATAGACTGCAGC

	CATGGCAACTCAAATAAAGATTTCAAAAATCAACC

	CAAAGTCGCCAAGTGTATCTACGATCAACTAACCG

	AAGGAGAAAATAGTTTATGCGGGGTGATGATAGAG

	AGTAATATAAACGAAGGAAGACAGGATATTCCTAA

	GGAAGGCGGAAGAGAGGGTCTGAAGTACGGGTGTT

	CTGTGACAGACGCTTGCATAGGATGGGAGAGCACG

	GAACAGGTTTTGGAGCTGCTGGCAGAAGGGGTGCG

	TAATAGAAGGAAAGCCTTAAAGAAGTAA

	(Saccharomyces cerevisiae (ARO4 K2229L gene))
	SEQ ID NO: 30
	ATGAGCGAATCTCCGATGTTCGCCGCAAACGGCAT

	GCCTAAGGTAAATCAAGGGGCCGAGGAGGACGTGA

	GAATATTAGGTTATGACCCGCTTGCCAGTCCTGCA

	TTGCTTCAGGTACAGATTCCAGCAACGCCAACGTC

	CTTAGAAACAGCAAAAAGGGGACGTCGTGAAGCTA

	TAGACATCATCACTGGCAAGGACGACCGTGTCCTA

	GTAATAGTTGGTCCGTGCTCTATCCATGACCTTGA

	GGCTGCACAGGAGTATGCACTAAGGTTGAAGAAAT

	TGTCTGATGAACTGAAAGGTGATCTTAGTATAATC

	ATGCGTGCATATTTAGAGAAACCGCGTACGACGGT

	AGGCTGGAAAGGGCTAATTAACGATCCGGATGTGA

	ATAATACCTTTAACATCAACAAGGGTCTACAGAGT

	GCGCGTCAGTTATTCGTGAACTTAACAAATATCGG

	ACTGCCGATAGGCTCCGAGATGCTGGACACGATAT

	CTCCCCAGTATTTGGCTGACCTTGTTTCTTTTGGA

	GCTATAGGTGCAAGGACTACTGAGAGTCAGTTACA

	TAGAGAGTTGGCATCAGGACTTAGCTTCCCTGTAG

	GATTTAAGAACGGTACAGACGGCACTCTTAATGTC

	GCGGTCGATGCCTGCCAGGCAGCCGCCCATTCACA

	TCATTTTATGGGAGTGACATTACACGGGGTGGCCG

	CTATCACAACGACTAAAGGGAATGAGCACTGTTTT

	GTTATCCTTAGAGGAGGAAAGAAAGGTACGAATTA

	TGATGCGAAAAGTGTAGCAGAGGCCAAAGCGCAAC

	TTCCTGCCGGTTCAAACGGACTTATGATTGACTAT

	TCCCATGGAAACTCAAATAAGGACTTTAGGAATCA

	GCCAAAAGTTAACGATGTGGTATGCGAACAGATCG

	CGAACGGTGAAAATGCGATTACGGGTGTTATGATC

	GAGTCAAATATAAATGAAGGTAACCAAGGTATCCC

	GGCAGAGGGCAAAGCGGGCCTGAAGTACGGTGTAT

	CTATTACGGATGCCTGTATAGGTTGGGAGACAACC

	GAAGACGTCCTAAGGAAACTTGCCGCCGCGGTTAG

	ACAGAGACGTGAAGTCAATAAGAAGTAA

	(Escherichia coli (AROL gene))
	SEQ ID NO: 31
	ATGACCCAGCCATTATTTCTGATCGGTCCTCGTGG

	GTGCGGGAAAACGACGGTTGGCATGGCCTTAGCTG

	ACAGTTTGAATCGTAGATTCGTGGACACCGACCAG

	TGGCTACAGTCTCAGCTTAACATGACGGTGGCCGA

	AATTGTAGAACGTGAAGAATGGGCTGGTTTTCGTG

	CAAGAGAAACAGCCGCATTGGAAGCTGTGACGGCG

	CCTTCAACGGTGATAGCTACGGGAGGTGGTATTAT

	TTTGACCGAATTTAATAGGCACTTCATGCAGAATA

	ATGGCATAGTGGTTTACCTATGCGCTCCTGTGTCT

	GTCTTGGTAAACCGTTTGCAAGCCGCACCAGAAGA

	AGACTTGCGTCCAACCCTGACGGGGAAGCCACTGT

	CTGAGGAAGTGCAAGAGGTACTGGAGGAAAGGGAC

	GCTCTATACCGTGAGGTGGCTCACATCATAATTGA

	CGCTACGAATGAGCCATCACAGGTAATTTCTGAGA

	TCCGTTCAGCGTTGGCCCAAACCATCAATTGTTAA


	(Saccharomyces cerevisiae (TRP1 gene))
	SEQ ID NO: 32
	ATGTCAGTGATTAACTTTACAGGCTCCTCAGGTCC

	CTTGGTCAAGGTCTGCGGCTTGCAATCAACAGAGG

	CCGCTGAATGCGCCCTAGACTCAGATGCAGACCTT

	TTAGGCATCATCTGTGTCCCCAACAGAAAGCGTAC

	TATTGATCCTGTTATTGCGCGTAAGATCAGTTCTT

	TGGTCAAGGCGTATAAGAACTCCTCAGGAACCCCC

	AAGTATCTGGTAGGGGTATTCAGGAATCAACCTAA

	AGAAGACGTCTTGGCCCTAGTTAATGACTACGGCA

	TAGACATAGTCCAGTTGCACGGAGACGAAAGCTGG

	CAAGAATATCAGGAATTTTTGGGGCTGCCGGTTAT

	AAAAAGGCTGGTTTTCCCTAAGGACTGTAACATAC

	TGTTATCAGCCGCATCACAGAAGCCGCATTCCTTT

	ATACCTCTTTTCGACTCCGAGGCCGGAGGCACTGG

	TGAATTACTGGACTGGAACAGCATTTCAGATTGGG

	TAGGGAGGCAGGAGAGCCCAGAATCTCTTCATTTT

	ATGTTGGCAGGGGGCCTTACGCCGGAAAATGTTGG

	AGATGCATTGAGGTTGAACGGAGTTATAGGTGTGG

	ATGTCAGTGGTGGGGTTGAAACGAATGGTGTTAAA

	GACAGCAACAAAATAGCAAATTTTGTCAAGAATGC

	CAAAAAGTAA

	(Saccharomyces cerevisiae (TRP2 S76L gene))
	SEQ ID NO: 33
	ATGACGGCGAGCATTAAAATTCAGCCAGACATTGA

	CAGTTTAAAGCAGTTGCAGCAACAGAATGACGACT

	CTTCCATTAACATGTATCCCGTGTATGCGTATCTG

	CCTTCTTTGGATTTGACACCTCACGTTGCTTACTT

	AAAGTTAGCTCAACTTAATAATCCAGATAGAAAGG

	AGTCTTTCTTACTTGAAAGTGCTAAGACCAATAAT

	GAGCTGGACAGATATCTTTTCATAGGGATCAGTCC

	AAGGAAGACCATTAAGACCGGGCCCACTGAAGGCA

	TTGAGACTGACCCATTAGAAATCCTTGAAAAAGAA

	ATGTCTACTTTCAAAGTCGCCGAAAACGTCCCAGG

	CCTTCCCAAATTAAGCGGCGGGGCGATAGGTTACA

	TATCATACGACTGTGTACGTTACTTCGAACCCAAG

	ACTAGGCGTCCCTTGAAAGATGTGCTTAGGTTACC

	AGAGGCGTACTTGATGCTTTGTGACACGATAATCG

	CATTTGACAATGTCTTCCAAAGGTTTCAAATTATT

	CACAATATTAACACAAACGAAACGTCTTTGGAGGA

	AGGATACCAGGCGGCTGCGCAGATAATCACGGATA

	TTGTATCTAAGTTGACAGACGACAGCTCCCCCATT

	CCGTACCCGGAGCAACCCCCTATCAAACTAAACCA

	AACCTTTGAATCCAACGTAGGCAAAGAGGGGTATG

	AAAATCACGTCTCCACTCTCAAAAAGCACATAAAG

	AAAGGTGACATAATCCAAGGTGTGCCCAGCCAGAG

	AGTGGCGAGGCCTACATCTTTACATCCATTCAACA

	TATATAGGCATCTTAGAACCGTGAACCCATCACCT

	TATCTATTTTACATAGACTGCCTAGATTTCCAGAT

	AATAGGGGCTAGTCCCGAATTGCTGTGTAAATCAG

	ATTCAAAGAATCGTGTTATTACACACCCCATAGCT

	GGCACAGTCAAGAGGGGTGCTACCACTGAGGAAGA

	TGACGCTCTGGCAGATCAGCTACGTGGTTCTTTGA

	AAGATAGGGCTGAGCATGTTATGCTGGTTGACTTA

	GCAAGAAACGACATCAATCGTATATGCGATCCCCT

	AACGACTTCCGTTGACAAACTTTTGACCATTCAGA

	AGTTCAGCCACGTACAGCACTTAGTCTCTCAGGTC

	TCTGGCGTCCTAAGGCCTGAGAAAACTCGTTTCGA

	TGCATTCAGAAGCATATTTCCCGCGGGTACAGTGA

	GTGGGGCCCCAAAGGTGCGTGCAATGGAGCTTATA

	GCCGAGCTAGAAGGCGAGCGTAGGGGAGTGTACGC

	AGGGGCCGTAGGCCATTGGTCTTATGACGGCAAGA

	CCATGGATAATTGTATTGCACTAAGGACCATGGTC

	TATAAAGATGGGATTGCATACTTGCAGGCAGGAGG

	TGGGATTGTCTATGACAGCGATGAGTACGATGAGT

	ATGTAGAAACAATGAATAAAATGATGGCGAATCAT

	TCCACGATAGTGCAGGCGGAGGAGTTATGGGCGGA

	TATTGTGGGTAGTGCATAA

	(Saccharomyces cerevisiae (TRP3 gene))
	SEQ ID NO: 34
	ATGTCTGTCCACGCAGCCACCAACCCGATAAATAA

	GCATGTCGTTCTGATTGATAATTACGACTCCTTCA

	CGTGGAATGTTTATGAGTATCTTTGCCAGGAGGGA

	GCGAAGGTTAGCGTTTACCGTAATGACGCTATCAC

	GGTCCCAGAAATTGCAGCACTGAATCCCGATACCC

	TTCTGATATCACCAGGCCCGGGCCATCCCAAGACA

	GATTCTGGTATTAGCAGAGATTGCATCAGATACTT

	CACTGGAAAAATTCCAGTTTTTGGGATATGTATGG

	GGCAGCAATGCATGTTTGACGTGTTTGGCGGGGAA

	GTGGCTTATGCGGGTGAAATAGTGCACGGAAAGAC

	TAGTCCCATATCCCATGATAACTGCGGTATCTTTA

	AGAATGTCCCCCAGGGTATTGCAGTTACAAGATAT

	CATAGCTTGGCTGGCACTGAAAGTAGTCTGCCTAG

	CTGCCTAAAGGTGACTGCCTCTACTGAAAACGGGA

	TAATCATGGGGGTAAGGCACAAGAAGTACACCGTC

	GAGGGGGTGCAATTCCACCCAGAGAGTATTTTAAC

	CGAAGAAGGACATCTAATGATCCGTAATATTCTTA

	ATGTTTCTGGCGGAACGTGGGAGGAAAATAAATCA

	AGCCCATCCAATTCCATCCTAGATAGGATATACGC

	CAGGCGTAAAATTGACGTAAACGAACAGTCAAAGA

	TTCCCGGTTTCACCTTTCAGGACTTACAATCTAAC

	TATGATCTTGGCCTTGCCCCGCCTCTGCAAGATTT

	TTATACCGTGCTGAGCAGTAGTCATAAGAGGGCTG

	TGGTCCTAGCGGAGGTGAAGCGTGCCTCCCCTAGC

	AAAGGTCCAATCTGCCTGAAGGCCGTTGCTGCTGA

	ACAAGCCCTTAAATATGCTGAGGCTGGGGCGAGTG

	CAATTAGCGTTCTAACAGAACCCCACTGGTTCCAC

	GGGAGCCTTCAAGACCTTGTGAATGTAAGAAAGAT

	CTTGGATCTAAAATTTCCGCCAAAAGAGAGACCCT

	GCGTGCTTAGGAAAGAGTTTATATTTTCCAAATAC

	CAAATATTGGAGGCACGTCTAGCTGGTGCAGATAC

	TGTCCTTTTGATTGTAAAGATGTTGTCCCAACCAT

	TACTGAAAGAGCTATATAGTTACTCAAAGGATTTA

	AACATGGAGCCGTTAGTGGAAGTAAATAGCAAGGA

	GGAGCTACAACGTGCCCTGGAAATTGGTGCCAAGG

	TTGTTGGAGTTAACAATCGTGACTTGCATTCCTTC

	AACGTAGACTTGAATACAACAAGTAATTTGGTCGA

	ATCTATCCCAAAAGATGTGCTGTTGATTGCACTTT

	CCGGTATCACAACACGTGATGACGCCGAAAAGTAT

	AAAAAGGAGGGGGTGCACGGGTTTTTGGTGGGTGA

	GGCGTTAATGAAATCTACAGATGTAAAGAAGTTTA

	TTCATGAGCTGTGCGAATAA

	(Saccharomyces cerevisiae (TRP4 gene))
	SEQ ID NO: 35
	ATGAGCGAAGCTACTCTATTAAGTTATACCAAAAA

	GCTACTAGCAAGCCCACCTCAGCTTAGTTCCACCG

	ACCTACACGATGCACTACTTGTCATCCTAAGTCTA

	CTTCAGAAGTGCGACACCAATTCTGATGAGTCCTT

	GTCTATTTATACGAAGGTGTCTTCCTTTTTAACAG

	CCCTAAGGGTGACTAAGTTAGATCATAAGGCGGAA

	TATATTGCCGAGGCTGCAAAAGCAGTTTTGCGTCA

	CTCAGATCTGGTCGATCTACCTTTACCTAAAAAGG

	ATGAGCTGCATCCTGAAGATGGTCCTGTTATCTTG

	GACATTGTGGGTACTGGGGGTGATGGACAGAATAC

	CTTTAACGTGTCAACGTCAGCCGCTATTGTGGCCT

	CAGGTATTCAGGGACTGAAGATTTGCAAACACGGA

	GGTAAAGCATCTACCTCAAACAGCGGAGCTGGAGA

	TCTGATTGGGACATTGGGATGCGATATGTTCAAAG

	TGAATAGTAGCACAGTCCCCAAATTGTGGCCAGAC

	AATACATTTATGTTCTTATTGGCTCCATTCTTTCA

	TCATGGGATGGGTCATGTAAGCAAGATTCGTAAGT

	TTCTTGGAATACCTACGGTATTTAACGTATTGGGG

	CCGCTGTTACACCCCGTATCCCATGTGAATAAGAG

	GATACTTGGAGTGTATTCAAAAGAGTTGGCGCCAG

	AATATGCGAAGGCAGCAGCCTTGGTCTATCCAGGG

	TCAGAAACGTTTATTGTGTGGGGCCATGTTGGGCT

	TGACGAGGTGAGCCCCATAGGAAAGACTACCGTGT

	GGCACATCGATCCGACAAGCTCAGAACTAAAGTTG

	AAGACCTTCCAGCTGGAGCCATCTATGTTCGGTCT

	GGAGGAGCACGAGCTGAGTAAATGCGCCTCATATG

	GACCTAAGGAGAATGCTCGTATATTAAAGGAGGAA

	GTCCTTTCCGGCAAATACCACCTAGGCGACAATAA

	TCCAATATATGATTACATTCTGATGAATACTGCAG

	TATTATACTGCCTGTCCCAAGGGCACCAAAACTGG

	AAGGAAGGTATTATCAAAGCCGAGGAGTCAATTCA

	CAGCGGGAATGCCTTGAGATCGCTAGAACATTTCA

	TTGATTCAGTATCTTCCCTTTAA

	(Saccharomyces cerevisiae (TAT2 gene))
	SEQ ID NO: 36
	ATGACCGAAGATTTCATCAGTAGCGTCAAAAGGTC

	AAATGAAGAGCTTAAAGAGAGAAAATCTAATTTTG

	GGTTTGTAGAGTACAAGTCAAAACAACTTACCTCC

	AGTAGCTCACACAACTCCAACTCTTCACACCATGA

	TGACGACAACCAGCACGGTAAAAGAAACATCTTTC

	AGCGTTGTGTGGATTCTTTTAAATCCCCTCTGGAT

	GGGTCTTTCGACACCTCCAATCTGAAAAGAACACT

	GAAACCTCGTCATTTAATAATGATCGCAATAGGAG

	GTAGTATAGGTACTGGTCTTTTCGTGGGTTCAGGG

	AAGGCTATAGCGGAAGGCGGACCACTTGGCGTTGT

	GATCGGATGGGCCATTGCGGGTAGCCAAATAATAG

	GTACTATACATGGGTTAGGAGAGATCACGGTAAGA

	TTTCCAGTAGTCGGTGCGTTTGCCAACTACGGCAC

	CCGTTTCTTGGACCCGAGCATTAGTTTTGTAGTCT

	CCACTATATACGTGCTACAGTGGTTCTTTGTCCTA

	CCCCTAGAGATTATTGCTGCGGCGATGACCGTGCA

	ATACTGGAACAGTTCTATCGATCCGGTAATATGGG

	TCGCAATTTTCTATGCCGTCATCGTCTCAATCAAT

	TTGTTTGGAGTTAGGGGTTTCGGAGAAGCTGAATT

	CGCCTTCTCAACTATTAAGGCAATCACTGTCTGTG

	GCTTCATAATCTTATGTGTAGTCTTGATCTGCGGC

	GGAGGACCCGATCACGAATTCATTGGTGCTAAATA

	CTGGCATGATCCTGGCTGCCTGGCAAACGGGTTTC

	CTGGAGTCTTGAGTGTCCTTGTCGTTGCGTCATAC

	AGCCTAGGAGGCATAGAAATGACTTGCTTAGCCTC

	TGGGGAAACGGACCCAAAGGGACTTCCCTCAGCTA

	TAAAACAGGTTTTCTGGCGTATTTTGTTTTTCTTC

	TTAATTTCTTTAACTCTAGTGGGATTTTTAGTTCC

	TTACACCAACCAAAATCTACTAGGTGGCTCCTCTG

	TCGATAATAGTCCCTTCGTTATCGCGATTAAGCTA

	CACCATATCAAAGCTCTTCCGTCTATTGTTAACGC

	AGTTATCCTTATTTCCGTGCTATCCGTGGGTAACA

	GTTGCATCTTTGCCAGCTCCAGAACTCTGTGTAGC

	ATGGCACATCAAGGACTGATACCGTGGTGGTTCGG

	CTATATTGACAGAGCTGGCAGACCCCTGGTTGGGA

	TTATGGCCAATTCTCTTTTCGGCTTATTGGCGTTC

	CTTGTTAAATCTGGCTCCATGAGTGAGGTGTTTAA

	TTGGCTGATGGCTATAGCCGGACTGGCGACATGTA

	TTGTGTGGTTATCTATAAATCTTTCCCATATAAGA

	TTCCGTCTTGCAATGAAGGCCCAAGGAAAGTCCCT

	GGATGAACTTGAATTCGTAAGCGCGGTTGGTATAT

	GGGGATCTGCTTATTCCGCACTTATCAATTGCTTA

	ATACTTATTGCTCAATTTTATTGCTCTTTATGGCC

	AATCGGGGGTTGGACATCCGGAAAAGAGAGGGCAA

	AGATTTTCTTTCAGAATTATCTTTGCGCCCTGATT

	ATGTTATTTATATTCATCGTCCATAAGATCTATTA

	TAAATGTCAAACGGGAAAGTGGTGGGGTGTTAAAG

	CTCTGAAGGACATCGACCTAGAGACCGACAGGAAG

	GACATAGACATCGAAATAGTTAAACAAGAAATCGC

	TGAAAAGAAGATGTATTTGGACTCCAGACCTTGGT

	ACGTGAGGCAGTTTCATTTTTGGTGCTAA

Claims

1-21. (canceled)

22. A recombinant host organism comprising:

a plurality of cells transfected by a non-natural gene expressed in the recombinant host organism;

wherein the recombinant host organism is a fungal species selected from the group consisting of Schizosaccharomyces cerevisiae, Schizosaccharomyces japonicus, Schizosaccharomyces pombe, Schizosaccharomyces cryophilus, Saccharomyces cerevisiae, Kluyveromyces lactis, Kluyveromyces dobzhanskii, and Yarrowia lipolytica;

wherein the non-natural gene is selected from a group consisting of PsiD, PsiH, PsiK, and PsiM, wherein:

PsiD encodes an L-tryptophan decarboxylase comprising an amino acid sequence having at least 90% identity to any one of amino acid sequences set forth in SEQ ID NO:14-SEQ ID NO:16;

PsiH encodes a tryptamine 4-monooxygenase comprising an amino acid sequence having at least 90% identity to any one of amino acid sequences set forth in SEQ ID NO:17-SEQ ID NO:19;

PsiK encodes a 4-hydroxytryptamine kinase comprising an amino acid sequence having at least 90% identity to any one of amino acid sequences set forth in SEQ ID NO:20 or SEQ ID NO:21; and

PsiM encodes a methyl transferase comprising an amino acid sequence having at least 90% identity to any one of amino acid sequences set forth in SEQ ID NO:22-SEQ ID NO:26.

23. The recombinant host organism of claim 22, wherein

the L-tryptophan decarboxylase comprises any one of amino acid sequences set forth in SEQ ID NO:14-SEQ ID NO:16;

the tryptamine 4-monooxygenase comprises any one of amino acid sequences set forth in SEQ ID NO:17-SEQ ID NO:19;

the 4-hydroxytryptamine kinase comprises any one of amino acid sequences set forth in SEQ ID NO:20 or SEQ ID NO:21; and

the methyl transferase comprises any one of amino acid sequences set forth in SEQ ID NO:22-SEQ ID NO:26.

24. The recombinant host organism of claim 22, comprising PsiD, PsiH, PsiK and PsiM, wherein the organism synthesizes psilocybin.

25. The recombinant host organism of claim 22, further comprising at least one non-natural gene selected from the group consisting of: SEQ ID NO:27-SEQ ID NO:35.

26. The recombinant host organism of claim 22, further comprising a recombinant transporter protein that is codon optimized for expression in the recombinant host organism.

27. The recombinant host organism of claim 26, wherein the recombinant transporter protein comprises SEQ ID NO:36.

28. The recombinant host organism of claim 22, growing in a medium comprising glucose, galactose, sucrose, fructose, molasses, or any combination thereof.

29. A method, the method comprising:

transfecting a plurality of cells in a recombinant host organism with a set of genes comprising PsiD, PsiH, PsiK and PsiM, creating the recombinant host organism of claim 22; and

synthesizing psilocybin in the recombinant host organism.