CN113817770A

CN113817770A - Construction method and application of CD73 gene humanized non-human animal

Info

Publication number: CN113817770A
Application number: CN202110933098.6A
Authority: CN
Inventors: 沈月雷; 黄蕤; 白阳; 郭朝设; 张美玲; 尚诚彰
Original assignee: Baccetus Beijing Pharmaceutical Technology Co ltd
Current assignee: Baccetus Beijing Pharmaceutical Technology Co ltd
Priority date: 2020-08-12
Filing date: 2021-08-12
Publication date: 2021-12-21
Anticipated expiration: 2041-08-12
Also published as: CN113817770B

Abstract

The invention relates to a humanized gene modified non-human animal, in particular to a genetically modified rodent, especially a genetically modified mouse, and specifically relates to a construction method of an animal model expressing humanized CD73 protein and application thereof in the field of biological medicines.

Description

Construction method and application of CD73 gene humanized non-human animal

Technical Field

The invention belongs to the field of animal genetic engineering and genetic modification, and particularly relates to a CD73 gene humanized non-human animal, a construction method thereof and application thereof in the field of biomedicine.

Background

Immunotherapy, which attacks and kills cancer cells by activating the immune system, is an important area of tumor research in recent years. Some drugs related to tumor immunotherapy are already used for treating cancers at present, and drugs are available on the market and applied to multiple indications, for example, monoclonal antibodies targeting T cell co-stimulatory molecules CTLA-4, PD-1 and ligands thereof have already achieved definite curative effects, but the average response rate of patients is low. Considering that tumor development is a combined effect of multiple links and multiple factors, a single immunotherapy strategy has limited treatment effect, and a large number of patients can generate drug resistance to treatment, and the clinical combination of two or more immunotherapy approaches is a trend of tumor treatment. The development of more drugs which can be used for improving the tumor recognition and killing ability of the immune system is one of the hot spots of immunological research.

CD73 is an extracellular-5' -nucleotidase encoded by NT5E gene (also known as CD73 gene), has a protein molecular weight of 70KD, is widely expressed on the cell surface of human endothelial cells and lymphocytes such as Tregs (suppressor T cells), and is anchored on the cell surface by Glucosylphosphatidyinositol (GPI) to form a homodimer, but can also be cleaved and dissociated in the circulatory system. CD37 at the upstream of CD73 converts ATP with immune activation into Adenosine Monophosphate (AMP), the generated AMP is converted into adenosine by CD73, adenosine can bind with a downstream adenosine receptor, and then a series of signal paths related to immune activation, such as LCK, MAPK, PKC and the like, are inhibited through adenosine signal, so that the immune inhibition effect is exerted, and the ATP is a self-protection mechanism of a human body.

Tumor Microenvironment (TME) is composed of various cells, matrix, enzymes, cytokines, metabolites, etc., has the characteristics of significant hypoxia, low pH and high pressure, and has a great difference from normal tissues. CD73 is a typical Hypoxia Inducible Factor (HIF) target gene, and HIF-1 is induced by tumor hypoxia environment, so that the expression of CD73 in the tumor microenvironment is remarkably increased, adenosine is further catalyzed, and the immune system is inhibited, and the occurrence and the development of tumors are promoted. The research finds that the expression of CD73 is increased in various tumors, and the prognosis of non-small cell lung cancer patients with high expression of CD73 is poor, which indicates that tumor cells may utilize a CD73 mechanism to generate immune escape. Preclinical studies have shown that targeting CD73 can produce good antitumor effects (Stagg J, et al. anti-CD73 antibody therapy inhibition breakthrough growth and metastasis. proc Natl Acad Sci U S a (2010)107: 1547-52.). In addition, treatment regimens such as PD-1/PD-L1 monoclonal antibody, chemotherapy, etc. may cause elevated expression of CD73 in the tumor microenvironment, which in turn inhibits the immune response (i.e., acquired resistance) through adenosine signaling, and combining CD73 blocking therapy with other immune molecule modulators (e.g., CTLA-4 antibody, PD-1 antibody) helps to achieve better efficacy, an attractive option (Beavis PA, et al. Oncoimmoniology.2015; 4(11): e 1046675.).

Due to the fact that the target mechanism is clear and the preclinical research result is remarkable, at present, more than ten enterprises around the world develop CD73 antibody drugs, and a plurality of projects enter the early clinical stage, for example, the CD 73-inhibited monoclonal antibody BMS-986179 of BMS is in phase I clinical research, and the indication is late solid tumor; the CD 73-inhibiting monoclonal antibody oleclumab developed by AZ/MedImmune has entered phase II clinical studies on recurrent ovarian cancer. In addition, some small molecule compounds aiming at the target, such as a biotechnology innovation company named as Arcus Biosciences which is awarded in the 11-month-repudiation of 1.07 billion dollars in google 2017, one of the major products of the company is a small molecule drug AB680 targeting CD73, and the early clinical research is completed and the clinical examination and approval is waited. Considering that no medicine aiming at the target spot is on the market at present, the existing medicine can not meet the clinical requirement, and more companies are expected to be put into the medicine development of the target spot in the future along with the deep research.

The experimental animal disease model is an indispensable research tool for researching etiology and pathogenesis of human diseases, developing prevention and treatment technologies and developing medicines. The existing animal model related to CD73 is mainly a knockout mouse, and is mainly used for researching CD73, adenosine biological functions (genotype, function and regulation) and related disease mechanisms. For example, Hayo Castrop et al (2004) to investigate whether adenosine formation is dependent on extracellular nucleotide hydrolysis, a CD 73-deficient mouse was prepared by disrupting exons 2 and 3 of the CD73 gene, which was normal in behavior and fertility compared to wild-type mice, but with a significantly reduced ability to constrict glomerular arterioles upon Henle perfusion, concluding that adenosine production on the glomeruli was largely dependent on CD 73-mediated dephosphorylation of AMP; patrycja Koszaka et al (2004) to investigate the effect of extracellular adenosine formation in vascular homeostasis, CD73 knockout mice lacking exon 2 were prepared, demonstrating that adenosine formation outside CD73 cells can modulate coronary vascular tone and inhibit platelet activation. Linda f.thompson et al (2004) to investigate whether CD73 could functionally regulate vascular permeability during basal conditions or hypoxia, mice with exon 3 deletions of the CD73 gene were prepared and confirmed that CD73 is a key regulator of cardiovascular permeability in vivo. (Hayo Castop, et al J Clin invest.2004Sep; 114(5): 634-42.; Koszaka P, et al circulation research.2004; 95: 814-. In conclusion, the existing model is mainly used for CD73, adenosine biological function (genotype, function, regulation) and related disease mechanism research.

CD73 functions by forming homodimers, each monomer comprising a binding site for an N-terminal domain and a C-terminal domain, the two domains being linked by a short helix, and the two monomers being anchored to the plasma membrane after being linked by a non-covalent bond and a GPI linkage via a C-terminal serine residue. In the mature form, a 26 amino acid signal peptide in the N-terminal domain coordinates to catalyze divalent zinc, cobalt, and other metal ions, while the C-terminal domain is primarily the binding site for AMPs. In the development process of CD73 target related drugs, due to species differences between human and mouse, the accuracy of antibody binding with target protein sometimes reaches a single amino acid change in protein (Joseph Conlon J, et al. Immunol.2013May 15; 190(10): 5216-. At present, an in-vivo model widely used for researching the drug effect of an antibody drug is a human tumor xenograft mouse model, but the targeting property and the specificity of the model in the research of a specific target are not strong, so the accuracy of the result of the research on the drug effect of the model is not high.

A gene humanized animal model, namely, a normal or mutant gene animal model which is closer to the physiological or disease characteristics of a human can be established by replacing homologous genes of an animal genome with human normal or mutant genes by using a gene editing technology. The gene humanized animal not only has important application value, for example, the humanized animal model of cell or tissue transplantation can be improved and promoted by gene humanization, but also more importantly, the human protein can be expressed or partially expressed in the animal body due to the insertion of the human gene segment, and the gene humanized animal can be used as a target of a medicine only capable of identifying the amino acid sequence of the human protein, thereby providing possibility for screening anti-human antibodies and other medicines at the animal level. However, due to differences in physiology and pathology between animals and humans, coupled with the complexity of genes (i.e., genetic factors), how to construct an "effective" humanized animal model for new Drug development remains the greatest challenge (Scheer N, Snaith M, Wolf CR, Seibler J. Generation and compliance of genetic humanized models, Drug Discov Today; 18(23-24):1200,11,2013).

Disclosure of Invention

In a first aspect of the invention, a method for constructing a non-human animal humanized with a CD73 gene, wherein the non-human animal expresses a human or humanized CD73 protein.

Preferably, the non-human animal has reduced or absent expression of endogenous CD73 protein.

Preferably, the humanized CD73 protein comprises all or part of a human CD73 protein. Further preferably comprises all or part of the amino acid sequence encoded by exons 1 to 9 of the human CD73 gene.

In one embodiment of the present invention, the amino acid sequence of the humanized CD73 protein comprises one of the following groups:

a) SEQ ID NO:4, or a portion or all of the amino acid sequence set forth in seq id no;

b) and SEQ ID NO:4 is at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99%;

c) and SEQ ID NO:4 differ by no more than 10, 9, 8, 7, 6, 5, 4, 3, 2, or by no more than 1 amino acid; or the like, or, alternatively,

d) and SEQ ID NO:4, including substitution, deletion and/or insertion of one or more amino acid residues.

Preferably, the genome of the non-human animal comprises a human or humanized CD73 gene.

Preferably, the genome of the non-human animal comprises all or part of exons 1 to 9 of the human CD73 gene. Further preferably, the gene comprises one or a combination of two or more exons from exon 1 to exon 9 of human CD 73.

In one embodiment of the present invention, exon 1, exon 2, exon 3, exon 4, exon 5, exon 6, exon 7, exon 8 and exon 9 of the human CD73 gene are included.

Preferably, all or part of exon 1, all of exons 2 to 8 and all or part of exon 9 are contained, preferably also intron 1-2 and/or intron 8-9, wherein part of exon 1 comprises at least a nucleotide sequence of 100bp, for example at least a nucleotide sequence of 100, 150, 200, 250, 300, 310, 330, 335, 336, 337, 338, 339, 340, 350, 370, 390, 396bp, further preferably a nucleotide sequence of 339 or 396 bp; part of exon 1 comprises the nucleotide sequence from the start codon of exon 1 to the last nucleotide of exon 1, and part of exon 9 comprises at least 50bp, for example at least 50, 100, 150, 160, 161, 162, 163, 164, 165, 170, 190, 200, 500, 700, 900, 1000, 1500, 1700, 1900, 1944bp, further preferably 164 or 1944 bp; the portion of exon 9 comprises from the first nucleotide of exon 9 to the stop codon.

In a particular embodiment of the invention, all or part of the CDs sequence of the human CD73 gene or all or part of the full-length nucleotide sequence of human CD73 is comprised.

In one embodiment of the invention, the non-human animal comprises SEQ ID NO:9 or 17.

Preferably, the non-human animal comprises all or part of a nucleotide sequence encoding human CD73 protein. Further preferred comprises a nucleic acid sequence encoding SEQ ID NO: 4.

In one embodiment of the present invention, the humanized CD73 gene comprises one of the following groups:

A) SEQ ID NO:9 or SEQ ID NO:17, all or part of a nucleotide sequence set forth in seq id no;

B) and SEQ ID NO:9 or SEQ ID NO:17 is at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% identical;

C) and SEQ ID NO:9 or SEQ ID NO:17 differ by no more than 10, 9, 8, 7, 6, 5, 4, 3, 2, or no more than 1 nucleotide; or the like, or, alternatively,

D) and SEQ ID NO:9 or SEQ ID NO:17, comprising a nucleotide sequence in which one or more nucleotide residues are substituted, deleted and/or inserted.

Preferably, the humanized CD73 gene further comprises a nucleotide sequence encoding a 2A peptide, preferably a nucleotide sequence encoding P2A or T2A. Further preferred comprises SEQ ID NO:10, or a nucleotide sequence shown in the figure.

Preferably, the humanized CD73 gene further comprises a nucleotide sequence encoding SEQ ID NO: 11.

Preferably, the humanized CD73 gene further comprises a transcription termination sequence. Further, one or a combination of two or more of 3' UTR, polyA, WPRE, STOP, and lox2 is preferable. Still further preferred are compounds comprising a STOP sequence as set forth in SEQ ID NO: 20 and/or the 3' UTR sequence is as shown in SEQ ID NO: 22.

Preferably, the humanized CD73 gene further comprises a Neo cassette.

Preferably, the humanized CD73 gene further comprises endogenous regulatory sequences. More preferably, an endogenous promoter nucleotide sequence is included.

In one embodiment of the present invention, the humanized CD73 gene comprises one of the following groups in 5 'to 3' order:

A) encoding the amino acid sequence of SEQ ID NO: 11, a nucleotide sequence encoding a 2A peptide, a CDs sequence of the human CD73 gene, and a transcription termination sequence;

B) encoding the amino acid sequence of SEQ ID NO: 11, a nucleotide sequence encoding a 2A peptide, the CDs sequence of the human CD73 gene, a transcription termination sequence, and a Neo cassette;

C) a portion of the CDs sequence of the human CD73 gene and/or the full-length nucleotide sequence of human CD73 and a transcription termination sequence;

D) part of the CDs sequence of the human CD73 gene and/or the full-length nucleotide sequence of human CD73, a transcription termination sequence and the Neo cassette;

E) an endogenous regulatory sequence encoding SEQ ID NO: 11, a nucleotide sequence encoding a 2A peptide, a CDs sequence of the human CD73 gene, and a transcription termination sequence;

F) an endogenous regulatory sequence encoding SEQ ID NO: 11, a nucleotide sequence encoding a 2A peptide, the CDs sequence of the human CD73 gene, a transcription termination sequence, and a Neo cassette;

G) endogenous regulatory sequences, the CDs sequence of the human CD73 gene and/or portions of the full-length nucleotide sequence of human CD73 and transcription termination sequences;

H) endogenous regulatory sequences, the CDs sequence of the human CD73 gene and/or part of the full-length nucleotide sequence of human CD73, transcription termination sequences and Neo-cassette; or the like, or, alternatively,

I) a nucleotide sequence encoding the 2A peptide and a portion of the full-length nucleotide sequence of human CD 73.

Preferably, the humanized CD73 gene comprises SEQ ID NO: 23.

(i) comprises the amino acid sequence of SEQ ID NO: 12. 13 or 18, or a portion or all of the nucleotide sequence set forth in seq id no;

(ii) comprises a nucleotide sequence substantially identical to SEQ ID NO: 12. 13 or 18 is at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or at least 99%;

(iii) comprises a nucleotide sequence substantially identical to SEQ ID NO: 12. 13 or 18 differ by no more than 10, 9, 8, 7, 6, 5, 4, 3, 2, or no more than 1 nucleotide; or the like, or, alternatively,

(iv) comprises a nucleotide sequence substantially identical to SEQ ID NO: 12. 13 or 18, including nucleotide sequences with one or more nucleotides substituted, deleted and/or inserted.

In one embodiment of the present invention, the mRNA transcribed from the humanized CD73 gene comprises one of the following groups:

(i) comprises the amino acid sequence of SEQ ID NO:19, or a portion or all of the nucleotide sequence set forth in seq id no;

(ii) comprises a nucleotide sequence substantially identical to SEQ ID NO:19 is at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or at least 99% identical;

(iii) comprises a nucleotide sequence substantially identical to SEQ ID NO:19 by no more than 10, 9, 8, 7, 6, 5, 4, 3, 2, or by no more than 1 nucleotide; or the like, or, alternatively,

(iv) comprises a nucleotide sequence substantially identical to SEQ ID NO:19, including nucleotide sequences with one or more nucleotides substituted, deleted and/or inserted.

Preferably, the genome of the non-human animal further comprises other gene modifications, and further preferably, the other genes comprise at least one of PD-1, PD-L1, CTLA-4, LAG-3, BTLA, CD27, CD28, CD47, CD137, CD154, OX40, sirpa, TIGIT, TIM-3, CD40, and GITR genes.

Preferably, the construction method comprises introducing a nucleotide sequence comprising a human or humanized CD73 gene into the non-human animal CD73 locus. It is further preferred that all or part of exons 1 to 9 comprising the human CD73 gene be introduced into the non-human animal CD73 locus.

In one embodiment of the invention, exon 1, exon 2, exon 3, exon 4, exon 5, exon 6, exon 7, exon 8, and exon 9 of the gene comprising human CD73 are introduced into the non-human animal CD73 locus.

Preferably, all or part of exon 1, all of exons 2 to 8 and all or part of exon 9 are included, preferably further including intron 1-2 and/or intron 8-9, into the CD73 locus of the non-human animal, wherein part of exon 1 comprises at least 100bp of nucleotide sequence, for example at least 100, 150, 200, 250, 300, 310, 330, 335, 336, 337, 338, 339, 340, 350, 370, 390, 396bp of nucleotide sequence, further preferably 339 or 396bp of nucleotide sequence; part of exon 1 comprises the nucleotide sequence from the start codon of exon 1 to the last nucleotide of exon 1, and part of exon 9 comprises at least 50bp, for example at least 50, 100, 150, 160, 161, 162, 163, 164, 165, 170, 190, 200, 500, 700, 900, 1000, 1500, 1700, 1900, 1944bp, further preferably 164 or 1944 bp; the portion of exon 9 comprises from the first nucleotide of exon 9 to the stop codon.

In one embodiment of the invention, the method of construction comprises introducing into the non-human animal CD73 locus all or part of the CDs sequence comprising human CD73 gene or all or part of the full-length nucleotide sequence of human CD 73.

Preferably, the construction method comprises introducing a cDNA sequence comprising the human CD73 gene into the non-human animal CD73 locus.

In one embodiment of the invention, the method of construction comprises the step of using a nucleic acid comprising SEQ ID NO: 9. 12, 13, 17 or 18 into the non-human animal CD73 locus.

Preferably, the construction method comprises introducing the nucleotide sequence comprising all or part of the nucleotide sequence encoding human CD73 protein into the CD73 locus of the non-human animal. Further preferred is a polypeptide comprising a sequence encoding SEQ ID NO:4 into the non-human animal CD73 locus.

Preferably, the nucleotide sequence of the human or humanized CD73 gene is operably linked to a non-human animal endogenous regulatory element. Further preferably to a non-human animal promoter.

Preferably, the location of said introduction is after the endogenous regulatory elements of the CD73 gene. Preferably, the endogenous regulatory element is an endogenous promoter.

Preferably, the site of introduction is located on exons 1 to 9 of the non-human animal CD73 gene. Further preferably located from the start codon to the stop codon of the CD73 gene or the full-length nucleotide sequence of the non-human animal CD73 gene.

Preferably, the site of introduction is located on exons 2 to 9 of the non-human animal CD73 gene. Further preferably on the exon No. 2 and/or the intron No. 2-3.

Preferably, the introduction described herein includes, but is not limited to, insertion, substitution, or transgene.

Preferably, the non-human animal is homozygous or heterozygous.

Preferably, the genome of the non-human animal comprises a humanized CD73 gene on at least one chromosome.

Preferably, at least one cell in the non-human animal expresses a human or humanized CD73 protein.

Preferably, the non-human animal is constructed using gene editing techniques including gene targeting using embryonic stem cells, regular clustered spacer short palindromic repeats (CRISPR/Cas9) techniques, Zinc Finger Nucleases (ZFNs) techniques, transcription activator-like effector nucleases (TALENs) techniques, homing endonucleases (megabase megaribozymes), or other molecular biology techniques.

Preferably, the construction of the non-human animal is performed using a targeting vector, wherein the targeting vector comprises a portion of the human CD73 gene. Further preferably, the gene comprises all or part of exons 1 to 9 of human CD 73. Still further preferred is a combination comprising one or more than two exons among exons 1 to 9 of human CD73 gene.

Preferably comprises all or part of exon 1, all of exons 2 to 8 and all or part of exon 9, more preferably also comprises intron 1-2 and/or intron 8-9, wherein part of exon 1 comprises at least a nucleotide sequence of 100bp, for example at least a nucleotide sequence of 100, 150, 200, 250, 300, 310, 330, 335, 336, 337, 338, 339, 340, 350, 370, 390, 396bp, further preferably a nucleotide sequence of 339 or 396 bp; part of exon 1 comprises the nucleotide sequence from the start codon of exon 1 to the last nucleotide of exon 1, and part of exon 9 comprises at least 50bp, for example at least 50, 100, 150, 160, 161, 162, 163, 164, 165, 170, 190, 200, 500, 700, 900, 1000, 1500, 1700, 1900, 1944bp, further preferably 164 or 1944 bp; the portion of exon 9 comprises from the first nucleotide of exon 9 to the stop codon.

In one embodiment of the invention, the polypeptide comprises a nucleotide sequence encoding SEQ ID NO: 4.

In one embodiment of the invention, the polypeptide comprising SEQ ID NO: 9. 12, 13, 17 or 18.

Preferably, the targeting vector further comprises a5 'arm (5' homology arm) and/or a3 'arm (3' homology arm). The 5' arm is selected from 100-10000 nucleotides in length of the non-human animal CD73 gene genome DNA. Preferably, the 5' arm has at least 90% homology with NCBI accession number NC _ 000075.6. Further preferably, the 5' arm sequence is identical to SEQ ID NO: 5. 7 or 15 have at least 90% homology. The 3' arm is selected from 100-10000 nucleotides in length of the non-human animal CD73 gene genome DNA. Preferably, the 3' arm has at least 90% homology with NCBI accession number NC _ 000075.6. Further preferably, the 3' arm sequence is identical to SEQ ID NO: 6. 8 or 16 have at least 90% homology.

Preferably, the 5 'arm is a DNA fragment homologous to the 5' end of the transition region to be altered.

Preferably, the 3 'arm is a DNA fragment homologous to the 3' end of the transition region to be altered.

Preferably, the transition region to be altered is located in exon 1 to exon 9 of the non-human animal CD73 gene.

In one embodiment of the invention, the gene is located in exon 2 and/or intron 2-3 of the non-human animal CD73 gene.

In one embodiment of the invention, the nucleotide sequence is located from the start codon to the stop codon of the non-human animal CD73 gene.

In a specific embodiment of the invention, the construction method comprises introducing the targeting vector into a cell of a non-human animal, culturing the cell (preferably an embryonic stem cell), transplanting the cultured cell into an oviduct of a female non-human animal, allowing the female non-human animal to develop, and identifying and screening the non-human animal humanized with the CD73 gene.

Preferably, to improve recombination efficiency, a non-human animal can also be constructed using sgRNA targeting the CD73 gene together with the above-described targeting vector. Wherein the sgRNA targets the non-human animal CD73 gene, while the sequence of the sgRNA is on the target sequence on the CD73 gene to be altered.

Preferably, the target site of the sgRNA is located on exon 1 to exon 9 sequences of the CD73 gene. Further preferably, the target site of the sgRNA is located on exon 2, intron 2-3, and/or exon 3 sequences of the CD73 gene.

In a specific embodiment of the invention, the target sites of the sgrnas are located on exon 2 and intron 2-3 of the CD73 gene.

Preferably, the sequence of the target site of the sgRNA is as shown in SEQ ID NO: 26-41.

In a specific embodiment of the invention, the construction method comprises the steps of introducing the targeting vector, the sgRNA targeting the CD73 gene and the Cas9 into a non-human animal cell, culturing the cell (preferably an embryonic stem cell), transplanting the cultured cell into an oviduct of a female non-human animal, allowing the female non-human animal to develop, and identifying and screening the non-human animal humanized with the CD73 gene.

Preferably, the non-human animal can be selected from any non-human animal such as rodent, pig, chicken, rabbit, monkey, etc. which can be genetically modified to produce a humanized gene.

Preferably, the non-human animal is a non-human mammal. Further preferably, the non-human mammal is a rodent. Still more preferably, the rodent is a rat or a mouse.

Preferably, the non-human animal is an immunodeficient non-human mammal. Further preferably, the immunodeficient non-human mammal is an immunodeficient rodent, an immunodeficient pig, an immunodeficient rabbit or an immunodeficient monkey. Still further preferably, the immunodeficient rodent is an immunodeficient mouse or rat. Most preferably, the immunodeficient mouse is NOD-Prkdc^scid IL-2rγ^nullMouse, NOD-Rag 1^-/--IL2rg^-/-(NRG) mice, Rag 2^-/--IL2rg^-/-(RG) mice, NOD/SCID mice or nude mice.

In a second aspect of the invention, there is provided a non-human animal humanized with a CD73 gene, the genome of which comprises a human or humanized CD73 gene.

Preferably, all or part of exon 1, all of exon 2 to 8 and all or part of exon 9 are contained, preferably also the intron 1-2 and/or the intron 8-9, wherein the part of exon 1 comprises the first nucleotide of exon 1 to the last nucleotide of exon 1 and the part of exon 9 comprises the first nucleotide of exon 9 to the stop codon.

Preferably, the humanized CD73 gene further comprises a 2A peptide, preferably P2A or T2A. Further preferred comprises SEQ ID NO:10, or a nucleotide sequence shown in the figure.

Preferably, the humanized CD73 gene further comprises a transcription termination sequence. Further preferred is one or a combination of two or more of 3' UTR, polyA, WPRE, and lox 2. Even more preferably comprises SEQ ID NO: 20 and/or SEQ ID NO: 22.

Preferably, the humanized CD73 gene further comprises a Neo cassette.

Preferably, the humanized CD73 gene comprises SEQ ID NO: 23.

Preferably, the non-human animal body expresses a human or humanized CD73 protein.

Preferably, the non-human animal can be selected from any non-human animal such as rodent, pig, rabbit, monkey, etc. which can be genetically modified by gene editing.

In a third aspect of the invention, there is provided a targeting vector comprising a portion of the human CD73 gene.

Preferably, the targeting vector comprises all or part of exons 1 to 9 of the human CD73 gene. Further preferred is a combination comprising one or more than two exons among exons 1 to 9 of human CD73 gene.

Preferably, the targeting vector further comprises a marker gene. Further preferably, the marker gene is a gene encoding a negative selection marker. Still more preferably, the gene encoding the negative selection marker is a gene encoding diphtheria toxin subunit a (DTA).

In one embodiment of the present invention, the targeting vector further comprises a resistance gene for positive clone selection. Further preferably, the resistance gene selected by the positive clone is neomycin phosphotransferase coding sequence Neo.

In one embodiment of the present invention, the targeting vector further comprises a specific recombination system. Further preferably, the specific recombination system is a Frt recombination site (a conventional LoxP recombination system can also be selected). The specific recombination system is provided with two Frt recombination sites which are respectively connected to two sides of the resistance gene.

In a fourth aspect of the invention, there is provided a sgRNA that targets a non-human animal CD73 gene, while the sequence of the sgRNA is on a target sequence on a CD73 gene to be altered.

In a fifth aspect of the invention, a DNA molecule encoding the sgRNA described above is provided. Preferably, the double strands of the DNA molecules are the upstream and downstream sequences of the sgRNA, or the forward and reverse oligonucleotide sequences after the addition of the enzyme cleavage site.

In one embodiment of the present invention, the double strands of the DNA molecule are SEQ ID NOs: 32 and SEQ ID NO: 42.

in one embodiment of the present invention, the double strands of the DNA molecule are SEQ ID NOs: 39 and SEQ ID NO: 45.

in one embodiment of the present invention, the double strands of the DNA molecule are SEQ ID NOs: 43 and SEQ ID NO: 44.

in one embodiment of the present invention, the double strands of the DNA molecule are SEQ ID NOs: 46 and SEQ ID NO: 47.

in a sixth aspect of the present invention, a sgRNA vector is provided, which includes the sgRNA described above.

In a seventh aspect of the present invention, there is provided a cell comprising the targeting vector, the sgRNA, the DNA molecule, or the sgRNA vector.

In an eighth aspect of the present invention, there is provided a use of the targeting vector, the sgRNA, the DNA molecule, the sgRNA vector, or the cell for CD73 gene modification. Preferably comprising use in knock-out, insertion or substitution of the CD73 gene.

In the ninth aspect of the invention, a humanized CD73 gene is provided, wherein the humanized CD73 gene comprises a part of a human CD73 gene.

Preferably, the humanized CD73 gene comprises all or part of exons 1 to 9 of human CD73 gene. Further preferably, the gene comprises one or a combination of two or more exons from exon 1 to exon 9 of human CD 73.

Preferably, all or part of exon 1, all of exons 2 to 8 and all or part of exon 9 are contained, more preferably, the intron 1-2 and/or the intron 8-9 are also contained, wherein the part of exon 1 at least comprises a nucleotide sequence of 100bp, for example, at least comprises a nucleotide sequence of 100, 150, 200, 250, 300, 310, 330, 335, 336, 337, 338, 339, 340, 350, 370, 390, 396bp, further preferably, comprises a nucleotide sequence of 339 or 396 bp; part of exon 1 comprises the nucleotide sequence from the start codon of exon 1 to the last nucleotide of exon 1, and part of exon 9 comprises at least 50bp, for example at least 50, 100, 150, 160, 161, 162, 163, 164, 165, 170, 190, 200, 500, 700, 900, 1000, 1500, 1700, 1900, 1944bp, further preferably 164 or 1944 bp; the portion of exon 9 comprises from the first nucleotide of exon 9 to the stop codon.

Preferably, the humanized CD73 gene comprises all or part of a nucleotide sequence encoding human CD73 protein. Further preferred comprises a nucleic acid sequence encoding SEQ ID NO: 4.

Preferably, the humanized CD73 gene further comprises a transcription termination sequence, preferably one or a combination of two or more of 3' UTR, polyA, WPRE, STOP or lox 2. Further preferred are compounds comprising a STOP sequence as set forth in SEQ ID NO: 20 and/or 3' UTR sequences are shown in SEQ ID NO: 22.

Preferably, the humanized CD73 gene further comprises a Neo cassette.

Preferably, the humanized CD73 gene further comprises endogenous regulatory sequences. Further preferably, an endogenous promoter nucleotide sequence is included.

E) an endogenous regulatory sequence encoding SEQ ID NO: 11, a 2A peptide, a CDs sequence of the human CD73 gene, and a transcription termination sequence;

F) an endogenous regulatory sequence encoding SEQ ID NO: 11, 2A peptide, the CDs sequence of the human CD73 gene, a transcription termination sequence, and a Neo cassette;

Preferably, the humanized CD73 gene comprises SEQ ID NO: 23.

In the tenth aspect of the invention, a humanized CD73 protein is provided, wherein the humanized CD73 protein comprises a part of a human CD73 protein.

Preferably, the humanized CD73 protein comprises all or part of an amino acid sequence encoded by exons 1 to 9 of the human CD73 gene. Further preferably, the humanized CD73 protein is encoded by the humanized CD73 gene described above.

In the eleventh aspect of the present invention, there is provided a polygene-modified non-human animal, wherein the non-human animal is the above non-human animal or the non-human animal obtained by the above construction method, and the genome of the non-human animal comprises a modification of at least one of genes PD-1, PD-L1, CTLA-4, LAG-3, BTLA, CD27, CD28, CD47, CD137, CD154, OX40, sirpa, TIGIT, TIM-3, CD40, and GITR.

In a twelfth aspect of the present invention, there is provided a method for constructing a polygene-modified non-human animal, comprising the steps of:

providing the non-human animal or the non-human animal obtained by the construction method;

and (II) mating the non-human animal provided in the step (I) with other genetically modified non-human animals, performing in vitro fertilization or directly performing gene editing, and screening to obtain the multi-gene modified non-human animal.

Preferably, the other genetically modified non-human animal comprises a humanized non-human animal of at least one of the genes PD-1, PD-L1, CTLA-4, LAG-3, BTLA, CD27, CD28, CD47, CD137, CD154, OX40, SIRPa, TIGIT, TIM-3, CD40, and GITR.

Preferably, each of the plurality of genes humanized in the genome of the polygenic modified non-human animal may be homozygous or heterozygous.

In a thirteenth aspect of the present invention, there is provided a non-human animal or a progeny thereof obtained by the above construction method.

In a fourteenth aspect of the present invention, an animal tumor-bearing or inflammation model is provided, wherein the tumor-bearing or inflammation model is derived from the above non-human animal or the non-human animal obtained by the above construction method.

In a fifteenth aspect of the present invention, there is provided a method for constructing a tumor-bearing or inflammation model of an animal, the method comprising the step of obtaining a humanized non-human animal or a polygene-modified non-human animal of the CD73 gene by using the above-described construction method.

In a sixteenth aspect of the present invention, an application of the above non-human animal or the non-human animal obtained by the above construction method in preparing an animal model with tumor or inflammation is provided.

In a seventeenth aspect of the present invention, there is provided a cell or cell line or primary cell culture derived from the above non-human animal or the non-human animal obtained by the above construction method, or the above tumor-bearing or inflammation model. Preferably, the cell or cell line or primary cell culture is not capable of developing into an individual animal.

In an eighteenth aspect of the present invention, there is provided a tissue or organ or culture thereof derived from the above-mentioned non-human animal or the non-human animal obtained by the above-mentioned construction method, or the above-mentioned tumor-bearing or inflammation model. Preferably, the tissue or organ or culture thereof is incapable of developing into an individual animal.

In a nineteenth aspect of the present invention, there is provided a tumor-bearing tissue obtained from the above-mentioned non-human animal or the non-human animal obtained by the above-mentioned construction method, or the above-mentioned tumor-bearing or inflammation model. Preferably, said tumor-bearing tumor tissue is incapable of developing into an individual animal.

In a twentieth aspect of the invention, there is provided a cell humanised with a CD73 gene, said cell comprising a human or humanised CD73 gene. Preferably comprising the humanized CD73 gene described above.

Preferably, the cell expresses a human or humanized CD73 protein. Further preferred is the above-described humanized CD73 protein of the present invention. Preferably, the cells are not capable of developing into an individual animal.

In a twenty-first aspect of the present invention, there is provided a cell deficient in the CD73 gene, said cell being deficient in all or part of exons 1 to 9 of the CD73 gene. Preferably, all or part of exon 2 is deleted.

In a specific embodiment of the invention, the deletion encodes SEQ ID NO:2, nucleotide sequence 180-189.

In another embodiment of the invention, the nucleotide sequence of the CD73 gene from the start codon to the stop codon is deleted.

Preferably, the cells are not capable of developing into an individual animal.

In a twenty-second aspect of the present invention, there is provided a non-human animal deficient in the CD73 gene, said non-human animal being deficient in all or part of exons 1 to 9 of the CD73 gene. Preferably, all or part of exon 2 is deleted.

In a twenty-third aspect of the present invention, there is provided a method for constructing a non-human animal or cell in which the CD73 gene is deleted, the method using the sgRNA described above.

In a twenty-fourth aspect of the invention, there is provided a construct comprising the humanized CD73 gene described above.

Preferably, the construct expresses a human or humanized CD73 protein, and the humanized CD73 protein is the humanized CD73 protein described above.

In a twenty-fifth aspect of the invention, there is provided a cell comprising the above construct.

Preferably, the cells are not capable of developing into an individual animal.

In a twenty-sixth aspect of the invention, there is provided a tissue comprising the above-described cells.

Preferably, the tissue is incapable of developing into an individual animal.

A twenty-seventh aspect of the present invention provides a use of a non-human animal derived from the above-mentioned non-human animal, a non-human animal obtained by the above-mentioned construction method, the above-mentioned humanized CD73 gene, the above-mentioned humanized CD73 protein, the above-mentioned tumor-bearing or inflammatory model, the above-mentioned cell or cell line or primary cell culture, the above-mentioned tissue or organ or culture thereof, the above-mentioned tumor-bearing tissue, the above-mentioned cell, the above-mentioned construct, the above-mentioned cell or the above-mentioned tissue for product development of an immune process involving human cells, for production of an antibody, or as a model system for pharmacological, immunological, microbiological, or medical research; or in the production and use of animal experimental disease models for the development of new diagnostic and/or therapeutic strategies; or screening, verifying, evaluating or researching CD73 channel function, human CD73 channel signal mechanism, human-targeting antibody, human-targeting medicine, medicine effect, immunity-related disease medicine and anti-tumor medicine, screening and evaluating human medicine and medicine effect research.

Preferably, the use is not a method of treatment and/or diagnosis of a disease.

In a twenty-eighth aspect of the invention, there is provided a method of screening for a modulator specific for human CD73, the method comprising administering the modulator to an individual; wherein the individual is selected from the group consisting of the above non-human animal, the non-human animal obtained by the above construction method, and the above tumor-bearing or inflammation model.

Preferably, the screening method further comprises implanting tumor cells into the individual.

Preferably, the modulator is selected from CAR-T, a drug; preferably, the drug is an antibody.

Preferably, the screening method further comprises detecting tumor suppression.

Preferably, the modulator is a monoclonal antibody or a bispecific antibody or a combination of two or more drugs.

Preferably, the detection comprises determining the size and/or proliferation rate of the tumor cells.

Preferably, the detection method comprises vernier caliper measurement, flow cytometry detection and/or animal in vivo imaging detection.

Preferably, the detecting comprises assessing the weight, fat mass, activation pathways, neuroprotective activity or metabolic changes in the individual, including changes in food consumption or water consumption.

Preferably, the tumor cell is derived from a human or non-human animal.

Preferably, the screening method is not a therapeutic method. The method is used for screening or evaluating drugs, and detecting and comparing the drug effects of candidate drugs to determine which candidate drugs can be used as drugs and which can not be used as drugs, or comparing the drug effect sensitivity degrees of different drugs, namely, the treatment effect is not necessary and is only a possibility.

In a twenty-ninth aspect of the present invention, there is provided an evaluation method of an intervention program, the evaluation method comprising implanting tumor cells into an individual, applying the intervention program to the individual in which the tumor cells are implanted, and detecting and evaluating a tumor suppression effect of the individual to which the intervention program is applied; wherein the individual is selected from the group consisting of the above-mentioned non-human animal, the non-human animal obtained by the above-mentioned construction method, the above-mentioned non-human animal or a progeny thereof, or the above-mentioned tumor-bearing or inflammation model.

Preferably, the intervention regimen is selected from CAR-T, drug therapy. Further preferably, the drug is an antigen binding protein. The antibody binding protein is an antibody.

Preferably, the tumor cell is derived from a human or non-human animal.

Preferably, the method of assessing the intervention regimen is not a method of treatment. The evaluation method detects and evaluates the effect of the intervention program to determine whether the intervention program has a therapeutic effect, i.e. the therapeutic effect is not necessarily but only a possibility.

In a thirtieth aspect of the present invention, there is provided a use of the non-human animal derived from the above non-human animal, the non-human animal obtained by the above construction method, the above non-human animal or its progeny, the above tumor-bearing or inflammation model for preparing a human CD 73-specific modulator.

In a thirty-first aspect of the present invention, there is provided a use of the non-human animal obtained by the above-mentioned construction method, the above-mentioned non-human animal or its progeny, and the above-mentioned tumor-bearing or inflammation model in the preparation of a medicament for treating tumor or autoimmune disease.

The CD73 gene humanized non-human animal can normally express human or humanized CD73 protein in vivo, can be used for drug screening, drug effect evaluation, autoimmune diseases and tumor treatment aiming at human CD73 pathway target sites, can accelerate the development process of new drugs, and can save time and cost.

The "immune-related diseases" described in the present invention include, but are not limited to, allergy, asthma, dermatitis, myocarditis, nephritis, hepatitis, systemic lupus erythematosus, rheumatoid arthritis, scleroderma, hyperthyroidism, idiopathic thrombocytopenic purpura, autoimmune hemolytic anemia, ulcerative colitis, autoimmune liver disease, diabetes, pain, or neurological disorder, etc.

The "tumor" according to the present invention includes, but is not limited to, lymphoma, brain cancer, non-small cell lung cancer, cervical cancer, esophageal cancer, leukemia, ovarian cancer, nasopharyngeal cancer, breast cancer, endometrial cancer, colon cancer, rectal cancer, stomach cancer, bladder cancer, lung cancer, bronchial cancer, bone cancer, prostate cancer, pancreatic cancer, liver and bile duct cancer, esophageal cancer, kidney cancer, thyroid cancer, head and neck cancer, testicular cancer, glioblastoma, astrocytoma, melanoma, myelodysplastic syndrome, and sarcoma. Wherein the leukemia is selected from acute lymphocytic (lymphoblastic) leukemia, acute myelogenous leukemia, chronic lymphocytic leukemia, multiple myeloma, plasma cell leukemia, and chronic myelogenous leukemia; said lymphoma is selected from Hodgkin's lymphoma and non-Hodgkin's lymphoma, including B-cell lymphoma, diffuse large B-cell lymphoma, follicular lymphoma, mantle cell lymphoma, marginal zone B-cell lymphoma, T-cell lymphoma, and Waldenstrom's macroglobulinemia; the sarcoma is selected from osteosarcoma, Ewing's sarcoma, leiomyosarcoma, synovial sarcoma, soft tissue sarcoma, angiosarcoma, liposarcoma, fibrosarcoma, rhabdomyosarcoma, and chondrosarcoma.

The invention relates to a whole or part, wherein the whole is a whole, and the part is a part of the whole or an individual forming the whole.

The humanized CD73 protein comprises a part derived from human CD73 protein and a part derived from non-human CD73 protein. Wherein, the humanized CD73 protein comprises 5 to 574 continuous or alternate amino acid sequences which are consistent with the amino acid sequence of the human CD73 protein.

The "humanized CD73 gene" of the present invention comprises a part derived from human CD73 gene and a part derived from non-human CD73 gene. Wherein, the 'humanized CD73 gene' comprises a continuous or alternate 20-46000bp nucleotide sequence which is consistent with the nucleotide sequence of the human CD73 gene, preferably a continuous or alternate 20-1725, 20-45928bp nucleotide sequence, more preferably 20, 50, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1725, 1800, 1900, 2000, 2500, 3000, 5000, 10000, 20000, 30000, 40000, 45000 or 45928bp nucleotide sequence which is consistent with the nucleotide sequence of the human CD73 gene.

The "exon 1 to 9" or "all of the exons 1 to 9" described in the present invention includes nucleotide sequences of exons and introns therebetween, i.e., exon 1, intron 1-2, exon 2, intron 2-3, exon 3, intron 3-4, exon 4, intron 4-5, exon 5, intron 5-6, exon 6, intron 6-7, exon 7, intron 7-8, exon 8, intron 8-9, and exon 9.

The "intron 1-2" described in the present invention means an intron between the exon 1 and the exon 2.

The "locus" of the present invention refers to the position of a gene on a chromosome in a broad sense and refers to a DNA fragment of a certain gene in a narrow sense, and the gene may be a single gene or a part of a single gene. For example, the "CD 73 locus" refers to a DNA fragment of any one of exons 1 to 9 of CD73 gene. In one embodiment of the invention, the introduced CD73 locus may be a DNA fragment of an optional stretch of exons 1 to 9 of the CD73 gene.

The "nucleotide sequence" of the present invention includes a natural or modified ribonucleotide sequence and a deoxyribonucleotide sequence. Preferably DNA, cDNA, pre-mRNA, rRNA, hnRNA, miRNAs, scRNA, snRNA, siRNA, sgRNA, tRNA.

The term "treating" (or "treatment") as used herein means slowing, interrupting, arresting, controlling, stopping, alleviating, or reversing the progression or severity of one sign, symptom, disorder, condition, or disease, but does not necessarily refer to the complete elimination of all disease-related signs, symptoms, conditions, or disorders. The term "treatment" or the like refers to a therapeutic intervention that ameliorates the signs, symptoms, etc. of a disease or pathological state after the disease has begun to develop.

The term "homology" as used herein refers to the fact that, in the aspect of using an amino acid sequence or a nucleotide sequence, a person skilled in the art can adjust the sequence according to the actual working requirement, so that the used sequence has (including but not limited to) 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 70%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9% identity.

One skilled in the art can determine and compare sequence elements or degrees of identity to distinguish between additional mouse and human sequences.

In one aspect, the non-human animal is a mammal. In one aspect, the non-human animal is a small mammal, such as a rhabdoid. In one embodiment, the non-human animal to which the gene is humanized is a rodent. In one embodiment, the rodent is selected from a mouse, a rat, and a hamster. In one embodiment, the rodent is selected from the murine family. In one embodiment, the genetically modified animal is from the family of cricotidae (e.g., mouse-like hamsters), cricotidae (e.g., hamsters, new world rats and mice, voles), muridae (true mice and rats, gerbils, spiny mice, crow rats), marmoraceae (mountaineers, rock mice, tailed rats, madagaska rats and mice), spiny muridae (e.g., spiny mice), and spale (e.g., mole rats, bamboo rats, and zokors). In a particular embodiment, the genetically modified rodent is selected from a true mouse or rat (superfamily murinus), a gerbil, a spiny mouse, and a crowned rat. In one embodiment, the genetically modified mouse is from a member of the murine family. In one embodiment, the animal is a rodent. In a particular embodiment, the rodent is selected from a mouse and a rat. In one embodiment, the non-human animal is a mouse.

In a particular embodiment, the non-human animal is a rodent, a strain of C57BL, C58, a/Br, CBA/Ca, CBA/J, CBA/CBA/mouse selected from BALB/C, a/He, a/J, A/WySN, AKR/A, AKR/J, AKR/N, TA1, TA2, RF, SWR, C3H, C57BR, SJL, C57L, DBA/2, KM, NIH, ICR, CFW, FACA, C57BL/A, C57BL/An, C57BL/GrFa, C57BL/KaLwN, C57BL/6, C57BL/6J, C57BL/6ByJ, C57BL/6NJ, C57BL/10, C57BL/10 sn, C57BL/10Cr and C57 BL/Ola.

The practice of the present invention will employ, unless otherwise indicated, conventional techniques of cell biology, cell culture, molecular biology, transgenic biology, microbiology, recombinant DNA, and immunology. These techniques are explained in detail in the following documents. For example: molecular Cloning A Laboratory Manual, 2nd Ed., ed.by Sambrook, FritschandManiatis (Cold Spring Harbor Laboratory Press: 1989); DNA Cloning, Volumes I and II (d.n. glovered., 1985); oligonucleotide Synthesis (m.j. gaited., 1984); mulliserial.u.s.pat.no. 4, 683, 195; nucleic Acid Hybridization (B.D. Hames & S.J. Higgins.1984); transformation And transformation (B.D. Hames & S.J. Higgins.1984); culture Of Animal Cells (r.i. freshney, alanr.liss, inc., 1987); immobilized Cells And Enzymes (IRL Press, 1986); B.Perbal, A Practical Guide To Molecular Cloning (1984); the series, Methods In ENZYMOLOGY (J.Abelson and M.Simon, eds. inchief, Academic Press, Inc., New York), specific, Vols.154and 155(Wuetal. eds.) and Vol.185, "Gene Expression Technology" (D.Goeddel, ed.); gene Transfer Vectors For Mammarian Cells (J.H.Miller and M.P.Caloseds, 1987, Cold Spring Harbor Laboratory); immunochemical Methods In Cell And Molecular Biology (Mayer And Walker, eds., Academic Press, London, 1987); handbook Of Experimental Immunology, Volumes V (d.m.weir and c.c.blackwell, eds., 1986); and Manipulating the Mouse Embryo, (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1986).

All patents and publications mentioned in this specification are herein incorporated in their entirety by reference into the specification, to the same extent as if each individual publication was specifically and individually indicated to be incorporated herein by reference. Those skilled in the art will recognize that certain changes may be made to the invention without departing from the spirit or scope of the invention. The following examples further illustrate the invention in detail and are not to be construed as limiting the scope of the invention or the particular methods described herein.

Drawings

Embodiments of the invention are described in detail below with reference to the attached drawing figures, wherein:

FIG. 1: mouse and human CD73 gene comparison scheme (not to scale).

FIG. 2: CD73 gene humanised mice CD73 gene representation one (not to scale).

FIG. 3: targeting strategies used to prepare humanized mice with the CD73 gene are shown schematically (not to scale).

FIG. 4: southern Blot results, where ZL001 to ZL018 are mouse numbers, M is Marker, WT is wild type control, H₂O is water control.

FIG. 5: f1 mouse tail PCR for identifying somatic cell genotype, wherein 1F 1001-1F 1011 are tail number, WT is wild control, M is Marker, H₂O is water control.

FIG. 6: schematic representation (not to scale) of the FRT recombination process for mice humanized with the CD73 gene;

FIG. 7: targeting strategy used to generate humanized mice with the CD73 gene is shown schematically (not to scale).

FIG. 8: and (5) detecting the activity of the sgRNA, wherein Con is a negative control, and PC is a positive control.

FIG. 9: the genotype identification result of mouse somatic cells of F0 generation, wherein F0-01 to F0-14 are mouse numbers, WT is a wild type control, M is Marker, H₂O is water control.

FIG. 10: the genotype identification result of mouse somatic cells of F1 generation, wherein 2F 1-01-2F 1-11 is mouse number, PC is positive control, WT is wild control, M is Marker, H is₂O is water control.

FIG. 11: southern blot assay results, 2F1-01 to 2F1-11 are mouse numbers, WT is wild type control.

FIG. 12: CD73 gene humanized mouse CD73 gene schematic two (not to scale).

FIG. 13: the targeting strategy used to prepare humanized mice with the CD73 gene is schematically illustrated in figure three (not to scale).

FIG. 14: the genotype identification result of somatic cells of F1 mouse, wherein 3F1-01 to 3F1-11 are mouse numbers, M is Marker, H₂O is water control;

FIG. 15: the result of RT-PCR is shown schematically, wherein, +/+ is wild type control, H/H is humanized homozygous mouse with CD73 gene, H₂O is water control;

FIG. 16: flow detection results of leukocyte subgroup ratios in the spleen;

FIG. 17: flow assay results for T cell subpopulation ratios in spleen;

FIG. 18: flow detection results of the proportion of leukocyte subsets in lymph nodes;

FIG. 19: flow detection results of the proportion of the T cell subset in the lymph nodes;

FIG. 20: flow detection results of leukocyte subset ratio in peripheral blood;

FIG. 21: flow detection results of the proportion of T cell subsets in peripheral blood;

FIG. 22: blood routine detection results;

FIG. 23: and (5) biochemical detection results of blood.

Detailed Description

The invention will be further described with reference to specific embodiments, and the advantages and features of the invention will become apparent as the description proceeds. These examples are illustrative only and do not limit the scope of the present invention in any way. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention, and that such changes and modifications may be made without departing from the spirit and scope of the invention.

In each of the following examples, the equipment and materials were obtained from several companies as indicated below:

both the C57BL/6 mouse and the Flp tool mouse were purchased from the national rodent laboratory animal seed center of the Chinese food and drug assay institute;

the AIO kit is from Beijing Baiosaixi map gene biotechnology limited company with the cargo number BCG-DX-004;

APC anti-mouse CD73 Antibody (mCD73 APC) was purchased from Biolegend under the accession number 127209;

PerCP/Cy55 Anti-mouse TCR Beta Chain Antibody (mTcR. Beta. PerCP) was purchased from Biolegend under the accession number 109228;

PE anti-human CD73(Ecto-5' -nucleotidase) Antibody (hCD73 PE) was purchased from Bioleged under the code 344003;

FITC anti-mouse CD19 Antibody (hCD19 FITC) from Biolegend under the cat No. 115506;

PE anti-human CD73 Antibody (hCD73 PE) was purchased from Biolegend under the accession number 344003;

PE/Cy^TM7anti-mouse/rat Foxp3 Antibody (mFoxp3 PE) available from eBioscience under a cat number of 25-5773-82;

Brilliant Violet 510^TManti-mouse CD4 Antibody (mCD4) from Biolegend under the code 100559;

APC anti-mouse CD73 Antibody (mCD73 APC) was purchased from Biolegend under the accession number 127209.

The present invention contemplates the modification of a non-human animal, such as a mouse, to include all or part of the gene sequence encoding human CD73 protein in order to express human or humanized CD73 protein in vivo. Non-human mammals expressing human CD73 protein can be obtained by various gene editing systems and preparation methods, including but not limited to gene targeting technology using embryonic stem cells (ES), Zinc Finger Nuclease (ZFN) technology, transcription activator-like effector nuclease (TALEN) technology, homing endonuclease (megabase megaribozyme), Regularly Clustered partitioned Short Palindromic Repeats (CRISPR) technology, or other molecular biology technologies. The embodiment of the invention mainly takes the gene homologous recombination technology of ES cells as an example, and explains how to obtain the humanized mouse by constructing a specific targeting mouse CD73 gene targeting vector to realize sequence replacement.

Example 1 humanized mouse with CD73 Gene

A schematic comparison of the mouse CD73 Gene (NCBI Gene ID: 23959, Primary source: MGI:99782, UniProt ID: Q61503, transcript-based NM-011851.4 (SEQ ID NO:1) and its encoded protein NP-035981.1 (SEQ ID NO:2) and the human CD73 Gene (NCBI Gene ID: 4907, Primary source: HGNC:8021, UniProt ID: P21589, transcript-based NM-002526.4 (SEQ ID NO:3) and its encoded protein NP-002517.1 (SEQ ID NO:4)) is shown in FIG. 1.

Given that human or mouse CD73 has multiple subtypes or transcripts, the methods described herein can be applied to other subtypes or transcripts.

For the purposes of the present invention, the gene sequence encoding the human CD73 protein may be introduced at the endogenous mouse CD73 locus, so that the mouse expresses the human or humanized CD73 protein. Mouse cells are modified by gene editing techniques, such as insertion of a sequence encoding human CD73 protein at exon 2 of endogenous mouse CD73 locus. In order to make the expression of human CD73 protein more stable and better in mice, a mouse promoter is reserved, and P2A (shown as SEQ ID NO: 10) is inserted before the coding sequence of human CD 73. The schematic diagram of the finally obtained transformed humanized mouse CD73 gene is shown in figure 2, the mouse regulates the expression of a humanized CD73 sequence through an endogenous promoter, and the CD73 protein expressed in vivo is a human CD73 protein; and further designs a targeting strategy as shown in fig. 3. Because the transcription termination sequence such as stop codon exists after the inserted recombination sequence, and the partial sequence of the coding region of the mouse is deleted, the mouse coding region on the humanized mouse CD73 gene can not be normally transcribed and translated.

As shown in FIG. 3, the targeting strategy is schematically shown with the upstream and downstream homology arm sequences of mouse CD73, and an A-1 fragment (SEQ ID NO:13) comprising the P2A, human CD73, mouse 3' UTR and STOP sequences on the targeting vector. Wherein the upstream homology arm sequence (5 'homology arm, SEQ ID NO:7) is identical to the nucleotide sequence at positions 88348101 and 88352454 of NCBI accession No. NC-000075.6, and the downstream homology arm sequence (3' homology arm, SEQ ID NO:8) is identical to the nucleotide sequence at positions 88354132 and 88358698 of NCBI accession No. NC-000075.6; the sequence of human CD73 (SEQ ID NO:9) is identical to the nucleotide sequence at positions 58-1782 of NCBI accession No. NM-002526.4; the STOP sequence is shown in SEQ ID NO: 20, the sequence of mouse 3' UTR is shown as SEQ ID NO: 22, respectively. Also included on the A-1 fragment is that used for positive clone screeningThe resistance gene, i.e. neomycin phosphotransferase coding sequence Neo, is provided with two site-specific recombination system Frt recombination sites which are arranged in the same direction on two sides of the resistance gene to form a Neo cassette (Neo cassette). Wherein, the P2A, the human CD73 sequence, the mouse 3 ' UTR and STOP sequence and the Neo cassette sequence are arranged in the A-1 segment in the direction from 5' to 3 ', and the sequence containing the human CD73 gene is directly connected with the upstream homology arm sequence; ligation of the downstream Neo cassette to the murine locus was designed to be 5' -GCGGAACCCTTCGAAGTTCCTATTCTCTAGAAAGTATAGGAACTTCcc agagcccaaggcactcattattagcactgggtgagctccatg-3' (SEQ ID NO:23), wherein the sequence "CTTC"the second" C "of the sequence is the last nucleotide of the Neo cassette"ccagaThe first "c" of "is the first nucleotide of the murine sequence.

In addition, a coding gene with a negative selection marker (diphtheria toxin a subunit coding gene (DTA)) was constructed downstream of the 3' homology arm of the targeting vector.

The construction of the targeting vector can be carried out by adopting a conventional method, such as enzyme digestion connection and the like. The constructed targeting vector can be preliminarily verified by enzyme digestion and then sent to a sequencing company for sequencing verification. The method comprises the steps of performing electroporation transfection on a targeting vector which is verified to be correct by sequencing into embryonic stem cells of a C57BL/6 mouse, screening the obtained cells by using a positive clone screening marker gene, detecting by using PCR and Southern Blot technology to confirm the integration condition of an exogenous gene, screening correct positive clone cells, performing Southern Blot detection on clones which are identified to be positive by PCR, wherein the PCR detection result is shown in figure 4, the result shows that 18 clones are positive clones, and the results show that ZL004, ZL007, 009, ZL010, ZL011, ZL012, ZL013, 014ZL, ZL016 and ZL018 are positive clones and have no random insertion.

The PCR assay included the following primers:

Mut F1：5’-GAGTGGTATGTGCAGCTCAGGGAAG-3’(SEQ ID NO:24)，

Mut R1：5’-GCTCCAAGAAACGAAAAGGCCGAGA-3’(SEQ ID NO:25)；

the selected correct positive clone is introduced into the separated blastocyst (white mouse) according to the known technology in the field, the obtained chimeric blastocyst is transferred into the culture solution for short-term culture and then transplanted into the oviduct of the recipient mother mouse (white mouse), and F0 generation chimeric mouse (black and white alternate) can be produced. The F1 generation of mice are obtained by backcrossing the F0 generation chimeric mice and the wild mice, the Mut F1 primer and the Mut R1 primer are used for detection, the detection result is shown in figure 5, 11F 1 generation mice are all positive mice, the positive mice are obtained, F1 mice are obtained, and the F1 generation heterozygous mice are mutually mated, so that the F2 generation homozygous mice can be obtained. The positive mice can also be mated with Flp tool mice to remove the positive clone selection marker gene (the process is schematically shown in figure 6), and then mated with each other to obtain the humanized homozygote mice (V1) expressing the CD73 gene of the human CD73 protein.

Humanized mice can also be obtained using other gene editing methods, such as gene editing using the CRISPR/Cas system, designing the targeting strategy as shown in fig. 7, in which the targeting vector is shown in fig. 7 to contain the homology arm sequences upstream and downstream of the mouse CD73 gene, and an a-2 fragment comprising the nucleotide sequence of human CD73, wherein the upstream homology arm sequence (5 'homology arm, SEQ ID NO: 5) is identical to the nucleotide sequence at position 88350685 and 88352454 of NCBI accession No. NC _000075.6, and the downstream homology arm sequence (3' homology arm, SEQ ID NO: 6) is identical to the nucleotide sequence at position 88353811 and 88354943 of NCBI accession No. NC _ 000075.6; wherein, the nucleotide sequence of the A-2 fragment is shown as SEQ ID NO. 12, the nucleotide sequence of the human CD73 is shown as SEQ ID NO. 9, and the nucleotide sequence is the same as the nucleotide sequence of the A-1 fragment human CD73 in the picture 3. The protein sequences of the reconstructed humanized mice are respectively SEQ ID NO. 4.

The construction of the targeting vector can be carried out by adopting a conventional method, such as enzyme digestion connection and the like. And carrying out preliminary verification on the constructed targeting vector by enzyme digestion, and then sending the targeting vector to a sequencing company for sequencing verification. The correct targeting vector was verified by sequencing for subsequent experiments.

The target sequence determines the targeting specificity of the sgRNA and the efficiency of inducing Cas9 to cleave the gene of interest. Therefore, efficient and specific target sequence selection and design are a prerequisite for constructing sgRNA expression vectors. sgRNA sequences that recognize the target site were designed and synthesized. The target sites are located on exon 2 and intron 2-3 of the CD73 gene, and the target site sequence of each sgRNA on CD73 is as follows:

SgRNA1 target site sequence (SEQ ID NO: 26): 5'-AGCGCAAACATTAAGGCACGGGG-3', respectively;

SgRNA2 target site sequence (SEQ ID NO: 27): 5'-TTAATGTTTGCGCTCAGAATTGG-3', respectively;

SgRNA3 target site sequence (SEQ ID NO: 28): 5'-GAGCGCAAACATTAAGGCACGGG-3', respectively;

SgRNA4 target site sequence (SEQ ID NO: 29): 5'-AATTCTGAGCGCAAACATTAAGG-3', respectively;

SgRNA5 target site sequence (SEQ ID NO: 30): 5'-CCTCTAGCACATCAGATATCTGG-3', respectively;

SgRNA6 target site sequence (SEQ ID NO: 31): 5'-TGAGCGCAAACATTAAGGCACGG-3', respectively;

SgRNA7 target site sequence (SEQ ID NO: 32): 5'-GATTGTTGGATATACTTCAAAGG-3', respectively;

SgRNA8 target site sequence (SEQ ID NO: 33): 5'-TTGGAAATTTAACGTTTCTGAGG-3', respectively;

SgRNA9 target site sequence (SEQ ID NO: 34): 5'-CATCACTTGCGGGGAAGTCCTGG-3', respectively;

SgRNA10 target site sequence (SEQ ID NO: 35): 5'-TGCTGAGTCCATCACTTGCGGGG-3', respectively;

SgRNA11 target site sequence (SEQ ID NO: 36): 5'-CAGGACTTCCCCGCAAGTGATGG-3', respectively;

SgRNA12 target site sequence (SEQ ID NO: 37): 5'-GTCTGTAACCCCGATAAGCTTGG-3', respectively;

SgRNA13 target site sequence (SEQ ID NO: 38): 5'-TTGCATCTGCGGAGTTAGCATGG-3', respectively;

SgRNA14 target site sequence (SEQ ID NO: 39): 5'-GCCATCGACCATTCGTGCACAGG-3', respectively;

SgRNA15 target site sequence (SEQ ID NO: 40): 5'-CCCTGTGCACGAATGGTCGATGG-3', respectively;

SgRNA16 target site sequence (SEQ ID NO: 41): 5'-AGTGGAGCCCTGTGCACGAATGG-3', respectively;

the activity of multiple sgrnas is detected by using a UCA kit, and the sgrnas have different activities as shown in the results, and the detection results are shown in table 1 and fig. 8.

Table 1: relative activity detection result of sgRNA

Although the sgRNA2, sgRNA 4and sgRNA8 are relatively low in activity, which may be caused by specificity of a target site sequence, according to our experiments, the values of sgRNA2, sgRNA 4and sgRNA8 are still significantly higher than those of a control group, and it can still be judged that sgRNA2, sgRNA 4and sgRNA8 are active, and the activity meets the requirements of gene targeting experiments, and sgRNA7 and sgRNA14 are selected for subsequent experiments. The 5' end and the complementary strand are respectively added with enzyme cutting sites to obtain a forward oligonucleotide and a reverse oligonucleotide (see table 2), and after annealing, the annealing products are respectively connected to pT7-sgRNA plasmids (the plasmids are firstly linearized by BbsI), so as to obtain expression vectors pT7-CD73-7 and pT7-CD 73-14.

Table 2 sgRNA7 and sgRNA14 forward and reverse oligonucleotide sequences

pT7-sgRNA vector was synthesized by plasmid synthesis company as a fragment DNA (SEQ ID NO: 21) containing the T7 promoter and sgRNA scaffold, and ligated to a backbone vector (Takara, cat. No. 3299) by enzyme digestion (EcoRI and BamHI) in sequence, and sequencing by the professional sequencing company was verified, and the result indicated that the objective plasmid was obtained.

Taking wild type C57BL/6 mouse fertilized eggs, and injecting in vitro transcription products of pT7-CD73-7 and pT7-CD73-14 plasmids (transcription is carried out by using an Ambion in vitro transcription kit according to the instruction method), a targeting vector and Ca s9 mRNA into cytoplasm or nucleus of the mouse fertilized eggs after being premixed by using a microinjection instrument. Microinjection of fertilized eggs was performed according to the method in the manual of experimental manipulation of mouse embryos (third edition), published by chemical industry, 2006, and the injected fertilized eggs were transferred to a culture medium for short-term culture and then transplanted into the oviduct of a recipient mother mouse for development, and the obtained mice (generation F0) were subjected to hybridization and selfing to expand the population number and establish stable CD73 humanized mice.

The genotype of somatic cells of F0 mouse can be identified by the same PCR method as that described in the example (the primer sequences are the same), and the identification results of some F0 mice are shown in FIG. 9, in which 5 mice numbered F0-01, F0-02, F0-04, F0-11 and F0-13 are positive mice, and the 8 mice are further verified to be positive mice without random insertion by sequencing.

WT-F：5’-TTTGATAACGGTGTGGAAGGACTGAT-3’(SEQ ID NO：48)

WT-R：5’-AGACAGCGATGCCCTTCAGAGGATA-3’(SEQ ID NO：49)

WT-F：5’-TTTGATAACGGTGTGGAAGGACTGAT-3’(SEQ ID NO：48)

Mut-R：5’-TACACGGTGAACCAGATAGTGCCC-3’(SEQ ID NO：50)

GT-F：5’-GTGGGCGGAATCCATGTGGTGTATG-3’(SEQ ID NO：51)

GT-R：5’-GCCCCAGGGCGATGATCTTATTCAC-3’(SEQ ID NO：52)

CD73 humanized mice identified as positive for F0 were mated with C57BL/6 wild-type mice to give F1 generation mice. The same PCR method was used to genotype the F1 generation mice, and the results are shown in FIG. 10, which shows that 11 mice numbered 2F1-01 to 2F1-11 were all positive mice. The 11 mice identified as positive by F1 PCR were subjected to Southern blot analysis to confirm the presence of random insertions. Cutting rat tail to extract genome DNA, digesting genome with BglII enzyme or NcoI enzyme, transferring membrane and hybridizing. Probes 5 'Probe and 3' Probe are located on the left side of the 5 'homology arm and on the 3' homology arm, respectively, and the lengths of the probes and the target fragment are shown in Table 3.

TABLE 3 lengths of the particular probes and target fragments

Restriction enzyme	Probe needle	Wild type fragment size	Recombinant sequence fragment size
				BglII	5’Probe	3.1kb	5.2kb
NcoI	lox2 Stop Probe(3’)	--	4.6kb

The probe synthesis primers were as follows:

5’Probe-F(SEQ ID NO：53)：5’-AGAGACCCTTTAGACCTGGATCAGC-3’

5’Probe-R(SEQ ID NO：54)：5’-ACCTAGAAAACTTTGGTGGGACATGA-3’

lox2 Stop Probe(3’)-F(SEQ ID NO：55)：5’-AACTGATGAATGGGAGCAGTGGTGG-3’

lox2 Stop Probe(3’)-R(SEQ ID NO：56)：5’-GCAGACACTCTATGCCTGTGTGGAG-3’

the Southern blot assay results are shown in FIG. 11. The results of 5' Probe and STOP Probe were combined and further sequence verified that 11 mice numbered 2F1-01 to 2F1-11 had no random insertions, confirming that these 11 mice were positive heterozygous mice and had no random insertions. This shows that using this method, a humanized genetically engineered mouse of CD73 (V1) can be constructed that can be passaged stably and without random insertions.

RT-PCR can be used to detect the expression of humanized CD73mRNA in humanized mice with the CD73 gene. 1 mouse of 7-week-old wild type C57BL/6 and humanized CD73 homozygote mouse are respectively selected, bladder tissues are taken after neck-removing euthanasia, total RNA of cells is extracted, and PCR amplification is carried out after reverse transcription is carried out to cDNA by utilizing a reverse transcription kit, wherein the primer sequences are shown in Table 4.

TABLE 4 RT-PCR detection primer sequences and target fragment lengths

The test results showed (see FIG. 15) that murine CD73mRNA expression was detectable in wild type C57BL/6 mouse cells and humanized CD73mRNA expression was not detected; humanized CD73mRNA expression was detected in humanized CD73 homozygous mouse cells, and murine CD73mRNA expression was not detected.

In order to obtain the CD73 humanized mouse, the human CD73 genomic sequence may be replaced on the mouse locus, the transformation of the mouse CD73 gene is realized by the BAC transgenic method, the schematic diagram of the humanized mouse CD73 gene finally obtained is shown in fig. 12, and the targeting strategy diagram shown in fig. 13 is designed. The figure shows the A-3 fragment containing the sequences of the upstream and downstream homology arms of mouse CD73, and the human CD73 gene on the targeting vector. Wherein the upstream homology arm sequence (5 'homology arm, SEQ ID NO:15) is identical to the nucleotide sequence at positions 88323954 and 88327685 of NCBI accession No. NC-000075.6, and the downstream homology arm sequence (3' homology arm, SEQ ID NO:16) is identical to the nucleotide sequence at positions 88372532 and 88377482 of NCBI accession No. NC-000075.6; the sequence of human CD73 (SEQ ID NO:17) is identical to the nucleotide sequence at positions 85450140-85496067 of NCBI accession number NC-000006.12; the A-3 fragment (shown as SEQ ID NO: 18) also comprises a resistance gene used for positive clone screening, namely neomycin phosphotransferase coding sequence Neo, and two site-specific recombination system LOXP recombination sites which are arranged in the same direction are arranged on two sides of the resistance gene.

The construction of the targeting vector can be carried out by adopting a conventional method, such as enzyme digestion connection and the like. The constructed targeting vector can be preliminarily verified by enzyme digestion and then sent to a sequencing company for sequencing verification. The method comprises the steps of transfecting a targeting vector with correct sequencing verification into embryonic stem cells of a C57BL/6 mouse by means of electroporation, screening the obtained cells by means of positive clone screening marker genes, detecting and confirming the integration condition of exogenous genes by means of PCR and Southern Blot technologies, screening correct positive clone cells, carrying out Southern Blot detection on clones identified as positive by PCR, sending sequencing confirmation, and selecting the positive clones without random insertion clones for the next step.

The PCR assay included the following primers:

L-GT-F：5’-AAGATGTTTCTTGTCCCGGGTTTCG-3’(SEQ ID NO：57)

L-GT-R：5’-GCAGTAAATGAGAGCTGCGTGAGGG-3’(SEQ ID NO：14)

the selected correct positive clone is introduced into the separated blastocyst (white mouse) according to the known technology in the field, the obtained chimeric blastocyst is transferred into the culture solution for short-term culture and then transplanted into the oviduct of the recipient mother mouse (white mouse), and F0 generation chimeric mouse (black and white alternate) can be produced. The F1 generation mice are obtained by backcrossing the F0 generation chimeric mice and the wild mice, the detection result is shown in figure 14 by using the L-GT-F and L-GT-R primers, and the result shows that 11 mice are all positive mice. And mating the F1 generation heterozygous mice to obtain F2 generation homozygous mice. The positive mice and Cre tool mice can be mated to remove the positive clone screening marker gene, and then the positive mice and the Cre tool mice are mated with each other to obtain the CD73 gene humanized homozygote mice (V2) expressing the human CD73 protein. The coding region and the protein sequence of the mRNA sequence of the humanized mouse after being transformed are respectively SEQ ID NO 19 and SEQ ID NO 4.

Example 2 humanized protein expression of CD73 Gene humanized mice in vivo

The expression of the humanized CD73 protein in mice was confirmed by a conventional assay method, and in this example, two types of humanized CD73 genes obtained in example 1 were assayed for protein expression by flow cytometry.

8 weeks old wild type C57BL/6 mice, and V1 and V2 CD73 gene humanized homozygote mice were selected 1 each, splenocytes were harvested after euthanasia, and anti-mouse TCR β Antibody PerCP/Cy5.5 anti-mouse TCR β chain Antibody (mTcR β PerCP), anti-mouse CD19 Antibody FITC anti-mouse CD19 Antibody (hCD19 FITC), anti-human CD73 Antibody PE anti-human CD73 Antibody (hCD73 PE), anti-mouse Foxp3 Antibody PE/CyTM7 anti-mouse/rate Foxp3 Antibody (mFoxp3 PE), anti-mouse CD4 Antibody Brilliant Violet 510 Antibody^TMFlow detection was performed after anti-mouse CD4 Antibody (mCD4) and anti-mouse CD73 Antibody APC anti-mouse CD73(mCD73 APC) recognition staining, and the results are shown below:

TABLE 5 flow assay results

The results show that the mouse CD73 protein is detected in a C57BL/6 mouse, and the human CD73 protein is not detected; human CD73 protein was detected in humanized homozygous mice of CD73 gene V1 version and V2 version, and no murine CD73 protein was detected.

The white blood cell and T cell immunotyping in spleen, lymph node and peripheral blood of wild type C57BL/6 mice and V1 version CD73 humanized homozygote mice were further examined by flow cytometry. The results of the measurement of the leukocyte subtypes and T-cell subtypes in the spleen are shown in FIGS. 16 and 17, respectively, and it can be seen from the graphs that the leukocyte subtypes such as B cells, T cells, NK cells, CD4+ T cells, CD8+ T cells, granulocytes (Granulocyte), DC cells, macrophages (Macrophage) and monocytes (Monocyte) in the spleen sample of the humanized homozygous mouse with the CD73 gene are substantially identical to those of the wild-type C57BL/6 mouse (FIG. 16), and the percentages of the T-cell subtypes such as CD4+ T cells, CD8+ T cells and Tregs cells are identical to those of the wild-type C57BL/6 mouse (FIG. 17).

The results of detecting the leukocyte subtypes and the T-cell subtypes in the lymph nodes are shown in FIG. 18 and FIG. 19, respectively, and it can be seen from the graphs that the leukocyte subtypes such as B cells, T cells, NK cells, CD4+ T cells, CD8+ T cells and the like in the lymph node sample of the humanized homozygote mouse with the CD73 gene are consistent with those of the wild-type mouse with C57BL/6 (FIG. 18), and the percentages of the T-cell subtypes such as CD4+ T cells, CD8+ T cells and Tregs cells are substantially consistent with those of the wild-type mouse with C57BL/6 (FIG. 19).

The results of the detection of the leukocyte subtypes and the T-cell subtypes in the peripheral blood are shown in FIGS. 20 and 21, respectively, and it can be seen from the graphs that the leukocyte subtypes such as B-cells, T-cells, NK-cells, CD4+ T-cells, CD8+ T-cells, granulocytes (Granulocyte), DC-cells, macrophages (Macrophage) and monocytes (Monocyte) in the peripheral blood sample of the humanized homozygous mouse with the CD73 gene are substantially identical to those of the wild-type C57BL/6 mouse (FIG. 20), and the percentage of the T-cell subtypes such as CD4+ T-cells, CD8+ T-cells and Tregs-cells are substantially identical to those of the wild-type C57BL/6 mouse (FIG. 21).

The above results indicate that the humanization of the CD73 gene did not affect the differentiation, development and distribution of leukocytes and T cells in mice in spleen, lymphoid tissue and peripheral blood.

Further, the wild-type C57BL/6 mouse and the CD73 humanized homozygous mouse were separately collected and subjected to blood routine and blood biochemical tests, the blood routine results are shown in fig. 22, and the results of blood routine are substantially identical to those of the C57BL/6 wild-type mouse in terms of white blood cell count (WBC), red blood cell count (RBC), hemoglobin concentration (HB), Hematocrit (HCT), Mean Corpuscular Volume (MCV), mean corpuscular hemoglobin content (MCH), Mean Corpuscular Hemoglobin Concentration (MCHC), red blood cell distribution width (RDW), platelet count (PLT), Mean Platelet Volume (MPV), neutrophil count (NE), lymphocyte count (LY), and monocyte count (MO) in the peripheral blood sample of the CD73 gene humanized homozygous mouse; the biochemical blood test results are shown in fig. 23, and the results of alanine Aminotransferase (ALT), aspartate Aminotransferase (AST), serum Total Cholesterol (TC), Creatinine (CR), blood Glucose (GLU), Triglyceride (TRIG), UREA (UREA) and serum Albumin (ALB) in the peripheral blood sample of the humanized homozygote mouse with the CD73 gene are substantially the same as those of the wild-type mouse with C57BL/6 gene. The above results indicate that the humanized modification of the CD73 gene did not affect the composition and morphology of blood cells, nor did it alter AST and ALT levels and liver health status.

Example 3 Dual-or Multi-Gene-humanized mice

A multiple humanized mouse model containing a humanized CD73 gene can also be prepared using the previously prepared humanized mouse of CD73 gene. For example, fertilized egg cells used in microinjection and embryo transfer processes are selected from fertilized egg cells derived from other genetically modified mice, and for example, a two-gene humanized mouse model of PD-1 and CD73 can be obtained by gene editing of fertilized egg cells of a PD-1 humanized mouse. The CD73 humanized mouse homozygote or heterozygote can also be mated with other gene modified homozygote or heterozygote mice, the offspring thereof is screened, according to Mendelian inheritance rule, the CD73 humanized mouse and other gene modified double-gene or multi-gene modified heterozygote mice can be obtained with a certain probability, and then the heterozygotes are mated with each other to obtain double-gene or multi-gene modified homozygotes.

Taking a double humanized CTLA4/CD73 mouse as an example, because the CTLA 4and the CD73 genes of the mouse are located on chromosome 1 and chromosome 9 respectively, a CTLA4 humanized mouse and a CD73 humanized mouse are selected to mate, and a positive progeny mouse is screened to obtain a double humanized CTLA4/CD73 gene mouse.

Example 4 evaluation of drug efficacy

The humanized mouse with the CD73 gene prepared by the method can be used for evaluating the drug effect of the regulator targeting human CD 73. The CD73 humanized mouse homozygote is inoculated subcutaneously with mouse colon cancer cell MC38-hCD73 (human CD73 expressing MC38 cell) until the tumor volume grows to about 100mm³Then dividing into control group or treatment group (n is 5) according to tumor volume, injecting 10mg/kg anti-human CD73 antibody MEDI9447 into abdominal cavity of treatment group, injecting equal volume hIgG into control group, wherein the administration frequency is 2 times per week and 6 times in total, and the tumor volume of single mouse after inoculation reaches 3000mm³An euthanasia end test was performed. Tumor volumes were measured periodically and the mice were weighed. On the whole, the health status of the animals in the treatment group and the control group was good during the experiment. At the end of the experiment, there was an increase in the weight average of the animals in each group, and there was no significant difference in the body weight of mice in the treated and control groups throughout the experimental period, indicating that the animals were well tolerated MEDI 9447. However, from the results of tumor measurement, at the end of the experiment, the treatment groupThe tumor volume of the mice was significantly smaller than the control group, and it was seen that the tumor growth in the mice was significantly inhibited after treatment with MEDI9447, a human CD73 antibody. The research results show that the anti-human CD73 antibody has obvious inhibition effect on the tumor growth in a CD73 humanized mouse, so that the humanized CD73 animal model can be used for evaluating the effectiveness of a targeted CD73 medicament and evaluating the treatment effect of the targeted CD73 in vivo.

The preferred embodiments of the present invention have been described in detail, however, the present invention is not limited to the specific details of the above embodiments, and various simple modifications may be made to the technical solution of the present invention within the technical idea of the present invention, and these simple modifications are within the protective scope of the present invention.

It should be noted that the various technical features described in the above embodiments can be combined in any suitable manner without contradiction, and the invention is not described in any way for the possible combinations in order to avoid unnecessary repetition.

In addition, any combination of the various embodiments of the present invention is also possible, and the same should be considered as the disclosure of the present invention as long as it does not depart from the spirit of the present invention.

Sequence listing

<110> Baiosai Diagram (Beijing) pharmaceutical science and technology Co., Ltd

Construction method and application of <120> CD73 gene humanized non-human animal

<130> 1

<150> CN202010807504.X

<151> 2020-08-12

<160> 63

<170> SIPOSequenceListing 1.0

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agtttagtag aggccccggt ggccggtgct cactgggctc agggccagtc cacccgctca 60

acgcgctcta gccagccatg cgtcccgcgg ccgctaaggt acccaagtgg ctgcttctcg 120

cactgagcgc tctactacca cagtggcccg cagccagtgc ctgggagctc acgatcctgc 180

acacaaacga cgtgcacagc cggctagagc agaccagcga tgactccacc aagtgcctca 240

acgccagcct gtgtgtgggc ggcgtggccc ggctctttac caaggtgcag cagatccgca 300

aggaagaacc caacgtgctg tttttggatg ccggagacca gtaccagggc accatctggt 360

tcaccgttta caaaggcctt gaagtggcac acttcatgaa catcctgggc tacgatgcta 420

tggcactggg aaatcatgaa tttgataacg gtgtggaagg actgattgat cccctcctca 480

gaaacgttaa atttccaatt ctgagcgcaa acattaaggc acgggggcct ctagcacatc 540

agatatctgg actttttctg ccatctaaag ttctctctgt tggcggtgag gttgtgggga 600

ttgttggata tacttcaaag gaaacccctt tcctctcaaa tccagggaca aatttagtct 660

ttgaagatga aatctctgca ttgcagcctg aagtagataa actaaagacg ctaaatgtga 720

ataagatcat cgccctgggg cactctggtt ttgagatgga caaacttatc gctcagaaag 780

ttcgaggtgt ggacatcgtg gtgggaggac actccaacac ctttctctac acaggaaatc 840

caccttccaa agaagtgcct gcggggaagt acccattcat agtcaccgca gatgatggac 900

ggcaggtgcc tgtggtccag gcctatgcct ttggcaaata cctgggctac ctgaaggttg 960

agtttgatga taaaggcaat gttatcactt cctatggaaa tcccattctt ctcaacagca 1020

gcattcctga agatgcgacc atcaaagcag acattaacca atggaggata aaattagata 1080

attattctac ccaggaactc gggagaacga tcgtctacct ggatggctcc actcagacgt 1140

gccgcttcag ggaatgcaac atgggaaacc tgatctgtga tgccatgatt aacaacaacc 1200

tcagacaccc agatgaaatg ttttggaacc acgtgtccat gtgcattgta aacggaggtg 1260

gcatccggtc ccccattgat gagaagaaca atggtaccat cacctgggag aacctggctg 1320

ctgtgctgcc ctttggaggg acatttgacc tcgtccaatt aaaagggtcc accctgaaga 1380

aggcttttga gcacagcgtg catcgctatg gccagtccac cggagagttc ctgcaagtgg 1440

gtggaatcca tgtggtgtac gatattaacc gaaagccctg gaacagagtg gtccaattag 1500

aagttctctg caccaagtgt cgagtgccca tctatgagcc tcttgaaatg gataaagtgt 1560

ataaagtgac cctcccaagc tatctggcca acggtggaga tggattccag atgataaaag 1620

atgaattact aaagcatgac tctggtgatc aagatatcag cgtggtttct gaatacatct 1680

caaaaatgaa agtagtttac ccagccgttg aagggcggat caagttctct gcagcaagtc 1740

attaccaggg aagctttcct ttagtaattc tttcattttg ggcaatgatc cttattttgt 1800

accaataaca gggagtctcc ttgtccttga tgtcaaacta catttttctt ccagtgatat 1860

tcatatctgc ctctggagac ctggctttgt aacagcactc atcatcctca aggttcctag 1920

cagatgttct tcacaaggaa gagactgtaa catcatttgt tggggccagc aactcagtga 1980

gcagatagaa agtcacagtg aaccaacagg gtccttctgg cagggagtgg gtaaggggaa 2040

acaactagat gtagcttgca tagccacata acacatctgg ttaccatttc cctttctatt 2100

catttctaat ccatcaaaca attgatgttt acatacaact tcatcatcgc cagctctggt 2160

ggcacatgcc tgtggtcacg gcacttggca gggaggagag gatggctgca agttctaggc 2220

cagcctgaca tatgtagagt ttcaggccag tcagctagat atcaagactc acacacacaa 2280

acaaacatta taatttacaa gtagattttt gtagacaagt cttatgataa gctaagcaga 2340

aagggtcgac ttgttcaagg tcagacatct ccaaataata catctaagat ctgtacctga 2400

gtctttttac ctcaagtcca acctctcgcc tactgtcaag tctcctctct tcctgcggtt 2460

ggtctcagac aacaaatctg tttctgcctt ccatagtacc ctttcttttg ggctcttgtt 2520

gtctctcagg tttgagagag tagctactgg acaggattct ttcctacaac accatgtgcc 2580

tttggtgagt catagacaaa ctgtatacag cagataatag attagtccag ggactgcaaa 2640

aggcagtcag ggacaggcag gaaagggaaa ggagaatatg accaggactt acagaagaga 2700

gtagatgggt tccataaaca ctagttaagg agcagaacat ggctccaaag cacaacactg 2760

gttcttatta actcctgtgc gactccaaaa gccctgcctc tctccttcct tcttcctcac 2820

atggaggtgc catgagagaa cttcctaaaa taaactgtaa ttctcaacct gcacctgtcc 2880

tgtccaagat gccaacacgg ctctcattca agtcacacat gtacaatcca atattctgga 2940

aggcaagagt gaatttaaga agtggtatac ataaatgttt gaagtcactg agactcaaat 3000

cccacacaac cactgtaagg catactcagg tcaagacatg agaagaccag caggacagtg 3060

agccctatgg gaggcccaag agatacccag actgatgaaa tgatctgtgc ccacagaaca 3120

ccagaccaag acagtcctgt gaccaagtga gcatagtcag cactgtacag tagttaagaa 3180

tccttaacta taaaccagca gctagatgtc tggacaggga acatgttggt ttcctagtgt 3240

ttacgaatat taaggactct tgacccaaac cgtttaaaat tccaatcctt ttatgaagtt 3300

tgttgcagtg aaacaatgcc ttcagttctt tttgaatgtg tagattagtt ataaacagaa 3360

atgtcagtaa gactgtaaaa aaaaatgaag aaaggtgcat atgaggtata tttttatgct 3420

tggcgagtaa ataaggagaa atcttatagc ataattttta aagaacattt gtataatttt 3480

tctaaatatg tgtatatgta tattttatgc agcagtattg agaaatgacc ttggactatc 3540

ttgtaattgt aaattccaaa aataaagctg aagatatagt 3580

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Met Arg Pro Ala Ala Ala Lys Val Pro Lys Trp Leu Leu Leu Ala Leu

1 5 10 15

Ser Ala Leu Leu Pro Gln Trp Pro Ala Ala Ser Ala Trp Glu Leu Thr

20 25 30

Ile Leu His Thr Asn Asp Val His Ser Arg Leu Glu Gln Thr Ser Asp

35 40 45

Asp Ser Thr Lys Cys Leu Asn Ala Ser Leu Cys Val Gly Gly Val Ala

50 55 60

Arg Leu Phe Thr Lys Val Gln Gln Ile Arg Lys Glu Glu Pro Asn Val

65 70 75 80

Leu Phe Leu Asp Ala Gly Asp Gln Tyr Gln Gly Thr Ile Trp Phe Thr

85 90 95

Val Tyr Lys Gly Leu Glu Val Ala His Phe Met Asn Ile Leu Gly Tyr

100 105 110

Asp Ala Met Ala Leu Gly Asn His Glu Phe Asp Asn Gly Val Glu Gly

115 120 125

Leu Ile Asp Pro Leu Leu Arg Asn Val Lys Phe Pro Ile Leu Ser Ala

130 135 140

Asn Ile Lys Ala Arg Gly Pro Leu Ala His Gln Ile Ser Gly Leu Phe

145 150 155 160

Leu Pro Ser Lys Val Leu Ser Val Gly Gly Glu Val Val Gly Ile Val

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Gly Tyr Thr Ser Lys Glu Thr Pro Phe Leu Ser Asn Pro Gly Thr Asn

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Leu Val Phe Glu Asp Glu Ile Ser Ala Leu Gln Pro Glu Val Asp Lys

195 200 205

Leu Lys Thr Leu Asn Val Asn Lys Ile Ile Ala Leu Gly His Ser Gly

210 215 220

Phe Glu Met Asp Lys Leu Ile Ala Gln Lys Val Arg Gly Val Asp Ile

225 230 235 240

Val Val Gly Gly His Ser Asn Thr Phe Leu Tyr Thr Gly Asn Pro Pro

245 250 255

Ser Lys Glu Val Pro Ala Gly Lys Tyr Pro Phe Ile Val Thr Ala Asp

260 265 270

Asp Gly Arg Gln Val Pro Val Val Gln Ala Tyr Ala Phe Gly Lys Tyr

275 280 285

Leu Gly Tyr Leu Lys Val Glu Phe Asp Asp Lys Gly Asn Val Ile Thr

290 295 300

Ser Tyr Gly Asn Pro Ile Leu Leu Asn Ser Ser Ile Pro Glu Asp Ala

305 310 315 320

Thr Ile Lys Ala Asp Ile Asn Gln Trp Arg Ile Lys Leu Asp Asn Tyr

325 330 335

Ser Thr Gln Glu Leu Gly Arg Thr Ile Val Tyr Leu Asp Gly Ser Thr

340 345 350

Gln Thr Cys Arg Phe Arg Glu Cys Asn Met Gly Asn Leu Ile Cys Asp

355 360 365

Ala Met Ile Asn Asn Asn Leu Arg His Pro Asp Glu Met Phe Trp Asn

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His Val Ser Met Cys Ile Val Asn Gly Gly Gly Ile Arg Ser Pro Ile

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Asp Glu Lys Asn Asn Gly Thr Ile Thr Trp Glu Asn Leu Ala Ala Val

405 410 415

Leu Pro Phe Gly Gly Thr Phe Asp Leu Val Gln Leu Lys Gly Ser Thr

420 425 430

Leu Lys Lys Ala Phe Glu His Ser Val His Arg Tyr Gly Gln Ser Thr

435 440 445

Gly Glu Phe Leu Gln Val Gly Gly Ile His Val Val Tyr Asp Ile Asn

450 455 460

Arg Lys Pro Trp Asn Arg Val Val Gln Leu Glu Val Leu Cys Thr Lys

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Cys Arg Val Pro Ile Tyr Glu Pro Leu Glu Met Asp Lys Val Tyr Lys

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Val Thr Leu Pro Ser Tyr Leu Ala Asn Gly Gly Asp Gly Phe Gln Met

500 505 510

Ile Lys Asp Glu Leu Leu Lys His Asp Ser Gly Asp Gln Asp Ile Ser

515 520 525

Val Val Ser Glu Tyr Ile Ser Lys Met Lys Val Val Tyr Pro Ala Val

530 535 540

Glu Gly Arg Ile Lys Phe Ser Ala Ala Ser His Tyr Gln Gly Ser Phe

545 550 555 560

Pro Leu Val Ile Leu Ser Phe Trp Ala Met Ile Leu Ile Leu Tyr Gln

565 570 575

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actcgccggc actcgcccgg ctcgcccgct ttcgcaccca gttcacgcgc cacagctatg 60

tgtccccgag ccgcgcgggc gcccgcgacg ctactcctcg ccctgggcgc ggtgctgtgg 120

cctgcggctg gcgcctggga gcttacgatt ttgcacacca acgacgtgca cagccggctg 180

gagcagacca gcgaggactc cagcaagtgc gtcaacgcca gccgctgcat gggtggcgtg 240

gctcggctct tcaccaaggt tcagcagatc cgccgcgccg aacccaacgt gctgctgctg 300

gacgccggcg accagtacca gggcactatc tggttcaccg tgtacaaggg cgccgaggtg 360

gcgcacttca tgaacgccct gcgctacgat gccatggcac tgggaaatca tgaatttgat 420

aatggtgtgg aaggactgat cgagccactc ctcaaagagg ccaaatttcc aattctgagt 480

gcaaacatta aagcaaaggg gccactagca tctcaaatat caggacttta tttgccatat 540

aaagttcttc ctgttggtga tgaagttgtg ggaatcgttg gatacacttc caaagaaacc 600

ccttttctct caaatccagg gacaaattta gtgtttgaag atgaaatcac tgcattacaa 660

cctgaagtag ataagttaaa aactctaaat gtgaacaaaa ttattgcact gggacattcg 720

ggttttgaaa tggataaact catcgctcag aaagtgaggg gtgtggacgt cgtggtggga 780

ggacactcca acacatttct ttacacaggc aatccacctt ccaaagaggt gcctgctggg 840

aagtacccat tcatagtcac ttctgatgat gggcggaagg ttcctgtagt ccaggcctat 900

gcttttggca aatacctagg ctatctgaag atcgagtttg atgaaagagg aaacgtcatc 960

tcttcccatg gaaatcccat tcttctaaac agcagcattc ctgaagatcc aagcataaaa 1020

gcagacatta acaaatggag gataaaattg gataattatt ctacccagga attagggaaa 1080

acaattgtct atctggatgg ctcctctcaa tcatgccgct ttagagaatg caacatgggc 1140

aacctgattt gtgatgcaat gattaacaac aacctgagac acacggatga aatgttctgg 1200

aaccacgtat ccatgtgcat tttaaatgga ggtggtatcc ggtcgcccat tgatgaacgc 1260

aacaatggca caattacctg ggagaacctg gctgctgtat tgccctttgg aggcacattt 1320

gacctagtcc agttaaaagg ttccaccctg aagaaggcct ttgagcatag cgtgcaccgc 1380

tacggccagt ccactggaga gttcctgcag gtgggcggaa tccatgtggt gtatgatctt 1440

tcccgaaaac ctggagacag agtagtcaaa ttagatgttc tttgcaccaa gtgtcgagtg 1500

cccagttatg accctctcaa aatggacgag gtatataagg tgatcctccc aaacttcctg 1560

gccaatggtg gagatgggtt ccagatgata aaagatgaat tattaagaca tgactctggt 1620

gaccaagata tcaacgtggt ttctacatat atctccaaaa tgaaagtaat ttatccagca 1680

gttgaaggtc ggatcaagtt ttccacagga agtcactgcc atggaagctt ttctttaata 1740

tttctttcac tttgggcagt gatctttgtt ttataccaat agccaaaaat tctccttgcc 1800

tttaatgtgt gaaactgcat tttttcaagt gagattcaaa tctgcctttt aggacctggc 1860

tttgtgacag caaaaaccat ctttacaggc tcctagaagc tgaaggttag agcattataa 1920

aatgaagaga cagacatgat tactcagggt cagcaaccta gtgagttaga aaaaaaatta 1980

acatagggcc ctataaggag aaagccaact atgttaagtt tacgtgtcca aattttaatg 2040

aaattttact aacaatttta aaccatattt ttcttcttca tatccatttc taatccatca 2100

aacagcttat gtttacataa aattttatca ttcacaagga agttttaagc acactgtctc 2160

atttgatatc cacaacttat ttttggtagg aaagagagat gtttttccca cctgtcagat 2220

gaaaaaactg aagctcaaaa agggttgact tgaccataca gctaatgctg acagatccaa 2280

gacctagacc taggtctttt gaactcaagt ccagcattct caactatatc aagttactgt 2340

tcagaatact taatatctcc tctcttcata attatcaata gccccaagct catggatgac 2400

aaatctctgc tttatttctt gtctctattt tttcacttta tagctcctgt tataatagca 2460

agtttaatgg tataaacaca ggataccatc ctctcttgca acacccatgt gcctttgatg 2520

agtcaggtag caagctgtag tagataatga gaaaggccag aggctgcaaa agacagtcaa 2580

aggacacgag agaaaggaag gggaagaaca ggactccagg actgttttat attatagaaa 2640

agcaagagct aaagagcatt tacacatgtt aaacagatac ttgttaagca tagtgcctga 2700

cacacggcat tagctgttat tttatgagat tccatcagct ctgcctctgt cctctttctt 2760

ctaacatgaa ggtatcatga gaagagaacc ttctaacata agctgtaatt ctaaacctgc 2820

acttgtccct ctccagcaag aggctagcac tgaattcatt ctactcatac tacacaccca 2880

gttatggaat gtccagagtt ctcgaagaaa ataaatgact ttaggaagag gtatacattt 2940

tttaagtcgc tctgcctcca aatctgaaca gtcactgtaa atcattctta agcccagata 3000

tgagaacttc tgctggaaag tgggaccctc tgagtgggtg gtcagaaaat acccatgctg 3060

atgaaatgac ctatgcccaa agaacaaata cttaacgtgg gagtggaacc acatgagcct 3120

gctcagctct gcataagtaa ttcaagaaat gggaggcttc accttaaaaa cagtgtgcaa 3180

atggcagcta gaggttttga taggaagtat gtttgtttct tagtgtttac aaatattaag 3240

tactcttgat acaaaatata cttttaaact tcataacctt tttataaaag ttgttgcagc 3300

aaaataatag cctcggttct atgcatatat ggattagcta taaaaaatgt caataagatt 3360

gtacaaggaa aattagagaa agtcacattt agggtttatt ttttacactt ggccagtaaa 3420

atagggtaaa tcctattaga attttttaaa gaactttttt taagtttcct aaatctgtgt 3480

gtgtattgtg aagtggtata agaaatgact ttgaaccact ttgcaattgt agattcccaa 3540

caataaaatt gaagataagc tc 3562

<210> 4

<211> 574

<212> PRT

<213> human (human)

<400> 4

Met Cys Pro Arg Ala Ala Arg Ala Pro Ala Thr Leu Leu Leu Ala Leu

1 5 10 15

Gly Ala Val Leu Trp Pro Ala Ala Gly Ala Trp Glu Leu Thr Ile Leu

20 25 30

His Thr Asn Asp Val His Ser Arg Leu Glu Gln Thr Ser Glu Asp Ser

35 40 45

Ser Lys Cys Val Asn Ala Ser Arg Cys Met Gly Gly Val Ala Arg Leu

50 55 60

Phe Thr Lys Val Gln Gln Ile Arg Arg Ala Glu Pro Asn Val Leu Leu

65 70 75 80

Leu Asp Ala Gly Asp Gln Tyr Gln Gly Thr Ile Trp Phe Thr Val Tyr

85 90 95

Lys Gly Ala Glu Val Ala His Phe Met Asn Ala Leu Arg Tyr Asp Ala

100 105 110

Met Ala Leu Gly Asn His Glu Phe Asp Asn Gly Val Glu Gly Leu Ile

115 120 125

Glu Pro Leu Leu Lys Glu Ala Lys Phe Pro Ile Leu Ser Ala Asn Ile

130 135 140

Lys Ala Lys Gly Pro Leu Ala Ser Gln Ile Ser Gly Leu Tyr Leu Pro

145 150 155 160

Tyr Lys Val Leu Pro Val Gly Asp Glu Val Val Gly Ile Val Gly Tyr

165 170 175

Thr Ser Lys Glu Thr Pro Phe Leu Ser Asn Pro Gly Thr Asn Leu Val

180 185 190

Phe Glu Asp Glu Ile Thr Ala Leu Gln Pro Glu Val Asp Lys Leu Lys

195 200 205

Thr Leu Asn Val Asn Lys Ile Ile Ala Leu Gly His Ser Gly Phe Glu

210 215 220

Met Asp Lys Leu Ile Ala Gln Lys Val Arg Gly Val Asp Val Val Val

225 230 235 240

Gly Gly His Ser Asn Thr Phe Leu Tyr Thr Gly Asn Pro Pro Ser Lys

245 250 255

Glu Val Pro Ala Gly Lys Tyr Pro Phe Ile Val Thr Ser Asp Asp Gly

260 265 270

Arg Lys Val Pro Val Val Gln Ala Tyr Ala Phe Gly Lys Tyr Leu Gly

275 280 285

Tyr Leu Lys Ile Glu Phe Asp Glu Arg Gly Asn Val Ile Ser Ser His

290 295 300

Gly Asn Pro Ile Leu Leu Asn Ser Ser Ile Pro Glu Asp Pro Ser Ile

305 310 315 320

Lys Ala Asp Ile Asn Lys Trp Arg Ile Lys Leu Asp Asn Tyr Ser Thr

325 330 335

Gln Glu Leu Gly Lys Thr Ile Val Tyr Leu Asp Gly Ser Ser Gln Ser

340 345 350

Cys Arg Phe Arg Glu Cys Asn Met Gly Asn Leu Ile Cys Asp Ala Met

355 360 365

Ile Asn Asn Asn Leu Arg His Thr Asp Glu Met Phe Trp Asn His Val

370 375 380

Ser Met Cys Ile Leu Asn Gly Gly Gly Ile Arg Ser Pro Ile Asp Glu

385 390 395 400

Arg Asn Asn Gly Thr Ile Thr Trp Glu Asn Leu Ala Ala Val Leu Pro

405 410 415

Phe Gly Gly Thr Phe Asp Leu Val Gln Leu Lys Gly Ser Thr Leu Lys

420 425 430

Lys Ala Phe Glu His Ser Val His Arg Tyr Gly Gln Ser Thr Gly Glu

435 440 445

Phe Leu Gln Val Gly Gly Ile His Val Val Tyr Asp Leu Ser Arg Lys

450 455 460

Pro Gly Asp Arg Val Val Lys Leu Asp Val Leu Cys Thr Lys Cys Arg

465 470 475 480

Val Pro Ser Tyr Asp Pro Leu Lys Met Asp Glu Val Tyr Lys Val Ile

485 490 495

Leu Pro Asn Phe Leu Ala Asn Gly Gly Asp Gly Phe Gln Met Ile Lys

500 505 510

Asp Glu Leu Leu Arg His Asp Ser Gly Asp Gln Asp Ile Asn Val Val

515 520 525

Ser Thr Tyr Ile Ser Lys Met Lys Val Ile Tyr Pro Ala Val Glu Gly

530 535 540

Arg Ile Lys Phe Ser Thr Gly Ser His Cys His Gly Ser Phe Ser Leu

545 550 555 560

Ile Phe Leu Ser Leu Trp Ala Val Ile Phe Val Leu Tyr Gln

565 570

<210> 5

<211> 1770

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 5

cagatgagga aagtgaagga gactgaggtt gcactgataa cctcagtgga tttaggtacc 60

gaggaagtaa aatgacctta gcagccaaac caagtgggcg actgcccaga gctggggtta 120

tgcaggaggc cgcacgagga agggacagac agggctttct gaagaaatct gctgtgactc 180

tgagggtggg aatatcatac agatggtact caacgccacc cctgctctac ctgaagtcct 240

gtgcacagag taaatccaga acctgctgag gtatcttgtt cacctcacca gcaagtcctt 300

ccttagtcta agctgctgtg gatggggctg tctgagagct tccttttaaa gtcttcgact 360

tggagaacga caaaagaatt ctaggacaag gcaggtgtac agtgtacact ttacaagtgt 420

aaagcagggc tgtagtagat tagtttatga ggaagaaagt tccattaaaa tgtaagcatg 480

ggagagaaca agtgcttagg aggagggtac acactcttgg gttcgggctg ctcaaaataa 540

gagatgtcct tgtcattgca acagccacac atactggttt tggtggcttc tgaagctgca 600

gctccaaagg tagctaagga aacttttccc ttttttggta aatgagatca tagggtggtt 660

cttatgtgca ctggctagga ttacagctgg cacggtaaaa ctcgagatca caggagctat 720

aggggaacat taagctatct ttactgtaaa caaagtttta gttttagttg ggagacttcc 780

agaaaatacc taggaaagga taaaagcaag ccttaagcac agttcattgc tattttaaca 840

tttctatttg ataacaaaaa ttttcctata taaaaggcct attgtccctc tgtctgcaac 900

cttcacagcc aactttgttc attgggctga aattcttgat tctcagctgg cagaaggctt 960

cctccagacg ccaaagcaag tgacttcttt ctctttccac gtacctgtaa tgttctcatg 1020

gctttctatc ctgctcgggg tacaggggtg atcaatgccc agtctaacag agacttctgt 1080

caggcctgat tcgcctagat gaattggaga agtccttgag tatagcagtt gtgaggactt 1140

gagatgtgac ccaagggaag caatgaaagg gggttctggg ggtgtctgca ttctggtctg 1200

ggcctgacct gtgggctaac tcccgaaaac aatgaagatg agctgggtag gccctggtaa 1260

aggtggtctg tgagccatct ggctggaggg ggctgggaca gggagtccca aatgcacaca 1320

ttcacagcag cttcactgga tgaggcgccc agggcaacaa tgctgtacct acctgtgatc 1380

tacgtgtctc tgggctcagc ccaaggggcc cctttggagg taaggagaac actagaacat 1440

gtatgtggtc aggtgtcata gaggccagcc cattttggtg ccactgggct aacatgatat 1500

actttaaaat atgcttttat tggaaaattc cagatactta attctttttt tttctttttt 1560

cttttctgtt ttacttaggc actgggaaat catgaatttg ataacggtgt ggaaggactg 1620

attgatcccc tcctcagaaa cgttaaattt ccaattctga gcgcaaacat taaggcacgg 1680

gggcctctag cacatcagat atctggactt tttctgccat ctaaagttct ctctgttggc 1740

ggtgaggttg tggggattgt tggatatact 1770

<210> 6

<211> 1133

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 6

cacgaatggt cgatggctgt gtctccctta gtctgctgtt tggagaaact ctttttacac 60

tgggctttgc agacttccct cttgaaacct ctttatgaga cttttgcatt tgggacactt 120

tcattttagt gccggttctt agaaactaac ctgagagcat ctaggtcaac tggaggactg 180

ccttgcaata gagtgctggg tgagacttca ggacgccatg tcggtggtcc cctcgtgacc 240

actgaagact tgctaagttt gggctccagg cttggaggcc ttgtttatgt ctcagcatag 300

gattcccagg gcctatgcta cccagagccc aaggcactca ttattagcac tgggtgagct 360

ccatgaggtt cgcgaccctt cctagggcag gccatgttta gatgaggcct ctatgatgca 420

atgcagcaat cagttctact ccaatggtgc cacaccctgc ctgcaagtaa gcagttgtca 480

tctaccatgc taaaattagt ttccaagggc cagcaagatg gctctggggt aatgaagggg 540

actttccacc tcacctgata cctggagttc aaaccctgga acctacatgg taggaggaga 600

gatctgaaat ccctaagtat aaataaataa aaaaaacaaa taaataaaat gtaaaaacct 660

ccttttgaaa ttggctttca aaacctacat gttgacattt taggtctgaa gacattttag 720

gtttccaggc aaagcaatga cactggcagc cgttgtaaag accatgcctt tgtgctcaga 780

catgtttttc atggttagtg tgaaggccta aatgtaaaag gagctggaaa caaaccatac 840

ctgaagtctc tttgtagtat gagattcagt ttgtgcaaat atatcatcta tggcccagtg 900

agatggttca acagaaaatg atgcttgctg catcgcctga tgtcctgagt ttgatcccca 960

ggacccacat ggtggacaaa gagaaccaat tcttgaaaat tgtcctctga cctccacgca 1020

tgtattgtag tatgtatgtg caaacaaaaa taaatgttac tgtgatggtt tgtacatgct 1080

tggcccaggg aatggcacca ttagaaggtg tgaccttgtt ggagtaggtg tgc 1133

<210> 7

<211> 4354

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 7

cctttagtcc taataataag gtggcagaaa cagttggatc tctgtgttca aggctagcct 60

agtctgcata gagagttcct ggccggctaa ggtgcatagt aagatcctat tacaaaaata 120

aatgaatgag taaataaaag taaactacta ccgtagttgt gctggttagt tcttgccaac 180

ttaacacaaa cgagaatcac ctgagaagag gttcctcacg tgagcaggct ggactgtgcg 240

cacacatgtg ggacattttt ctgactggtg attgatgtga gagagcccag cccactgtag 300

gcggtgtcac ccatgggcag gtgggcctgg gtgacataag acaaacaagg cagtaaacag 360

cactcctctg tgtcctctcc atcagatttt ggtctggggt ttctgccctg actttgctca 420

gtgatacact gtcaactgga agaagcagca aatcctttcc tctccatgtt gtgctgtggt 480

cattgtgttt attacaataa atcaaaaaag agaaaaagac aagaaaacaa agtaaagcag 540

gctatgcttt tgccaagaaa acagccctcc caaaatgcac ttcacccaaa tactgccctc 600

tcccttctgt gtggtaattg cgacaatgcc tccctccttg cgcatggtat ttcagattct 660

aagggatggg gtaatgtgtc atacttttcc ttcctcaacg ctagtgttac caggcacgca 720

tgtggtcgct tcctcaacgg ctgactagct accctgagag ggcagaaaga aggccacagg 780

agtagggaag gcttatgcca gtgcttctgg cttggttgac tcaaattgtc cttctaggaa 840

gaacaatgtc cccagaactg ccattttggg gaaagggagc ctttggatct gacacttgag 900

gccacaggcc cagatgacaa gtctcagaga tagtttttcg ggtctggaaa gctagtggaa 960

aatggacaca tgacaactgt cacacagacc aaaagaaccc cacagatgtt tgaagtttgg 1020

agctgaagat gagggtgact aaagctattt ggtcctctat caagagcatt ggtgtccatg 1080

aacaggaatg ccctccccaa gggatacaaa atcaagatag tcacagcctg tactcaagtc 1140

agggtgggac ggaacaagat attgaagaag caacaaacaa aaacttgaac gttcaccata 1200

ccctaaggat agaacagaac tgatgagttg gttagtcttc cgtggaaaac tgtggatcca 1260

tgtgttgctg ttgtgcacgt ctgttcccag cattctagaa gctaatgcaa gactcagttc 1320

gaggccatcc tgagctacac agtgagaccc tgtctcaaaa aaaccccaaa acaagaaaca 1380

tccaccaaat acaaaactct gtacttaaga ttattacagg gctggagaga tggctcagtg 1440

gttaagagca ctgactgctc ttccagaggt cctgagttca attcccagca accacatggt 1500

ggctcataat catctgtggt gagatctgat gccctcttct ggagtgtctg aagatagcta 1560

caatgtactc ttataaataa aataaaaaaa tcttttttta aaaaattatt atataacaaa 1620

gggtaattta ttttaccata ggaagaaaac ttgatgtttt aagactagaa gaaagaaaaa 1680

caaatgtctg agactgagag accctttaga cctggatcag ctcttagaga agttaactag 1740

ctaggaggag gggagactgg gtatagcatc agaggtgggg aaaggtgtga gttcagcaac 1800

catgaacttt ccaggggaga cagggtggga ctgtggatag ctgcgtgctt cctagagagt 1860

tcactcagcc ctttgagtga caggtgctca gtcagcgcca gtgtgccaaa tgaggaaaca 1920

aggtgaacag gacaaaaagg ccaacataac ttgagagcac tcagtttagc ccggagacag 1980

acaacaaatg accccacaat atagttctgg aatgttctgg tgttacatga ctggtggtca 2040

tgtcccacca aagttttcta ggtttttgtc ttcttattaa aacgttctaa taaaggaact 2100

cagggataat aaaggaacaa caaggaaata gcattcagat ttctccctaa aagttcactt 2160

acgggacagt ttgccctatt caacatgtgc cccaaactag ctcaggccag attgcctccc 2220

taccctcata cctagacata actggagtat gactgaatag aaattgtttg atggttgttc 2280

aggagagaag gaacttaact ccattgtctg gagagagtgg tatgtgcagc tcagggaagg 2340

tggggtccta ggttgtcaag tgcattgttt ggagggtggt gtgtgcacat agtatgtgca 2400

gtaccaggct gccaagtgca gtgttagaag aggtgtatgt gcattgcaca agccaggttc 2460

attactaaac tatttgttgt gcttacttac ctgcctcact gggaagagga aacgaggtgg 2520

aggagaatgt accatggccc ttgttagcat caaggaccta gagccagtgc tgccaggcct 2580

ctctcagatg aggaaagtga aggagactga ggttgcactg ataacctcag tggatttagg 2640

taccgaggaa gtaaaatgac cttagcagcc aaaccaagtg ggcgactgcc cagagctggg 2700

gttatgcagg aggccgcacg aggaagggac agacagggct ttctgaagaa atctgctgtg 2760

actctgaggg tgggaatatc atacagatgg tactcaacgc cacccctgct ctacctgaag 2820

tcctgtgcac agagtaaatc cagaacctgc tgaggtatct tgttcacctc accagcaagt 2880

ccttccttag tctaagctgc tgtggatggg gctgtctgag agcttccttt taaagtcttc 2940

gacttggaga acgacaaaag aattctagga caaggcaggt gtacagtgta cactttacaa 3000

gtgtaaagca gggctgtagt agattagttt atgaggaaga aagttccatt aaaatgtaag 3060

catgggagag aacaagtgct taggaggagg gtacacactc ttgggttcgg gctgctcaaa 3120

ataagagatg tccttgtcat tgcaacagcc acacatactg gttttggtgg cttctgaagc 3180

tgcagctcca aaggtagcta aggaaacttt tccctttttt ggtaaatgag atcatagggt 3240

ggttcttatg tgcactggct aggattacag ctggcacggt aaaactcgag atcacaggag 3300

ctatagggga acattaagct atctttactg taaacaaagt tttagtttta gttgggagac 3360

ttccagaaaa tacctaggaa aggataaaag caagccttaa gcacagttca ttgctatttt 3420

aacatttcta tttgataaca aaaattttcc tatataaaag gcctattgtc cctctgtctg 3480

caaccttcac agccaacttt gttcattggg ctgaaattct tgattctcag ctggcagaag 3540

gcttcctcca gacgccaaag caagtgactt ctttctcttt ccacgtacct gtaatgttct 3600

catggctttc tatcctgctc ggggtacagg ggtgatcaat gcccagtcta acagagactt 3660

ctgtcaggcc tgattcgcct agatgaattg gagaagtcct tgagtatagc agttgtgagg 3720

acttgagatg tgacccaagg gaagcaatga aagggggttc tgggggtgtc tgcattctgg 3780

tctgggcctg acctgtgggc taactcccga aaacaatgaa gatgagctgg gtaggccctg 3840

gtaaaggtgg tctgtgagcc atctggctgg agggggctgg gacagggagt cccaaatgca 3900

cacattcaca gcagcttcac tggatgaggc gcccagggca acaatgctgt acctacctgt 3960

gatctacgtg tctctgggct cagcccaagg ggcccctttg gaggtaagga gaacactaga 4020

acatgtatgt ggtcaggtgt catagaggcc agcccatttt ggtgccactg ggctaacatg 4080

atatacttta aaatatgctt ttattggaaa attccagata cttaattctt tttttttctt 4140

ttttcttttc tgttttactt aggcactggg aaatcatgaa tttgataacg gtgtggaagg 4200

actgattgat cccctcctca gaaacgttaa atttccaatt ctgagcgcaa acattaaggc 4260

acgggggcct ctagcacatc agatatctgg actttttctg ccatctaaag ttctctctgt 4320

tggcggtgag gttgtgggga ttgttggata tact 4354

<210> 8

<211> 4567

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 8

ccagagccca aggcactcat tattagcact gggtgagctc catgaggttc gcgacccttc 60

ctagggcagg ccatgtttag atgaggcctc tatgatgcaa tgcagcaatc agttctactc 120

caatggtgcc acaccctgcc tgcaagtaag cagttgtcat ctaccatgct aaaattagtt 180

tccaagggcc agcaagatgg ctctggggta atgaagggga ctttccacct cacctgatac 240

ctggagttca aaccctggaa cctacatggt aggaggagag atctgaaatc cctaagtata 300

aataaataaa aaaaacaaat aaataaaatg taaaaacctc cttttgaaat tggctttcaa 360

aacctacatg ttgacatttt aggtctgaag acattttagg tttccaggca aagcaatgac 420

actggcagcc gttgtaaaga ccatgccttt gtgctcagac atgtttttca tggttagtgt 480

gaaggcctaa atgtaaaagg agctggaaac aaaccatacc tgaagtctct ttgtagtatg 540

agattcagtt tgtgcaaata tatcatctat ggcccagtga gatggttcaa cagaaaatga 600

tgcttgctgc atcgcctgat gtcctgagtt tgatccccag gacccacatg gtggacaaag 660

agaaccaatt cttgaaaatt gtcctctgac ctccacgcat gtattgtagt atgtatgtgc 720

aaacaaaaat aaatgttact gtgatggttt gtacatgctt ggcccaggga atggcaccat 780

tagaaggtgt gaccttgttg gagtaggtgt gccattgtgg atgtgggttt aagaccctca 840

ccctagctac ctgaaagtca gttttctgct agcagccttc agaagaagat gtagaactct 900

cagctctgcc tgcatcatgc atgcctagat gctgccaagt tcccaccttg atgataatgg 960

cctgaacctc tgaacctctg aacctgtagc cagccctaat taaatgttgt cctttataag 1020

acttgccatg gtcatagtat ctgtgcacag cagtaaaacc ctaagacagt tacatatata 1080

tatatattgg ttatatatgt gtatgtatat atacatatat atatgtatat gtgtatgtat 1140

atatacatat atatatatgt atatgtatat gtatatgtat atgtatatgt atatgtatat 1200

gtatatgtat atgtatatgt atatgtatag tttgtatgta ccatctacag ccttaaccca 1260

atgcaatatt aggtgaatgc aaccaagaac attgatttaa atttttcatg accactcaag 1320

cttttgtaca aaatgtattc atacattttg ctgttagtaa gtgaagagag ctttaacaag 1380

agattgtatt agtgtgttgg tatataaata gtacaatgaa attcttttaa tatccatttc 1440

attaatattc tttagggaca aatttagtct ttgaagatga aatctctgca ttgcagcctg 1500

aagtagataa actaaagacg ctaaatgtga ataagatcat cgccctgggg cactctggtt 1560

ttgagatgga caaacttatc gctcagaaag ttcgaggtgt ggacatcgtg gtgggaggac 1620

actccaacac ctttctctac acaggtaatt gttttgatgc gttctgcaag ctaggatgtt 1680

cattcagtgt gcagtatatg atgtattagc tgtaatatgc aggattatgt gttatacttt 1740

gcattagatg aggacaggta atcctgctgg gagacgttcc aagaattcat tcatgtcatg 1800

attaaaagtc tacactacaa gcacatagca cacataatga aactaaaaaa attattgatc 1860

tccagacaaa ggtcagccaa cggtagaatt tttaaaagat gaaatttttt tttagatatt 1920

ttcagtggtt cttggacctg aatccagacc tttctgtatt ttaaacacct atattcccag 1980

cttttggttt tctaagacag agtttcacta tctagccctg attggcccca ttctcccacc 2040

tctcaggaga atggtaggac tataagcata cacctctgta tctggtttgg aaagtttcat 2100

catcatcatc attttcatag aacttcttag aatttttaaa tgctgaatat atattacatt 2160

tggaatcagg aagtgagata cagtacaaat tatagaaaga aaagaaacac gatattgagt 2220

caaagtcctc attcctaact cacttgaatg accttgaaca ggcaagtcac ctgaccctct 2280

gggcactttg gctttctaat aagaagtctg tcaccgtttt gaagattaag agtctgggaa 2340

cagttgactt cactggtttg gccttgggtg aatgtccccc tggctcttca tcatatcaca 2400

acatgggaga gaatcaggaa gggacatgta cagggagccc aggtgtgtgt ggtggccctc 2460

ctgttacaac cactccctca catagaacta actgtggtct caaaagaact actctcctct 2520

gagggtgcta gctccagtga ccttgtcacc tcacattaga cactccttga aggtttcacc 2580

gtccctcagt gtcaccacag tgaggactaa taagctgcta acacatgaac acacactaga 2640

caaaccacat ccaagacaca acagtttcag gtgagtcttc atacttgcct ggtcttatta 2700

caattctaca gcaatcctgt ggatatgacc tattcaaact ctgggggctt tgtagctctt 2760

ccccaaggga gttttagaag gctacaacac acagcagaaa ccagttgatg ctcctgccac 2820

ttcacccttt atgttggctt gagccacagt tagcatgaca ttttacaaga ggatctaaca 2880

ggtatacgtg tacagacttg tgcactctca gagatatgta taaagtaaac cgaaaatccc 2940

caaagcaccc agactatcct gcttgtattg ttgttgttct tcagtttctt ttcttgttat 3000

tttgtttctt tttaccttga gacaggatct tatattgttt gtttcatttt tatggagaca 3060

gagtctcact ttgctttgta gtcctggctg acaacgttat atgttgactt aaccttgact 3120

gtaaggtccc ccttctactg gtatgacagg aagcactaat gtcacaagaa taccatcaca 3180

cacacacaca cacacacaca cacacacaca gagccacccc ttactccaga tgatgcaatc 3240

acccccccct ttgtcacttt gatgtttctt taccatctgt gccactttgt ggcatcattt 3300

gggcacccac agtctgctgc accgtgtggc tactgattac ggcaccctgc tagcccaaga 3360

agctggaggg aatggtgatt atccttttgc tcttacaact attgccccag ccagcaagtc 3420

ctgcaacatg agagatctga gggtgagctg tagcttgctc atctcctgaa tccagtctct 3480

taaacgctga cccgagtcta agtcatttgt ttcacatttc ataatagatt accaagatta 3540

aacttgctcc catgaaaatg tgttttggct ctcttttgag gactaaatat agcaattaaa 3600

ggacatgata tgaaatgaga cctactcatg tatttaaaca aatttttgcc agccttaaaa 3660

aaaaaatttt ttttcagaca ggctgacaag aaaactcagc agataaaggt acttgctgta 3720

ctagcctaga gacctgagtc tgaacccata taagggcaga aggagagaaa ctaaagccca 3780

aaagttagtt gtcctctgac ctcatgtgta ccacagcata tcctcatatt tattctgtct 3840

ctctctctct cacttactca ctctctctct ctctctcact tactcactct ctctttctct 3900

ctcacataca cacacaaaca cacacacacc acacacactg aaaaaaactt aagcaatctt 3960

ttactaaacc attttatgaa attaccaaga agcaacaaga gagagagaga gagagagaga 4020

gagagagaga gagagagaga gagagagaga gaagaaagga aaggaaagga agaaaagaga 4080

agagaagaga agagaagaga agaaaagaaa agaaaagaaa agaaaagaaa agaaaagaaa 4140

agaaaagaaa agaaattacc aagaagctaa attaagtctg ttaaacatta catagtaaat 4200

gtagtaaaaa gaacacagct tgtggaagca tacatccttt tgtgttttaa gtccagatct 4260

tattgtctga tctggaattt actctgtagc ccaggctaga ctcaaatcat ggcagtcttt 4320

ctatgtaagc ctctgtagag ctgggaatag tggcatgagc catcatgctc agctagaaat 4380

ttaaagttta agaaatgtct cagcatgtga catgcacaca gcccaaagga aaacgtccct 4440

gaagcacctg aatagcacct gatagactct ggtcttttat ttagtctttt gtgtgtatta 4500

ctgcccacta aattagttcc cttcttgtct tgtttgttgt ttatttgtaa cccacattct 4560

aaaaacc 4567

<210> 9

<211> 1725

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 9

atgtgtcccc gagccgcgcg ggcgcccgcg acgctactcc tcgccctggg cgcggtgctg 60

tggcctgcgg ctggcgcctg ggagcttacg attttgcaca ccaacgacgt gcacagccgg 120

ctggagcaga ccagcgagga ctccagcaag tgcgtcaacg ccagccgctg catgggtggc 180

gtggctcggc tcttcaccaa ggttcagcag atccgccgcg ccgaacccaa cgtgctgctg 240

ctggacgccg gcgaccagta ccagggcact atctggttca ccgtgtacaa gggcgccgag 300

gtggcgcact tcatgaacgc cctgcgctac gatgccatgg cactgggaaa tcatgaattt 360

gataatggtg tggaaggact gatcgagcca ctcctcaaag aggccaaatt tccaattctg 420

agtgcaaaca ttaaagcaaa ggggccacta gcatctcaaa tatcaggact ttatttgcca 480

tataaagttc ttcctgttgg tgatgaagtt gtgggaatcg ttggatacac ttccaaagaa 540

accccttttc tctcaaatcc agggacaaat ttagtgtttg aagatgaaat cactgcatta 600

caacctgaag tagataagtt aaaaactcta aatgtgaaca aaattattgc actgggacat 660

tcgggttttg aaatggataa actcatcgct cagaaagtga ggggtgtgga cgtcgtggtg 720

ggaggacact ccaacacatt tctttacaca ggcaatccac cttccaaaga ggtgcctgct 780

gggaagtacc cattcatagt cacttctgat gatgggcgga aggttcctgt agtccaggcc 840

tatgcttttg gcaaatacct aggctatctg aagatcgagt ttgatgaaag aggaaacgtc 900

atctcttccc atggaaatcc cattcttcta aacagcagca ttcctgaaga tccaagcata 960

aaagcagaca ttaacaaatg gaggataaaa ttggataatt attctaccca ggaattaggg 1020

aaaacaattg tctatctgga tggctcctct caatcatgcc gctttagaga atgcaacatg 1080

ggcaacctga tttgtgatgc aatgattaac aacaacctga gacacacgga tgaaatgttc 1140

tggaaccacg tatccatgtg cattttaaat ggaggtggta tccggtcgcc cattgatgaa 1200

cgcaacaatg gcacaattac ctgggagaac ctggctgctg tattgccctt tggaggcaca 1260

tttgacctag tccagttaaa aggttccacc ctgaagaagg cctttgagca tagcgtgcac 1320

cgctacggcc agtccactgg agagttcctg caggtgggcg gaatccatgt ggtgtatgat 1380

ctttcccgaa aacctggaga cagagtagtc aaattagatg ttctttgcac caagtgtcga 1440

gtgcccagtt atgaccctct caaaatggac gaggtatata aggtgatcct cccaaacttc 1500

ctggccaatg gtggagatgg gttccagatg ataaaagatg aattattaag acatgactct 1560

ggtgaccaag atatcaacgt ggtttctaca tatatctcca aaatgaaagt aatttatcca 1620

gcagttgaag gtcggatcaa gttttccaca ggaagtcact gccatggaag cttttcttta 1680

atatttcttt cactttgggc agtgatcttt gttttatacc aatag 1725

<210> 10

<211> 57

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 10

gctactaact tcagcctgct gaagcaggct ggagacgtgg aggagaaccc tggacct 57

<210> 11

<211> 9

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 11

ggaagcgga 9

<210> 12

<211> 5025

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 12

ggaagcggag ctactaactt cagcctgctg aagcaggctg gagacgtgga ggagaaccct 60

ggacctatgt gtccccgagc cgcgcgggcg cccgcgacgc tactcctcgc cctgggcgcg 120

gtgctgtggc ctgcggctgg cgcctgggag cttacgattt tgcacaccaa cgacgtgcac 180

agccggctgg agcagaccag cgaggactcc agcaagtgcg tcaacgccag ccgctgcatg 240

ggtggcgtgg ctcggctctt caccaaggtt cagcagatcc gccgcgccga acccaacgtg 300

ctgctgctgg acgccggcga ccagtaccag ggcactatct ggttcaccgt gtacaagggc 360

gccgaggtgg cgcacttcat gaacgccctg cgctacgatg ccatggcact gggaaatcat 420

gaatttgata atggtgtgga aggactgatc gagccactcc tcaaagaggc caaatttcca 480

attctgagtg caaacattaa agcaaagggg ccactagcat ctcaaatatc aggactttat 540

ttgccatata aagttcttcc tgttggtgat gaagttgtgg gaatcgttgg atacacttcc 600

aaagaaaccc cttttctctc aaatccaggg acaaatttag tgtttgaaga tgaaatcact 660

gcattacaac ctgaagtaga taagttaaaa actctaaatg tgaacaaaat tattgcactg 720

ggacattcgg gttttgaaat ggataaactc atcgctcaga aagtgagggg tgtggacgtc 780

gtggtgggag gacactccaa cacatttctt tacacaggca atccaccttc caaagaggtg 840

cctgctggga agtacccatt catagtcact tctgatgatg ggcggaaggt tcctgtagtc 900

caggcctatg cttttggcaa atacctaggc tatctgaaga tcgagtttga tgaaagagga 960

aacgtcatct cttcccatgg aaatcccatt cttctaaaca gcagcattcc tgaagatcca 1020

agcataaaag cagacattaa caaatggagg ataaaattgg ataattattc tacccaggaa 1080

ttagggaaaa caattgtcta tctggatggc tcctctcaat catgccgctt tagagaatgc 1140

aacatgggca acctgatttg tgatgcaatg attaacaaca acctgagaca cacggatgaa 1200

atgttctgga accacgtatc catgtgcatt ttaaatggag gtggtatccg gtcgcccatt 1260

gatgaacgca acaatggcac aattacctgg gagaacctgg ctgctgtatt gccctttgga 1320

ggcacatttg acctagtcca gttaaaaggt tccaccctga agaaggcctt tgagcatagc 1380

gtgcaccgct acggccagtc cactggagag ttcctgcagg tgggcggaat ccatgtggtg 1440

tatgatcttt cccgaaaacc tggagacaga gtagtcaaat tagatgttct ttgcaccaag 1500

tgtcgagtgc ccagttatga ccctctcaaa atggacgagg tatataaggt gatcctccca 1560

aacttcctgg ccaatggtgg agatgggttc cagatgataa aagatgaatt attaagacat 1620

gactctggtg accaagatat caacgtggtt tctacatata tctccaaaat gaaagtaatt 1680

tatccagcag ttgaaggtcg gatcaagttt tccacaggaa gtcactgcca tggaagcttt 1740

tctttaatat ttctttcact ttgggcagtg atctttgttt tataccaata gggcgcgccc 1800

agggagtctc cttgtccttg atgtcaaact acatttttct tccagtgata ttcatatctg 1860

cctctggaga cctggctttg taacagcact catcatcctc aaggttccta gcagatgttc 1920

ttcacaagga agagactgta acatcatttg ttggggccag caactcagtg agcagataga 1980

aagtcacagt gaaccaacag ggtccttctg gcagggagtg ggtaagggga aacaactaga 2040

tgtagcttgc atagccacat aacacatctg gttaccattt ccctttctat tcatttctaa 2100

tccatcaaac aattgatgtt tacatacaac ttcatcatcg ccagctctgg tggcacatgc 2160

ctgtggtcac ggcacttggc agggaggaga ggatggctgc aagttctagg ccagcctgac 2220

atatgtagag tttcaggcca gtcagctaga tatcaagact cacacacaca aacaaacatt 2280

ataatttaca agtagatttt tgtagacaag tcttatgata agctaagcag aaagggtcga 2340

cttgttcaag gtcagacatc tccaaataat acatctaaga tctgtacctg agtcttttta 2400

cctcaagtcc aacctctcgc ctactgtcaa gtctcctctc ttcctgcggt tggtctcaga 2460

caacaaatct gtttctgcct tccatagtac cctttctttt gggctcttgt tgtctctcag 2520

gtttgagaga gtagctactg gacaggattc tttcctacaa caccatgtgc ctttggtgag 2580

tcatagacaa actgtataca gcagataata gattagtcca gggactgcaa aaggcagtca 2640

gggacaggca ggaaagggaa aggagaatat gaccaggact tacagaagag agtagatggg 2700

ttccataaac actagttaag gagcagaaca tggctccaaa gcacaacact ggttcttatt 2760

aactcctgtg cgactccaaa agccctgcct ctctccttcc ttcttcctca catggaggtg 2820

ccatgagaga acttcctaaa ataaactgta attctcaacc tgcacctgtc ctgtccaaga 2880

tgccaacacg gctctcattc aagtcacaca tgtacaatcc aatattctgg aaggcaagag 2940

tgaatttaag aagtggtata cataaatgtt tgaagtcact gagactcaaa tcccacacaa 3000

ccactgtaag gcatactcag gtcaagacat gagaagacca gcaggacagt gagccctatg 3060

ggaggcccaa gagataccca gactgatgaa atgatctgtg cccacagaac accagaccaa 3120

gacagtcctg tgaccaagtg agcatagtca gcactgtaca gtagttaaga atccttaact 3180

ataaaccagc agctagatgt ctggacaggg aacatgttgg tttcctagtg tttacgaata 3240

ttaaggactc ttgacccaaa ccgtttaaaa ttccaatcct tttatgaagt ttgttgcagt 3300

gaaacaatgc cttcagttct ttttgaatgt gtagattagt tataaacaga aatgtcagta 3360

agactgtaaa aaaaaatgaa gaaaggtgca tatgaggtat atttttatgc ttggcgagta 3420

aataaggaga aatcttatag cataattttt aaagaacatt tgtataattt ttctaaatat 3480

gtgtatatgt atattttatg cagcagtatt gagaaatgac cttggactat cttgtaattg 3540

taaattccaa aaataaagct gaagatatag tcctttgtct ttggtcttac ttcttctctt 3600

aacacttgta ccctcaggga tgcagcaaag acaatgttca cttcaaaact gagacgaaca 3660

atgctctaga attaagggtt ccggatcctc ggggacacca aatatggcga tctcggcctt 3720

ttcgtttctt ggagctggga catgtttgcc atcgatccat ctaccaccag aacggccgtt 3780

agatctgctg ccaccgttgt ttccaccgaa gaaaccaccg ttgccgtaac caccacgacg 3840

gttgttgcta aagaagctgc caccgccacg gccaccgttg tagccgccgt tgttgttatt 3900

gtagttgctc atgttatttc tggcacttct tggttttcct cttaagtgag gaggaacata 3960

accattctcg ttgttgtcgt tgatgcttaa attttgcact tgttcgctca gttcagccat 4020

aatatgaaat gcttttcttg ttgttcttac ggaataccac ttgccaccta tcaccacaac 4080

taactttttc ccgttcctcc atctctttta tatttttttt ctcgagggat ctttgtgaag 4140

gaaccttact tctgtggtgt gacataattg gacaaactac ctacagagat ttaaagctct 4200

aaggtaaata taaaattttt aagtgtataa tgtgttaaac tactgattct aattgtttgt 4260

gtattttaga ttccaaccta tggaactgat gaatgggagc agtggtggaa tgcctttaat 4320

gaggaaaacc tgttttgctc agaagaaatg ccatctagtg atgatgaggc tactgctgac 4380

tctcaacatt ctactcctcc aaaaaagaag agaaaggtag aagaccccaa ggactttcct 4440

tcagaattgc taagtttttt gagtcatgct gtgtttagta atagaactct tgcttgcttt 4500

gctatttaca ccacaaagga aaaagctgca ctgctataca agaaaattat ggaaaaatat 4560

tctgtaacct ttataagtag gcataacagt tataatcata acatactgtt ttttcttact 4620

ccacacaggc atagagtgtc tgctattaat aactatgctc aaaaattgtg tacctttagc 4680

tttttaattt gtaaaggggt taataaggaa tatttgatgt atagtgcctt gactagagat 4740

cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct 4800

ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc 4860

ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 4920

actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatctg 4980

acatggtaag taagcttggg ctgcaggtcg agggacctaa cgcgt 5025

<210> 13

<211> 7174

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 13

ggaagcggag ctactaactt cagcctgctg aagcaggctg gagacgtgga ggagaaccct 60

ggacctatgt gtccccgagc cgcgcgggcg cccgcgacgc tactcctcgc cctgggcgcg 120

gtgctgtggc ctgcggctgg cgcctgggag cttacgattt tgcacaccaa cgacgtgcac 180

agccggctgg agcagaccag cgaggactcc agcaagtgcg tcaacgccag ccgctgcatg 240

ggtggcgtgg ctcggctctt caccaaggtt cagcagatcc gccgcgccga acccaacgtg 300

ctgctgctgg acgccggcga ccagtaccag ggcactatct ggttcaccgt gtacaagggc 360

gccgaggtgg cgcacttcat gaacgccctg cgctacgatg ccatggcact gggaaatcat 420

gaatttgata atggtgtgga aggactgatc gagccactcc tcaaagaggc caaatttcca 480

attctgagtg caaacattaa agcaaagggg ccactagcat ctcaaatatc aggactttat 540

ttgccatata aagttcttcc tgttggtgat gaagttgtgg gaatcgttgg atacacttcc 600

aaagaaaccc cttttctctc aaatccaggg acaaatttag tgtttgaaga tgaaatcact 660

gcattacaac ctgaagtaga taagttaaaa actctaaatg tgaacaaaat tattgcactg 720

ggacattcgg gttttgaaat ggataaactc atcgctcaga aagtgagggg tgtggacgtc 780

gtggtgggag gacactccaa cacatttctt tacacaggca atccaccttc caaagaggtg 840

cctgctggga agtacccatt catagtcact tctgatgatg ggcggaaggt tcctgtagtc 900

caggcctatg cttttggcaa atacctaggc tatctgaaga tcgagtttga tgaaagagga 960

aacgtcatct cttcccatgg aaatcccatt cttctaaaca gcagcattcc tgaagatcca 1020

agcataaaag cagacattaa caaatggagg ataaaattgg ataattattc tacccaggaa 1080

ttagggaaaa caattgtcta tctggatggc tcctctcaat catgccgctt tagagaatgc 1140

aacatgggca acctgatttg tgatgcaatg attaacaaca acctgagaca cacggatgaa 1200

atgttctgga accacgtatc catgtgcatt ttaaatggag gtggtatccg gtcgcccatt 1260

gatgaacgca acaatggcac aattacctgg gagaacctgg ctgctgtatt gccctttgga 1320

ggcacatttg acctagtcca gttaaaaggt tccaccctga agaaggcctt tgagcatagc 1380

gtgcaccgct acggccagtc cactggagag ttcctgcagg tgggcggaat ccatgtggtg 1440

tatgatcttt cccgaaaacc tggagacaga gtagtcaaat tagatgttct ttgcaccaag 1500

tgtcgagtgc ccagttatga ccctctcaaa atggacgagg tatataaggt gatcctccca 1560

aacttcctgg ccaatggtgg agatgggttc cagatgataa aagatgaatt attaagacat 1620

gactctggtg accaagatat caacgtggtt tctacatata tctccaaaat gaaagtaatt 1680

tatccagcag ttgaaggtcg gatcaagttt tccacaggaa gtcactgcca tggaagcttt 1740

tctttaatat ttctttcact ttgggcagtg atctttgttt tataccaata gggcgcgccc 1800

agggagtctc cttgtccttg atgtcaaact acatttttct tccagtgata ttcatatctg 1860

cctctggaga cctggctttg taacagcact catcatcctc aaggttccta gcagatgttc 1920

ttcacaagga agagactgta acatcatttg ttggggccag caactcagtg agcagataga 1980

aagtcacagt gaaccaacag ggtccttctg gcagggagtg ggtaagggga aacaactaga 2040

tgtagcttgc atagccacat aacacatctg gttaccattt ccctttctat tcatttctaa 2100

tccatcaaac aattgatgtt tacatacaac ttcatcatcg ccagctctgg tggcacatgc 2160

ctgtggtcac ggcacttggc agggaggaga ggatggctgc aagttctagg ccagcctgac 2220

atatgtagag tttcaggcca gtcagctaga tatcaagact cacacacaca aacaaacatt 2280

ataatttaca agtagatttt tgtagacaag tcttatgata agctaagcag aaagggtcga 2340

cttgttcaag gtcagacatc tccaaataat acatctaaga tctgtacctg agtcttttta 2400

cctcaagtcc aacctctcgc ctactgtcaa gtctcctctc ttcctgcggt tggtctcaga 2460

caacaaatct gtttctgcct tccatagtac cctttctttt gggctcttgt tgtctctcag 2520

gtttgagaga gtagctactg gacaggattc tttcctacaa caccatgtgc ctttggtgag 2580

tcatagacaa actgtataca gcagataata gattagtcca gggactgcaa aaggcagtca 2640

gggacaggca ggaaagggaa aggagaatat gaccaggact tacagaagag agtagatggg 2700

ttccataaac actagttaag gagcagaaca tggctccaaa gcacaacact ggttcttatt 2760

aactcctgtg cgactccaaa agccctgcct ctctccttcc ttcttcctca catggaggtg 2820

ccatgagaga acttcctaaa ataaactgta attctcaacc tgcacctgtc ctgtccaaga 2880

tgccaacacg gctctcattc aagtcacaca tgtacaatcc aatattctgg aaggcaagag 2940

tgaatttaag aagtggtata cataaatgtt tgaagtcact gagactcaaa tcccacacaa 3000

ccactgtaag gcatactcag gtcaagacat gagaagacca gcaggacagt gagccctatg 3060

ggaggcccaa gagataccca gactgatgaa atgatctgtg cccacagaac accagaccaa 3120

gacagtcctg tgaccaagtg agcatagtca gcactgtaca gtagttaaga atccttaact 3180

ataaaccagc agctagatgt ctggacaggg aacatgttgg tttcctagtg tttacgaata 3240

ttaaggactc ttgacccaaa ccgtttaaaa ttccaatcct tttatgaagt ttgttgcagt 3300

gaaacaatgc cttcagttct ttttgaatgt gtagattagt tataaacaga aatgtcagta 3360

agactgtaaa aaaaaatgaa gaaaggtgca tatgaggtat atttttatgc ttggcgagta 3420

aataaggaga aatcttatag cataattttt aaagaacatt tgtataattt ttctaaatat 3480

gtgtatatgt atattttatg cagcagtatt gagaaatgac cttggactat cttgtaattg 3540

taaattccaa aaataaagct gaagatatag tcctttgtct ttggtcttac ttcttctctt 3600

aacacttgta ccctcaggga tgcagcaaag acaatgttca cttcaaaact gagacgaaca 3660

atgctctaga attaagggtt ccggatcctc ggggacacca aatatggcga tctcggcctt 3720

ttcgtttctt ggagctggga catgtttgcc atcgatccat ctaccaccag aacggccgtt 3780

agatctgctg ccaccgttgt ttccaccgaa gaaaccaccg ttgccgtaac caccacgacg 3840

gttgttgcta aagaagctgc caccgccacg gccaccgttg tagccgccgt tgttgttatt 3900

gtagttgctc atgttatttc tggcacttct tggttttcct cttaagtgag gaggaacata 3960

accattctcg ttgttgtcgt tgatgcttaa attttgcact tgttcgctca gttcagccat 4020

aatatgaaat gcttttcttg ttgttcttac ggaataccac ttgccaccta tcaccacaac 4080

taactttttc ccgttcctcc atctctttta tatttttttt ctcgagggat ctttgtgaag 4140

gaaccttact tctgtggtgt gacataattg gacaaactac ctacagagat ttaaagctct 4200

aaggtaaata taaaattttt aagtgtataa tgtgttaaac tactgattct aattgtttgt 4260

gtattttaga ttccaaccta tggaactgat gaatgggagc agtggtggaa tgcctttaat 4320

gaggaaaacc tgttttgctc agaagaaatg ccatctagtg atgatgaggc tactgctgac 4380

tctcaacatt ctactcctcc aaaaaagaag agaaaggtag aagaccccaa ggactttcct 4440

tcagaattgc taagtttttt gagtcatgct gtgtttagta atagaactct tgcttgcttt 4500

gctatttaca ccacaaagga aaaagctgca ctgctataca agaaaattat ggaaaaatat 4560

tctgtaacct ttataagtag gcataacagt tataatcata acatactgtt ttttcttact 4620

ccacacaggc atagagtgtc tgctattaat aactatgctc aaaaattgtg tacctttagc 4680

tttttaattt gtaaaggggt taataaggaa tatttgatgt atagtgcctt gactagagat 4740

cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct 4800

ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc 4860

ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 4920

actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatctg 4980

acatggtaag taagcttggg ctgcaggtcg agggacctaa cgcgtcacga atggtcgatg 5040

gctgtgtctc ccttagtctg ctgtttggag aaactctttt tacactgggc tttgcagact 5100

tccctcttga aacctcttta tgagactttt gcatttggga cactttcatt ttagtgccgg 5160

ttcttagaaa ctaacctgag agcatctagg tcaactggag gactgccttg caatagagtg 5220

ctgggtgaga cttcaggacg ccatgtcggt ggtcccctcg tgaccactga agacttgcta 5280

agtttgggct ccaggcttgg aggccttgtt tatgtctcag cataggattc ccagggccta 5340

tgctacgaag ttcctattct ctagaaagta taggaacttc aggtctgaag aggagtttac 5400

gtccagccaa gctagcttgg ctgcaggtcg tcgaaattct accgggtagg ggaggcgctt 5460

ttcccaaggc agtctggagc atgcgcttta gcagccccgc tgggcacttg gcgctacaca 5520

agtggcctct ggcctcgcac acattccaca tccaccggta ggcgccaacc ggctccgttc 5580

tttggtggcc ccttcgcgcc accttctact cctcccctag tcaggaagtt cccccccgcc 5640

ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac tagtctcgtg 5700

cagatggaca gcaccgctga gcaatggaag cgggtaggcc tttggggcag cggccaatag 5760

cagctttgct ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc cgggggcggg 5820

ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt cctccggagg cccggcattc 5880

tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc tccgggcctt 5940

tcgacctgca gcctgttgac aattaatcat cggcatagta tatcggcata gtataatacg 6000

acaaggtgag gaactaaacc atgggatcgg ccattgaaca agatggattg cacgcaggtt 6060

ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct 6120

gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga 6180

ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg 6240

ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact 6300

ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg 6360

agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct 6420

gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg 6480

gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt 6540

tcgccaggct caaggcgcgc atgcccgacg gcgatgatct cgtcgtgacc catggcgatg 6600

cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc 6660

ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag 6720

agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt 6780

cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgaggggat caattctcta 6840

gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct 6900

cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg 6960

aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc 7020

aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct 7080

ctatggcttc tgaggcggaa agaaccagct ggggctcgac tagagcttgc ggaacccttc 7140

gaagttccta ttctctagaa agtataggaa cttc 7174

<210> 14

<211> 25

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 14

gcagtaaatg agagctgcgt gaggg 25

<210> 15

<211> 3732

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 15

ggcagtgttg gggtaggtta tggagagaga agaaaaatgt ctggaagccc gggagcctgg 60

catggctgtc ctctgagagg ccctactgat tgagcatctg actgagacag aatcagatgc 120

ttacacccaa ccattgggct gaagttgggg atccctatga ttgaattagg ggaaggattg 180

aagaagctga aggggagggc aaccctatag gaagaccaac agtttcaact aaaccagacc 240

agaccccaag gagctcccag agactgagcc accaacaagg cagcatacac cagcaggtca 300

aagatccacc ctggcacata tgtagcagag gtctacctag tctggcctca gtgggagaag 360

atttgcttga tccttgagag acttgagacc ccaggaaagg aggaggccgg gtgcagtgca 420

ggggaagcac tctttcagag tcaaggagga agagggggaa tgggatgagg aactgtggga 480

ggggggacag agggagggca atgcctggaa tgtaaataag taaaataatt aaaaaaaaga 540

aaaggaaaag aaaaaaaaat gtctggaagc catatatgag atgtaaagac tattgaaaat 600

ttatgagttg tggctgaaca aagcaggtgg acagcatcct accgcaaggg gccagataga 660

aagccacccc agagttccag cagactctgt gggatacaaa ggtccagtgg taaccactgg 720

accaaggcac acactctcag tccggtgaca gtgagccagc cttatcttca tagcgttttg 780

ccccaccctg cttactttag gaacaagagt tttataagtt ggactactca ggcagtgaca 840

tcactagctc agcatgccaa gggatcactg acttagtata catgcaggaa gtaggttctt 900

ttctagataa cactagtttg ccccaaaccc agtgtcttga aagggagtca gttgacctcc 960

agatagccct tatcaaaatg ttcttccttc attgtttcct tgagtgggta ggagaacggg 1020

gaatgttgat gctccacttc cctgtgtgat cacacaccac tgaacattca ataatatctt 1080

caaattatgt atttgcatct tgtgaattcc tacagacttg aggatgtagc tcggttggta 1140

gagtgattgt ctggcataca ggaagcccag ggttcaatcc ccactactgt gtgaactgca 1200

cagtgacaca tgcttttaac cccaacacta gaaagacaga cacaagagga aagagctcca 1260

ggccatagat gacagtacaa tgagctggag ggcatcttga actgcacgta accttgtcaa 1320

agaagaagaa aaagatttct acagttcaat taaagcagat gcaaatgaca gggacagaag 1380

gacaacaaaa tggcttcatg ggtcaaggta cttacaagcc tcgtgacctg agtgtcatcg 1440

ctagaactta cattgtgaga aaaatctgcc ctgaaaattg tcttctgatc tttacacatg 1500

tgttatatgt gcacactgca cacatacaca cgagcgaaat taataaataa atgcattttt 1560

ttcaattggg gctttccccc cctccctttt ttgagacagg gtttctctgt gtagccctgg 1620

ctatcctgtt actcattctg tagaccaggc taccctcaaa ctcagagatt tgcttgcctt 1680

tgcctcacaa gcactgggat taaaggtatg tgccatcacc accaaactaa taaaaaaaaa 1740

tttaaaggaa atgtctacaa gaaaggacac aaatggtcca taagtttcta tttgtgtttc 1800

ccagttggtt tgacatgctg accatctttc aaaccaactg ggaaacacac atagaaacta 1860

cagtgagata tcacatgtgc cataaggtga ctatcattta agacaaaaaa aaaatagagt 1920

tggtgaagat gtacagagac ttttcgttgc atgtgagaac aagcaacata gcaggtcctt 1980

acttcacctc ccaactaaac tatttcatca gccgtgaacc ctggtctgat tctggtttca 2040

taccttatga ataagctgct atgaatgtga gcattcaaat acacacctag tgtgaagtgt 2100

tggcttttga aagaaaggaa atgctgacac actgagtctt gaaggcattg caccaagtaa 2160

aacaaattcc gtcagaaaag aacaactatg aggtccttag aatgatcaaa tctatacaaa 2220

ctggtaatgc aatgttgact gccagaggct ggaaaggaag gaaatggaga ggtggttttg 2280

gttagttggt agtttagttt gtttgtgttg ttgttgttgc tgctgctgtt gttgaggcag 2340

gcttcactaa ctcactacat gaccagactg ttttcctgcc gggactccca agaactgtaa 2400

ttataggtga ttgccaccgt gttgtgtggg agtcactgtt taatgagttt acaatgcaat 2460

cttggaaaga taaaaatgtt ctgggaaatg aatgatggca aatatctgta aacttaggac 2520

agtacgcggt aatgcacatc ctgtatatat gatgtatatt ttattgcaat tgaaaagcaa 2580

cttggttgtg ggtgagatag ggcctcacta tgtaccccat actttcaact ctcaactcag 2640

tctcctgagt gctaggatta caggtatgcc ccaatgtagt gtgtgtgtgt gtgtgtgtgt 2700

gtgtgtgtgt gtgtgtgtaa caatgcagac ctggtgcatt ctggaattgt gtgggcatta 2760

gataacttct ataatgacca gaacacacgg gaacctcacc aatactcttt cctttctcag 2820

aactactagc gtgttgacta tacttcattc aaacacaggg gttaatccat tagacaggga 2880

atttgtgctg tggcgattta tatttaatga tttgttcata acaaatatgg atccccacta 2940

ttactgctaa aagctttaca caaacacaca cacacacaca cacacacaca aacacacaca 3000

cacacaaaca cacacacacc acaaaaggta agggaggagc gccccagagt agctgtcagg 3060

tttacccgct aggatccgcc ttagctttca gagctggccc tatgccaccg ctgagggtcc 3120

caaagattgc tttgctcctg agattccagc tcgaatacgt ggagcggttg cgtcctcaaa 3180

cctaagtagc gtctcggccg gtgtgtcccg gtcccaccga agcaaccttc cagcgcacct 3240

ctcaggcggg ctctggaccc ctctcactcc cacgtttgtc ttcctttcct tccccccccc 3300

accccccccg ccccagcccc ttctttttcg gcgaccgagc caccgagcaa cacaagccgc 3360

tgtacactct tctggagaaa tcggtgtaat agagagtgag gatccggaac gctgcccctg 3420

cagttgtcac cgccgcccgc ccggggactg cggcacccag cctctcggac ctgctcgctc 3480

accaggcgaa caacctcctc tctgttccct gcgggctgcg tgtgctgggc cagaggctcc 3540

cgggtgcctc gggtgccagg gtcgcatcgg gtgactcggg gagtgtgtct gcaagcggga 3600

gggcctgggg ctggaagagg cgggtctgtc cccgccccgc cctcctctac tccgcagttt 3660

agtagaggcc ccggtggccg gtgctcactg ggctcagggc cagtccaccc gctcaacgcg 3720

ctctagccag cc 3732

<210> 16

<211> 4951

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 16

aagtggcatc caaattttct tgctcaacct aaaaccacac ttaacatttt tattttagct 60

aacaaaatgt ggattattta tggtgtataa catggtgttt taagatgtgt accaacactg 120

tggaatggct gtgtcaagct aattagcaat gccttctctg tgcttgctct gaatttctca 180

gtggctatat ttctacaaga gtccttttcc ctgttcctcc tttgtatttc ctctctggta 240

ttccctttgt cttcttgata tacctctcta ttgtgattta gaaaactaaa cgtgtgatac 300

tgggctgaag agccagctca atggttaaga gcacctgacg cttttgcaga ggacccaggt 360

tcaaatccca gctcacaacc atatgggatt tagggccatg ttcttgcctt tgagcaccat 420

gcatgcaagt ggtacacaga caaatatgca gtcaaaacac acacgagcat aaaaaaatta 480

ccattatgat agtaagctga ggttggctta cacaatcacc tatgtatagc agcagtcagc 540

aggagacata cactgtggtg taccaacact tctttcactc tggcagttaa ctatacaaaa 600

actgagagat caccacaggc cacattcctg cctccaaatt cctgtccact ctgtctcagc 660

agcattttcc cataacacat cttctgtctg ccacattctt tgtgcctggt aatgctgcga 720

tcggacagga agctcaggga tgagtaagac atagctctgt gcccacagct tagtaaagag 780

agaaatccaa gcttctctct gtctaggagg cattcacact cttactttgg taggttctat 840

gcctcttctg gtccatctac tttaatacaa aagctatttt acaagactct agccacagaa 900

ggaaatatct aacctactga ctatagtgcc actcatcttg attgaagtta ttgatccatg 960

gataagaatt gttacccaaa ccaagacagt aagaagcctg cagtgacctt tctaatgtat 1020

atgtaaatgt aagacaagct tttccctgcc tcctgtgcag tgattccata gttgtcttat 1080

ctgtctcccc tacataaagg cacttgaaaa acagtagtca acaggtctga agctgagaca 1140

aaaatgacgt ggaaacccca tatgattcaa gagctgtaaa gctcatacta aagtcagctc 1200

tattcccacc cctctgtggc tggattacat aaggatgcac tttgtattct ctcaaaacaa 1260

gactgccagc tctgtcttct gcaaccgaga acccaaacca gctccaatta caagagaaca 1320

tccaaaagcg tgcctgaaca cacacatagg tgggcttaag aggtaggagt caaacctcaa 1380

cattctagat gttaagtaaa acctctcaaa cccctcaggg aaaagtacac agtttccttc 1440

tcatactact aaaatgtagc agaaaagcta gaaataacta aataggtcac acagtgcttg 1500

gagcttggag ctcattccct tgcagattta atttcaatta agtctctcat tttaccagac 1560

ttctgtgata ccctttacat cttgggctgg agagttcaca cagctgttaa aagtgatggc 1620

tgctcttgca gaggaccagg gttcaatttc cactcatgtg gtacctcaca agcatctgta 1680

actccagccc caggagatcc aatgccctct ccaacctcct caggcaccag acatacacat 1740

agtgtacata cacataaaat aaatcttaaa aatcatacta attaaatctt ttcaagtatg 1800

gatactagca ataatatatg taaataaata tatatgtggg tacagtgtaa tgtgtagaaa 1860

agagaacagg aaaggctaag tattagaggc caagaatata gattgaatat aggcaaaaga 1920

aatgtgttat ggaagtggat ataaagctaa atgtttttct aagtctaact ttgtcaaaag 1980

aatttcattt ctggtgctag ataagtacat ggaaatatat ttgatactga aagtataaga 2040

tccgatgtaa agcaccatgg ctgccgtccc agtatctatc agtgtgtgtc tttttttttt 2100

tacaacattt tttataataa agtgtattta aagaactttt aaaatgaact cttttgaaat 2160

attatggcac atgcatatta attccagtac tcatgagttc aaggacagcc ttgggctatg 2220

cagcaagttc tagtctatca taaaactctg tctcaaaacc caaaacatga accatgcaaa 2280

taaagtacct tggcaacatc cttttcaatg gtgagctctt gggccttgcc tccatgtcca 2340

caacgatgca catttctgca tgctgcactg tgccatggca cgcatgttga ggtcagagga 2400

caagccggag gggtctgctc tcttcttccg ccatgtggat tccagggatc aaactcaggt 2460

ccctagctta gcagcaaagt gattttcctt acagagaaat cttctcaggc tctcctttac 2520

ttttcttacg gtcatcaatt ctgttgcagt atggtcccac ccaaatgact cttttaatct 2580

aagtttcctc ctgaaaggct ctgcctccaa atatagtcac tgtgagattt atagttttaa 2640

cataggaatt tggaaaagac ataattcaga ccgtaacaat gagtgaggcc tacttataga 2700

aaatgtgata tataatatat atataaatgt tatatgtgta catatgtata acatatatac 2760

acattagtca tacaaagtta aataagtcag tcatattata aaaggaaaaa gaataattcc 2820

tatatctgca agatgttttg tatatagcat cacattgcat ttccatggta actgtgtggc 2880

cttgtcttgt aggcgatgat gttaggccag aaaactatca aggtggaagt ggatattagc 2940

ccagaaactt ggaataccta aaatacaatt tgcaaaacac ataaaattca agaagaatga 3000

agaccaaagt gtggacactt tgtccctcct tagaatgggg aacaaaatac ccatggaagg 3060

agttacagag acaaagtttg gagctgagac gaaaggatgg accatctaga gactgctaca 3120

tctggggatc catcccataa tcagcctcca aatgctgaca ccattgcata cgccagcaag 3180

attttgctga aaggacccag atatagctgt ctcttgtgag gctatgctgg tgcctggcaa 3240

acacagaagt ggatgctcac agtcagctat tggatggaac acagggcccc caatggagga 3300

gctagagaaa gtacccaaga aactaaaggg gtctgcaacc ctataggtgg aacaacaata 3360

tgaactaacc tgtacccccg gatctcgtgt ctctagctgc atatgaatca gaagatggcc 3420

tagtcggtca tcattgggaa gaggcgcccc ttggtcttgc aaactttata tgcctcagta 3480

caggggaata ccagggccaa gaagtgggag tgagtgggta ggggagttgg gggagggagg 3540

atatggggga cttttgggat agcatttgaa atgtaaatga agaaaatacc taattaaaaa 3600

aaaaaagaag aagaagaaaa tgtgttgtgt atgtatatac atgaaccagc atggctagac 3660

acacagaaga gagactttga accttggccc acttgcttct tcacactaaa tacaattgca 3720

tttgacgcaa aagaatacaa agactgtggg tcttacttgt ttgctttcat tcaaaaacag 3780

caacacaggg ctgcagaatg gttcagtaga caatgtgctt gccatgcaag aatgatgacc 3840

aaagtttgaa tttccagtgt ctacataaaa gtcagaaggt ttgtgggcat gcctataatc 3900

ccagtactcc ggaggcagag actggagagg gcatgagcag caaactaaaa gctccagagg 3960

cagcgagaga gcctgcctca gtaaacaaag tagacagtca tagagaaaga cacccagtac 4020

caacctcagg tcgccagaca aacgtgcaca cgtgggcaag agcatccaca cacatgtact 4080

caaacacaca caaacacatg cacactgcac atacagagag ggagggagag agaaggagag 4140

agagggagag agactcaaaa agccctagca ataagtggca tatgttttag taaaatcaag 4200

acagtaaata caaaaggccg taatacctaa accacttttt attgctattt gttagtcttt 4260

attaaaacaa aaaatatatt ccaaacacta aagtacatga tatttataaa aatatacaaa 4320

gtgaaaagta catttctatt attctggtat gttattctat accaagtgtt tacatccagg 4380

atgtcatgga ggtagcttcc ttttgtacct aaagtaaaaa taaatccatt taataggaga 4440

caatagcaca aggttcattt ttaattccta aaagctctac tatagtaata ctctatgtga 4500

atgtgcgtgt gtgcacacac acacacacac acacacacaa acacacacac acacagtaat 4560

ccacaccctt cctggcacaa acagcaatta atagcaatgt tttgtctctg acctaataaa 4620

agcctcagaa ctaaaagctt tctggtattc attctaaagc aatttaacat aaatattaac 4680

atttccaccc tcctttaaaa atacttttaa tactactaaa atttgaagtt tccaatcaat 4740

agatgaacag ccaaaatctg gtttccctac tctatagaag tgagttttca ttacaatgcc 4800

acaaccttga acagttcctg gttttgtttt tcatctaact aaatggatta gtcaaaaagt 4860

aaaggagcta gagagatggt tcagcagtta acagctcctg ctgtgcaatt atgagaacaa 4920

gaaatctgat tccaccaccc atgtgacaag c 4951

<210> 17

<211> 45928

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 17

atgtgtcccc gagccgcgcg ggcgcccgcg acgctactcc tcgccctggg cgcggtgctg 60

tggcctgcgg ctggcgcctg ggagcttacg attttgcaca ccaacgacgt gcacagccgg 120

ctggagcaga ccagcgagga ctccagcaag tgcgtcaacg ccagccgctg catgggtggc 180

gtggctcggc tcttcaccaa ggttcagcag atccgccgcg ccgaacccaa cgtgctgctg 240

ctggacgccg gcgaccagta ccagggcact atctggttca ccgtgtacaa gggcgccgag 300

gtggcgcact tcatgaacgc cctgcgctac gatgccatgg taagacccga gcccgcgccc 360

gggatagtag tcccggactg agagaggagc cgggctggaa aagcagcgga tggcagagtg 420

tggcaagcct aggtccaggg cgcggagaga tgtggggata aagtgagact ccggccagtg 480

tgccagctgg atgcatagaa gtcccttgga cgattcgtct taaacggacg ttattgcgcc 540

cccactatga gatgggagta tttagaggag caggacccag tcctgcctgc gaggggctta 600

aggtggggtg cgtagaagca ctgacagtta tccagagcta gggactgaat cggatccctc 660

acgcagctct catttactgc tagatctttc aaagaatgct ttatctcaat ctagatcttt 720

cgaaaaatgc tttatctcaa tcttattgaa agggaaaccg cgtcgaagaa gctgaataaa 780

ttaacaggca ccatccccgc aaaaagggag acgtgataag aatggacacc agatctcttt 840

gaccccggag cttgagagta attaagctca cagctttggg gtagattctc tgttcaatta 900

ccttctatgt gaccttgagc aagttattta ctctttcttg cctcagttct atcctgtata 960

aaaataagga ttataattgt acatagtgca catagtgtgg tctctatatt attaccagac 1020

acatagtgtg gtgtctatat atgttgttat taccagacac tataatatag ggttaagaaa 1080

aggaaagaaa gaaagaaaga gaaagaaaga aagaaacttc agagagagcc aaaatgtgga 1140

aagacttggg tttcaaagga tttcaataaa gtaggtttgg cggcagagat ttaggaagag 1200

tccaacttaa gagataatgg gctcttaaaa catgtgcaaa agtttagttc aaagtcccaa 1260

atgttaggtt aaaggtaatg ccttttctat tgcactaaat tctagaaagg gagcagaggt 1320

ggttgcacag taataaataa gtacaaccaa gatgtatata tggtggttaa aaaaaaaata 1380

aaaccaccac ccaaatttaa ccctgtgtaa aagccttgag taaagggatt cagcaagact 1440

gtggagaaac ttgggagaag aaaaaagagc tttggaagag gaagaactga gaaatctcca 1500

gagaaacgtc cagagaaatc tgtgaagtcc acaagtgtgg aagggaaagg gaagtgtgat 1560

taggctggga tacagaaggc taacagtaca ctttatatga gaagagctta gaggtcagga 1620

gagcacaatg ctgaggggtt tgtttggttt tattttcccc tctggaaact tggagaaagg 1680

aggacaggaa aagatgataa gagggcacca ttgtttggtt ttaaaggtcc ctccccaacc 1740

ccccacaaca agaaagctca ttttaatgag atgagagaag gggagaaagc tccctccgtt 1800

ctgatgcttt tgagcttatc agggacaccc tgcagcagaa attccagctg atccttcagc 1860

cttcagagtt gatgaggtgg ggctcacaca catctggatt tgaagaagaa tcgtgggttt 1920

caagaactct gtggtctctt aagcattggt attccacagc tacacttccc tgatactttg 1980

cactcagtaa tgacaaaata aatatttgta gaattgagtt gttgctaccc tgtgtctcca 2040

cttcctccca gaagggtaga ataaacttac atgtgtttgt tttattttgc atataaacca 2100

cgactttgta aactttactt tttgtgacac tcttttctcc acaattgtgt gaaccctttt 2160

tagtggaaaa ataagaatga agaattccta gttttcccta ataatatttc cctcttgtaa 2220

gtctagttga gctcctggtg ttaatcatgg tatcaaatgt ttatgacagg acatctaggt 2280

gaggcctgag taaactgctg taagttacag cctatctctt ctgtgtttgt tcagaatcct 2340

ttttgttgga tgaaattact ttctttcaaa ttttgctcaa gaaaattgaa ttgggttctg 2400

aggaaaaata ggctacttac gtaatgttgg cagggggtaa agattgggag cggatcacat 2460

ggcaaggtta gatggctgtg tcctcaggac ttactctgtg cagtttgggg gtgtgtcaaa 2520

catgatttga cgtgcataaa atcccagttc tgtagtgcta aagcatgtgc taaagctctg 2580

ctgtactcca gtattgcaca gatgtttcac tgtcaattgt ttatattata ctaaccctca 2640

gagaatttat tgctgttgtg gtataaacgt tcctccactg taaccactca ccgcatattg 2700

attcctagga cactggggat tcaaatcagg gtggaaaaca gttgtggagg ggcatctcca 2760

gtttcctgga gactggacct agttcaaagg agccacataa aaggactgaa gcatatggag 2820

gacaaagagg aatttcctag aatccagcta aaacacagga gtcccctcga gcaggaactt 2880

ccaaggaggt gtacactttc tagacctact ggagtaatcc tgattgctgt agaggaccca 2940

gggcaatcct ggagaccccc tggaaccact actaggcaga aaactgggaa gaggatggca 3000

aaactctcca gttagtgtag ctgtctcagt ggaaggactg ttgtcccctg aggcacgagg 3060

aagacagcat ggaagcactg acagtatgcg cttggcctgg ttactttctc tgagcttcat 3120

gttccttagc tgtaaaacag gagtgataac ccccatcttg cagaggtttt gtgaggaata 3180

catgagataa tccctgctgg tgctcactga aagtgagttt catgtgccct gggtggggag 3240

gaggggaatt gtgaggaagg gaaggtttgg gttgtgcatc agattatctg attctcacag 3300

cctggcacag tctaccacct gcctgccctc taccatctta cctaagtacc attgcctaaa 3360

ctatttttgc tatagttact cattgagcag acttatgaaa tctccccaca gcacaaaact 3420

gaaggtatcc acctccagag gtgagattct caccaagaca ggctgtgggg aaggtagctt 3480

tgagcaggtc tctagggtcc cctagaaagg tgccttttct ctgcttggct gctgcttctg 3540

aggccctggc tgctgagccc acttcctgcc cataaggctg cagccttagc cctagccaat 3600

tgctcctaaa tgggggaggc agcagccagg gtgggaacca tggcccccgc ctttaactca 3660

cccaggggac aggttagtca gatagatggt cctttgttcg ccaccctcca cctcccacat 3720

agacaacatt ctaatgtgtt tgatggactg catatagttt tgtttctaca tgcactttca 3780

aaacctttgt gtgcttatat tttataatgg catacccagt cctgttattt agccctttct 3840

gtttcttact tttgtcctga acttattgtt tttatgattt gtccatgttg ccctagatgc 3900

ccttatacct atacatcact gtgaattgtg ctctgtgtat tttacccatc tccacgtatt 3960

ttacccatct gctccctggc tgatgggttt ggagatggcg tctaacaccc tgcctccaca 4020

aataatgctg ctccagatat cttcctggat ggtctcacca gatgatttct acccaggagc 4080

agaattgctg gccacagggc atactttgag tactacaaga ctggtttcct caatgactga 4140

catcctccat tctcttccac ctactcccat ccccaaacag gacatgtaaa tttctatatt 4200

ttacctatat atcatggcag tatccagctt tattgtaggt gtaaaataca cattgctgct 4260

ttatcatgta cttttctaat aactagtgag tttggacatt ttttgatatg tttgttaact 4320

ttttaagctt cctcttctct aattgcttat tcatattctt tggtattttt taaattgggt 4380

tagctttata ttttttgact ttcaggagtt ataggttttg atctgtgatc aattttagac 4440

attgcaaata tattatgatg atattaggtc tattaatttt gtccatggta tctttcattg 4500

aacagaaatt cttcatttct tgtaattgaa ttaatctttt tttccttgtg gcttgtgctt 4560

ttgaaagttt gcctaatagt ttgctaattt aaagcttatc acctttaaca ctcctcactg 4620

gaaattcaga gatcaccacg gttctctctg ccttttaaga gataaatcta caatctgtgg 4680

ctatgcaatt cacttctcag catcaccagt tcctcatcta taaaatagaa gtttgaacca 4740

gcttacctct gaggtcccat acatatttga acaactggag ccctacagtg aaatgttttg 4800

attcagtgaa gggacaggag gatggggcaa aggtgtagag gcaggaatct acacaatgga 4860

ccaggttgga ccagagtgag gtagcaggaa tcatggcaga agaggttgtg taaactctaa 4920

aggccacctg agaagtttga gttatgttgc catgggcagc catgtggttt ctgcgtagaa 4980

gaccactgtg gtataataaa aaataaatat ttggtccttg tccccagtac ctgtcacaga 5040

gttcctaaaa ccctagaaat ttcctaagta atagaaatgt cctttgtgat tcataataaa 5100

ccccatgata atacctgggt ttatgctcct gcagacttag ggtgagatcc ctagatagct 5160

tcaagatgga accagtcacc agcaagtcca agagattagg ggatcaaaga gttggaactt 5220

tcggtaccac ccaccaaagt ccaggaaagg agggtgggac tggagattca gctctacaaa 5280

aactcttgaa caagatgtga ggagcttttt agttgctgag catttggagg ttcctggagg 5340

gtgcacaccc aggtagggca tgaaagctcc gcattgcccc catatcttgg cctatccatt 5400

cttcacttgg ctgttcatct atatcctttg tatagccttt ataataaacc aatgagtgag 5460

ttctgtaagc tgtcctagca agttaattga agccaaggag gaagtaatgg aaacccctga 5520

tttatagcca ttcagccaga agcacaacct caggcttgtg atcagcctga agtcagggca 5580

gtcttgtagg attgagccct taacctatgg gatctgacaa tgtcttgaga tagtatcaga 5640

attgaattca attataggac actcagtttg tgttttctgg agtactgttt ggtgtctggg 5700

ggaaatccca cacacatctg gtcacagaag tattctgtgt tcagtgtgac agtagaaaaa 5760

gcagtttgtg tgtatgtgtg tgtttgtgct tttgtatgct aatgagatga tctatggctg 5820

ggggccctta gatagcttcg ggaagggggc tggtcgccag aaagactaat ttatgactag 5880

agggttagaa ctttcagccc tacctctccc acttctagag ggtagatggg ctacagattg 5940

agttcggtct ccaacagcca gtgaattaat caatcatgcc ttcacgataa accttcatga 6000

agacccctaa acaaccagat tcagagagct tccaagttgg tgaacatatt gaggtgctgg 6060

gaaggtggtg cacatggagt gggcatggaa gctttgccca tccactccca tgccttgccc 6120

tgtgcatctc ttcacttgct tgttccctag tcgtatgctt tataataagc cagtaaagtg 6180

ctttccaagt tctgtgactc gttctagcca attatcaatc ccaaggatag ggtcatggga 6240

acctccaatt tataactagt tggttgggaa tacaggtggc ccaggactct cgactggtgt 6300

ctgaagtggg ggcagtcttg tgcatttgag ccttctcact tgtgggatct gatgctaagt 6360

ccaggtagag aatgtcagaa tgcatctgaa ttgttggaca ccaagttggt gtccacacag 6420

aattggggaa ttggttgatg tgaggaaaaa ccctcccagc tacataggga aatcacggtt 6480

ttggaagatg agcagaaaag ggtgggaggg agaagacaag agagaagagg gtgaggaaag 6540

aggccaggca gcagtccaag tgtgagatga taacacctga actctggtaa ttgaggagac 6600

caaatccacg ggaaccagct gcaaaaggca ttctgtctgg taggtatagg gctaagggag 6660

taaacgtggg ggtgagactg ggatcaagtc cccttcctac cacttagaat ctgtggactg 6720

ggagcaggtt atttaatctc atgaaacctc catttccaca tgtgtgaggt atgtgattgt 6780

tgtctccacc cctagggtgc tgtggggttt acaagagata gtagtgcaca taatgtgctt 6840

agcacagggc cagcccctag taagtgccgg ctgtacttgc cttctctttg caggggtggc 6900

tcaggagagg gtgtaatcaa aagatgatta tgagggttca agcaccaggc aatgggagag 6960

tcagcaggtt ttaatgcttc cacacagagg ctggattctt ctacttccgc ttaccgctga 7020

cagctgaggc ctgctacctc aagccctgaa atgcagtctg aagacttttc ctggacttac 7080

tgggcaccat tggactgtac agggctgtgg cagacaggag cagtggaaag agatttggat 7140

gggacgtcag aacacccaga ttttacttct tggctcttcc acttactagc tgtgtgaatt 7200

tggcagtggt ttccttatct ataaaatggg ggaattggag agggtggtct actgcatgtt 7260

acttctcagc acctcagctc ctccccaccc ctctctcttg cttctaatct gcctacatca 7320

cttggtttgt ctctggacaa gtctaagttt gcatgacaac tctctaattt ctccttcaca 7380

gttatttatt tattttttgg cagctatttc cttgtggaga gatagcgtag gtctttcagg 7440

tgcttccatt tggccactgt ttgcaggtga agaggtttcc aggatgctcg cgggatgtta 7500

ctctggtagt aacagaaatg atggcctggg gaatgggaag agtatgagcc cagttttgag 7560

cctgagtctt tggggctcat ttattcattc aatctgcaaa tcttcattca gcattactac 7620

atgccaggct ctctgctaag tgctgaggat agcgatcagg acctgctagg gtgaaaatta 7680

ctcttcagaa gatagcagag tggataatgg ccattcgagt tcctcttaac tacacagagg 7740

gactgggtaa gatctataaa ggagagaggt taaacagcaa agcaaaagtg actgaaattc 7800

taacacccat gcatggggat tcttgagcta aagatctggt ctgtagaaga agacacatca 7860

gaccctcata tttagcatct aagccgaaat cagcagtaag catgtttctt ctcccataaa 7920

aacttgtccc taagaagaga gacaagtgct tacaaggact cttatgttaa aggaaactgt 7980

cacaattact ttatcttcag tcatgaagcc tttcttgagg aaagaaaaat tagcaatact 8040

agtttgggaa gcttgaaaga gagattctgt ttgcaatgac acttttaaaa aggcaacaac 8100

aaccaaaaaa gtaattattc ctaaagctca ggtgtcagaa gttgtgaatg tctagggggt 8160

tgtggatccc caacctcaga gacaggaaca gcaccactcc catcctgggg ctgtggcctc 8220

aggcctctgc ttcctctttt tcagtctaaa ttccagagca gagaggtgct ggatgaccag 8280

cttcaggcag gggagggagg aagtaaaggt agtttctcag ctcatcttgt cctcgtacta 8340

ttaatagcat ctgttgtttt cttaactgaa aattggagag aaccaaacaa atcaacatca 8400

tattggcggc tttctgcaca tcactcacaa atacatctga atgctctgta ctctccagtg 8460

tttattttcc tcagccacat aatcggaatt tgaacattct tttgacactt cttttccctc 8520

acttgaataa ttagttctat atataatcca aatattagct ggagccctct agcccctcag 8580

agatatagtt gaaccttcct ggacctcaat tagttgggca gttaattctg aatgttgatg 8640

tgcctatttt gcacgtatgg agagtttgta cgtcccccct ccccaaacaa gtagaatgct 8700

gtctatttgg tacttgctca ttcatacatt ctttcaacaa ccaaacccag ttgcagctgg 8760

gttgaacccc agggtgtttc ctgctattca gtgtgatgcc acattcacct gctgtggcca 8820

acattccttg gcccaggact gcagggatgg aagccaaccc aacatcagaa gtgggcaaca 8880

atgccagaaa cctatcaacc aaaacaaatg taatttgtaa tcctccctgg atgatgttga 8940

gtgattggca actgagcccc gtcatatagt aaatcaggct gtctctcttt aaaaaagaga 9000

gacagagaga gagagagtct cccaatgttg cctaggctgg tctcaaactc ctgggctcaa 9060

gcaatcatcc cttcttgcct cccaaaatgc tgggattaca ggcatgagcc accatgccca 9120

gcctaggctg tctctttagc tcttcaatta tagggcttcc tgcaacctca gcccttaacg 9180

aatgagctag cgtgtaagtg actagctaaa taaatgtgct gcaaacctct gacaaatggc 9240

tttttgacat tattaaaatt tatttatgtg gaacaaaaaa attcttttct tctgcatcat 9300

gcatcctaca ctgcttttaa agaaactctg ctatcctttg ttttaccgtc taacatttct 9360

ttgtttccct tctgacatgt ttcagcctct tgtcatcact tggaacattt ttactgcctc 9420

atccctctag tttccatttc ctagcctagc ccagcccagt atttttgaaa cccaaatacc 9480

tggaattttg cctgatgttt aaagccaaaa ccaaatgaaa atgtgaagaa ccaaaggaga 9540

ttttattctt ctgtgactca aaatcctgtt gttattgtta aatgaaccct gtgggataca 9600

gtctggtatt tttaacacct tctgggggta tttgtcaata tttacatttg atgtggatag 9660

cttataaaag tccagtggct ttgggtaaac cctgttcact aagaataaga cccagaaggt 9720

catatgaatc aacgatcaaa gaatgaagat attctgaatc caagtttctt gttttgatta 9780

ttaaaaatac cttgcaatta gttaggcatg agagaaagct tgtgtataaa gtttggcttg 9840

taattcacat gaattctcta tacaggccaa aaactacctt ccctttggga atatacaatg 9900

accttatttg gaaattactt caggtaatta tgaagtaata atatctctat tcatatgcta 9960

aaaagaattc aaaataacat tcaattatca tctcttagaa actaaattaa acgggtactc 10020

acccaagttg aaatgcaaac aaatgcatga tatcattatg tgaccctccc ctagccctta 10080

ccctgtccat ggcagatgct aagaaaaact tgtccctaag aagagagaag tcaattgatt 10140

tagtcagcct ttcccactgt gtttggactg agcctcagaa ccctccttat ccccatactg 10200

caaggagtta ctgtcaatca actagagatg gcgctcaaga taaaattatt ttccatcctg 10260

tactaatata ataataagca tggaatctat ttattggttt tttcacaaca ataaaatata 10320

tctttagcaa gtaacaaatg gtagaggtaa tacttttaga taaagaggga agaatccttc 10380

tgacttttct ctttatcaca ttttacccac acacaatgtg gatggaatct gcaactgaat 10440

ttagcagtca cactgaaact ctgctgcatg accacactgt cctgaggcca tgtgccaact 10500

tctatgtttt tttttcccaa agctcttcca aggaagagct gccacttctc catttactct 10560

tttcacctgt aaatgaatgc tgccgttggt attgagaaaa tgggtgaatg aaagaaatca 10620

tgagaacact cctggggtga gagcaagagg aatgaaaaca cgtggtgtga gagcaggtca 10680

gagatggtgg ttaacacagg aaccaggatg gaggcctcag acttgaccct ggcctgcagg 10740

acgttggcct gcattgtctg caggaggaaa cacagaggtc agtggatatc ctgcggtctt 10800

ccattaaggc ctcttactat tctggaacat tttattttta tagagaaatc tcccacttcc 10860

tgatctcccc agttagaatt gattgatcaa agcagtccct ctccctgtta ttaaataatt 10920

aaataatatg cattagttga tccctataac cacttaaaat gtatctaaca caggagaaat 10980

ctttggtaaa tatttactga atgaatgaac aaataaataa ttggagggag ggagaaagag 11040

acagagttaa atcctgaggc tggagttttt ggtccttcag ccccttatcc cattgcccct 11100

ggcagcccca tccatgtgac cttgaattct gtccaagtct catcactttc tatgtgtgtt 11160

gtaatggaaa aaaggttaca aaacacagtc tttctggcct ggagtaccaa aatagtaaag 11220

acttctgttt tgaaagcctc tgacaaatga acacaacagc cacagtgata actttagcag 11280

gaaattgaac cacaaaaagc aagcgagcca caaagccctg aaggctaaaa atgcctcaca 11340

ggccaagagg cagttaggtt gcacctgaga cagaggctgc tttcttaact gggaaatggg 11400

tgtctccaga tatataaagc tgtgaggaga tcaccatggc ccagctctga cgagccactg 11460

gtacagcatt gttggttagt ttgcctcaac ctgcagcaga gtttttattc caagagttaa 11520

cagttagtta ggctttttcc aaaggttgag atgacacccc aactcccaca cccatttccg 11580

tcctgcccca gcttgaaacc aggcaaagtg agttaagaca ttgggagatg ggaggggaca 11640

ggaggggtag aggcctggca cccctctctc ttctgtttca agagacactt tgtggagttg 11700

tctagagaga ctgggggtaa ctgctccttc tgcccccaat ctccccttag ctccccactc 11760

ctccctatcc tgccaggcca accaccagca gagaagggcg cttagctcca gtgacgctgt 11820

gagtgaggta cttgtgagga tgcatatggt tcctgttgga aatgcacgga ggagcagtga 11880

aacaccagaa gccccatcct ctctcctgcc cgtgccctca tgcccagcca gctgaatatt 11940

cctgtggctt ccactcctgc actgtcccct cagtgtcacc tccccaccac attcccacag 12000

ccccacttgc tgtgtcacct ctcccctgag ggagaggtgt cttcctgcca gtctcttcac 12060

caccaggctc acacaactcc catccattca ctgcaggctg ccagactgag ctttccaagc 12120

acccccggac tgcctccagc cctactttct cttcaaggct tcccacttcc ctcaaaatga 12180

aagccctgac tccttggaca ccatacagac cctccctgat ctcccagcga tgctcttagc 12240

agcatcattt ttttttctgc cagattttca ggtgccctca gttccaggag caccaaatca 12300

cctggatgcc atgagctgct tatgacacag ttcaaggtgt gacatctttg aggtctctcc 12360

ataacacctc tactgcagac gtctgtctct ggtgtatcag ctgtaccttg cacacatgtc 12420

tctaaccatc tgtccactgt gtctgtctgt tcacacatca gattgaccct atgttctctg 12480

tgacaccagg tgagctctgg caccagtgcc cagcagatga ttactgaatg attaataaaa 12540

ctactcactc cacctgctgc ttttaaatgg gctccatttt ggtatgattt ggtgcgtgaa 12600

ccattaaaac ttttcagaag tcaggatttg aaacaaactt ggtaaaaagt agttaagcaa 12660

tgtcttcgtt gtaaagtgag aaaccaagtt gcttggaaac aggtgaaaag cttagccagc 12720

agcttcacaa ttcacagcct ggcactccag tagccctctc tggacaattg cccaggtttt 12780

cttgttttca tttctgtgtt ttaatttaca agttaccttt atatttctac taatacctta 12840

ctggcacttg tttgtgaatg aaacaatttt tttgagagtt tgttaattgt tctgtgtgag 12900

tgtggaaagt ttctttcata taagatcatc aattcttttt ttaaaaatag tgttttaggt 12960

tttctcctac ataaagaaag tggtgggggt gaggagctgg gggtggacca gatgctcaca 13020

ttgtatttct atgattttgc tgttggagat actgaagatc attttgctgg gctttgctct 13080

cttaaattac agaaaagtgc atttggtgaa gtttagtgtt tttagtaaac atatacaaga 13140

tgtcataggg atagaattag tgaaggaata gggttcgtga atgaatgagc tcaatcttag 13200

taatgaaatg tttggaaagc aacagaacaa gagagaaaga gcatcagaac atgatttagg 13260

taccccatac atttatacaa ataaaaaaaa gatctgattt agggtcatag aaatcacata 13320

cctaaactga tggttaaaca ataagtggca agtaacagtt tagctacaaa aaatgtacag 13380

gaacacataa ggaaagttac atgggaactt tcagagagga ccaaaaaagg tggaataaga 13440

ttatctgctc tgttatgatc ccctgtattt ctctgctcta tgttcttctc ttgggcagtc 13500

ttttcctgag atgttttcat gttattaatc aaaaataata gaaacgaata aactagatca 13560

atgcatttac aagatttttt gagtgcctgc agtatgcctc ggtgtggtgg gaacttggtg 13620

tactgaggtg agtagcatga tcctgaagag cttaggaatc actgagttct cgtaagtgca 13680

atagaagaaa aactggtaga aggaagtcaa atgaaaatac aagtttctaa agaaaataag 13740

tatttacaaa ggaatttgtc tgcattaatt tctgtggaaa agttagctat gtatagaaga 13800

aagaaacttg ttagtaattc tttcattcat tcatttattc attcaacagt gaatccctgc 13860

catggtagta ctaactactt gcactgtggt agtgcaaggg atacatttgc tgtgcaagta 13920

gttaggaatt tttttttttt tttttttttt ttttagtgct ttgtgtatgc taggctttat 13980

gcaaatatac agccaagcac agaggttaag aactttggaa tcaaaccgaa gtggttctag 14040

tcccagctcc cccttacaaa ttgtgtgatc ttcaacagtt catttaactt ctctagatct 14100

cagattttct catcagtaaa atgagtgcag taagagtacc accctcacag catgaaatca 14160

gattttctgt actctttact aactcacttt ccatgacgct ggcatgtggc aagtgctgac 14220

tcaatactgg ctcatttcct taactgcttt tattttattc attcactgcc attccagagg 14280

aaactgcggt tctgagatgt gccctgggtc actgctgagg agcggacgga gcctgggagg 14340

gagctacccc tgccttgttt cctcacactt tatggggtgg gtgtcaggaa tgaggctgga 14400

ggggaaggca aaaccagatc cctaagagcc ccctggaccc tgcttagggt gtttacttct 14460

atcataaagg ctgtgcttct gtagagacct aaggcagaga gtgccaaggt caggtttgca 14520

tttgagaaaa gtttttctgg cagcaatata aagaatagtt tagcaaaaga aatgagaata 14580

gctggaaacc cagttaggag gctgtggtag tccaggagca cactaaggca gcaacaatag 14640

gtggatggaa actggagcat ttaggtaagg gagagggatg ccagaggacg tgatggtgag 14700

ggagagggaa gagggtggga ctccaccgca ccagggcttc tggctttatt gactgggtat 14760

atggtggcat cctgggccaa gacagtgagt cacagggaag aggctttgag cctggtaagc 14820

ttgggacact tttgagacat tgggaggaaa tgacacatgg cagttgacat acagatctag 14880

acgtcaggag aaaagtcagg gctggagatt ttcggtttac agttgaaggt atgagtagga 14940

atgagatcat ttactcagtc catcccacca cattgagcac catgaataag gggctcctct 15000

ccccatgggg caagagaggt gaaaagacag tcaaagtcac tactttcagg aggattaggc 15060

tcaagttagg atggggaggt ggagaaggca tcaaacaaat aaatgaaaac tataacttca 15120

gttattgcca agtgctagga atataatgtt gctgttaaac tgcaaggtgg ctaacaagtt 15180

ggatagtctt tcttgaagaa ctgtagaaac attcattaaa aacaaaataa ttttatagtt 15240

aagcaagaaa gtgattgtac tataacaagg gagacttatt tcaccatagg aaaaaaatca 15300

taacattcta agaatagaag gaaaaaataa caattaccca agcctgcaac aacctgtaag 15360

accagatcag cttttaagag gaaagttaat tagataacag gaagaggtaa aaatgagatc 15420

atcagagatg ggggaaaagg caaatttaga aattctgaat ttaatagggg agacaagtat 15480

gtaggaatat aaacacattg tgctttctct gcacaaacac attcacatga ggatgtatac 15540

acaacccttg caggcacagg tactcagtga atgcctgcgt gccaaggagg gggacagcac 15600

tgagcaggag aggtggcagg gcctggtggc ccaggagcac tcatatagtc aggagactga 15660

tataaaagaa agatcctgct aacatgccat tacagagtat ggaatgagca aggaaggaag 15720

agggtgaggc tgtaagagag tgcacaggcc ctcatttaaa tggacgggtc accagccagt 15780

acttcatgcc tgttcctaga tgacccacaa agtgaaacga aactggatgt ggtatttagc 15840

aagtctgaga tttgctggtt taagtactaa gttagcaaac cagccttggc agccagagca 15900

ggtagggtag ggggcccaag gagatgggtg tatccaggag gccaatggag gaaggagagg 15960

aaggttttct gggaagccag ctgacatgtc atgctttgtc agtggggcct ctgcatggat 16020

tccctccact cccccccagc ccccaactca ctacggggag ctgcaaaatg acagacaatg 16080

ggtggttttg tacccagcag ccactcctca accagtgagg aatgggagag agataagaag 16140

gtgggaaaca aggattgctt gtcaccatcc ctgctcctcc tgaagccctg cgctgggagc 16200

aggtacagaa cccgctgaag gtcacctaat catttcacca tggagttctt ccccagatgt 16260

gaaactgctg ggactggagc aaagggtgcc cagagctgac tggaggagga agccagagcc 16320

acactggaac atatggagat gctagctcct gcctgctatt gaggaacttc attttcccag 16380

aagaattaga ggaccacagg agtgcttggg aagaagcaat tgtggcaacc taggtgaatg 16440

atgacctagg aggtgaaatg aggaaggagg cctttggtct ctgccaacca tacatttcag 16500

gttgcctttt gctaaagtcc tgctgatgtt aatcattctc tccagcagat cgcggcttga 16560

atcatggggt ctgaaatgtg ggtttatgag agaaaaacaa catggaaaaa ttgggcagag 16620

gctggtaaat gtgaacattc aaacctgccc ccaatctgag atggtgggac agtggacccc 16680

tggccagctc tccaggtgtt gcgtatgcac attggcaaac cacctttatt aggtggtgtg 16740

acacctgggt agcaattctc aagcaatgtg atgatctgaa tgtccctggg cctgacccag 16800

gtgaggagga cacaaaatca tgcaatatgt ctcatagagc ttagtcagtt ttgagaaata 16860

ctggactaat gttatgtttt taaagcacct aattcttttt ctcttttctg ttttatctag 16920

gcactgggaa atcatgaatt tgataatggt gtggaaggac tgatcgagcc actcctcaaa 16980

gaggccaaat ttccaattct gagtgcaaac attaaagcaa aggggccact agcatctcaa 17040

atatcaggac tttatttgcc atataaagtt cttcctgttg gtgatgaagt tgtgggaatc 17100

gttggataca cttccaaaga aacccctttt ctctcaaatc caggtatttt ctacttttat 17160

agcactcaat gcttgaaaat agatgcccta aatcacagct tggcattata tttatggact 17220

gtagataaaa gtaggaacaa ctgggtaaat agtgcactag aataacactg ataaaagcaa 17280

tctggtaaat tctatgctgt tgttccaagg tacttcttaa agagatgtcc tcttctgctc 17340

agtgaacttc tttgttgaca aaaacagtat ctttgtctaa ttcagtgctt tcaaaatggg 17400

aagaggagac aatgtatagc cagtgctaaa cagtacagtt tcatttacta gagaatgaga 17460

gcctcaaaac tcactgccct tctcctgtgg agccatgagc acagagccag agttttacct 17520

caatttacct tattgctcct tttctaggat gattattaca tgtccagcta actcaccttt 17580

ctcgcttttc attattcatc tcgttttaat gacattcttg tatctgactt tctgggaaat 17640

atatatagtg gtcttatgat atatatggat atatataagt gtatgtatat acatttatag 17700

acatatatac acatatgtat atataatata tataatctta agacacacac atacatatat 17760

gacacatata catacataca tataagacac acacacatat ataatcttaa gacccttcaa 17820

gaaactattt tgaggcactt ttataagaaa actttttgga atattttggc aatgagaaca 17880

aaaatgacat tttctttaaa ttatccaaat taaatgtgtt tccatatttg aagaaaaaaa 17940

gggaaaatag gagccagggt tatctagaaa catgagtttt cattaagaac taactgataa 18000

aggacatata attgacaatt caaaaatata tatttactgt tgttgttgtt ataaaataaa 18060

taagtccagg ccattggaaa caacaacaga aaagtcagct gtgtttgctg agagcagcaa 18120

gcgggtcctg cagctggcat agaggcagag ctggaggcag ggtggagcgg ctggggctgg 18180

agtggtcagt gagaagagtg agagaacttg ctaaagaacc ccccaacccc accctggggt 18240

caggtttact tgctgaagaa cctccttggt ccccttgaga gctgtgtaaa acatccctgg 18300

ggcacaggac attccagaag tgaggtcaga gtcacaagct aggatgcagg ctccatgagg 18360

acagaaacct ctgctgtctc actattgccc aatgactgtg cacagtgcct ggcatatgat 18420

aagtcctcag gaagcattgc tgggatgaat acatgagagc aggaaggtat aggggccaca 18480

gtcacagaac agcagatagt tccatttcct ggagcatggg ttctgactag gaaaagctag 18540

aaagttggat cagggctaga ttgtagggct gagaagttca ggtgtgattt ggaggaaagc 18600

attgtctctt ctttaaaagg cacccataag gaggtaggag gtgcaggtct tggtggaaag 18660

gcacagcagc tccagaaatc aaactgcccc agctgggact ctgaccagcc ctttaccagg 18720

ctgtaacaga gaggggatca aaagtgacac tgctgtttcc agcagctgct ggaagagagg 18780

gctgcatttt gcagggccca gtgtgggggg aaagggatgg ggtggaatag aatgtttcca 18840

ggagcactgc ttcttccaag actacaggcc cagcggcctg gtgtgggttc ttcctacagt 18900

tctttgaccc taatcaagcc cagttctccc agtcctagtg gcctgggcac ccacagggtt 18960

ataacctgta tttctagcac atttaatcta atttatttta gctagatcat taaatacctg 19020

aaatacaact ctcctcatga catcttgaaa tataagccat gctttgtata tatgggatgc 19080

ctggtcctgt gaggtttctg atataataga ggaatacatt cctttgtcct cagaggcatt 19140

tctctgaatt ttttttcctt cttggcagcc ataccttaag caaaagatgg tagcatactc 19200

tcgacattta gtcattcccc tgggtacctg tctgtgtgcc aggcactgtt ttaggcacca 19260

gggatactga ccaaaaaaaa aaaagttgcc cactggagct tacatattca gtgggggaat 19320

caataaatga gtaaatggtt tggtatgtca gaaggtgcta aggctgtgga gaagaattaa 19380

gcaggagggg aatggcaagt gcagcagggg ggaaggtata ccttgttgtt tcaaagagat 19440

gctcaaggaa aggctcacat agaaggcaac atagcactga gggtcaacgg ggagccctgg 19500

gcttcttggg gcaaccaaca ggaagcagat tgtccttgat gttctcataa tagacaacag 19560

tggcaaaact acaactcttg atttttcaac attgcctatg agttacagac agccttgcct 19620

gtccaacaca tctgtgagct gctaattatt aggttgtcat tctggggcct ccctggctat 19680

gatgtcagcc atgaattatc ttccatggta atttgcactt gtggagttgg ctaatgttaa 19740

ggattcagga gttcccctgc tgaagcacac agacattcct aaacctcaca gtggaacaag 19800

ggatgcattt ctaataatct tgttactctt tggattagta cttttcaaac tggagtttgc 19860

agactcctgc attttgaaag cttttaatga aatgtcattc tcccaccctc cttcccttgt 19920

ttagtgttag ttcttagaaa tcagtgtaag aacgtctgga tctactggag gactccctag 19980

tcacagagta ccagacatga cttcagggca gcatgtctgt atttgcagtc atcttcatgc 20040

cagtgatagc cataaaacct caaagcttcc tcaaatgtgg ctaacagagc cagttggagg 20100

aaccagttgc cttgctgcac acagagaacc tgggttgcac agtgagagcc atgcttgact 20160

actgataaca aaaggctatc gtttgttacc actatgtgca gtaataagca cccagggaag 20220

aatgtacaca atcccagtgc cagcaacatt tagatgaggc cctcatgatt aacacagagc 20280

tcaataatgc tctaatgctg gcattattta tatatttata tattttttga ggcagagtct 20340

tgctcttgtt gcccaggctg gagtgcagtg gtatggtctc ggctcattgc agcctccgcc 20400

tggcaggttc aagacattct cctgcctcag ccttccaagt acctggcatg tgcctccacg 20460

cctgaataat ttttgtattt ttagtagaga cagggtttcg ccattttgcc tgggctggtc 20520

tcaaactcct ggcatcaagt gatctgcctg ctgcagcctc ccaaagtact gagattacaa 20580

gcgtgagcca ctgtgcctgg cctgctggca ttaaagagaa catgacaaag ttatcaactg 20640

ccatattaaa ataggcattc aaggactact tgctgacagt tttaagtttg aacactgatt 20700

atcccaagtt tccaacgaac gtcttggagt tgctaatact cttttgtgta tcatgcttac 20760

gtgcttaggc attttctgca ttggtattgt ggaagttgag atgtacaaat atataaaaca 20820

atatctgaag cctcctcaca gtttgcaacc aagttcaagc aaaacaatat ccacaacctt 20880

aaactaatac aacataaagt gaatgtaatg aggatattga tttgaatttt tatttacttc 20940

ttaggggttt tgtatttaat gaataaatat gttttggttt tactgactct tgagctttag 21000

taaattaagg tgtttaacct ttgcatgtta atatgtatat taagtaatat aataaaaatt 21060

gttaataaat atccatttta tttattttgt tccttaggga caaatttagt gtttgaagat 21120

gaaatcactg cattacaacc tgaagtagat aagttaaaaa ctctaaatgt gaacaaaatt 21180

attgcactgg gacattcggg ttttgaaatg gataaactca tcgctcagaa agtgaggggt 21240

gtggacgtcg tggtgggagg acactccaac acatttcttt acacaggtaa ttgtttcaaa 21300

aggattgcat gggccaggat gtccagataa gcactgtgtc tcttttgcct ttgtaactgt 21360

tattactctt tttactgcta tttaatatgt aatgtatatt atatgatcta taatatatat 21420

gtaatataca ttaaatggga acatgtgcaa atcttacaaa cattactaag aatttattga 21480

tttcatgatt ataagtcaat tccatatatt tatgtatata ataaatatat aagacataaa 21540

atatgtataa aacatatata taccatacat atgaaaaaca aaaagactat tgagccccaa 21600

ccaagtcatt tatgatagat tcttatcaaa aagcaacatg tcatataaaa actttaagca 21660

taaacaaaat aatgtaaata aggaccttta taactcaagg tggatttgat ttgtaggttg 21720

cacaggtagc agaggctgga cctcgagtgt gagtcctaac tctgctgcag actaggaata 21780

taagtatcct ggaagccaca tgaccctggg tcctcaggag agcatgcact cttcctaccc 21840

actacctgac tacccccacc actctctttt tttctctctc aaattggtgt taagtagttt 21900

tcatcagaat aactctttta tgatttttcc actttaaacc tataatgtcc caaaactttt 21960

gaatctggtt ccacctttcc cacttccccc accttgcact gcatagatct ttgaagtatt 22020

cccaaggaaa ggaagaagag gctggtaaac agttcttcta tcagcattcg caaagaaggc 22080

tttggcgtgg gaacagccct ccccttgaag aggctcctgc ttctaccatc cttatgtggg 22140

ctgtcaaaga ccatgctatt ttgataaaag cgaaactcaa aataccccaa acaataagta 22200

ggaaataccc aaaaacatta aaagggatta tcactaagaa agtaattttt attttctttc 22260

tagtactttg gagtattttc aaataatgaa tgcatgttac ttttgcaatc ggaaagcaaa 22320

atataaatta taaaaaggaa agaaacacaa tattgggtca aaagtcccca ttcctaattc 22380

acttgaatga ctttgagcag gtcacttagc acctctgaat ctgctagctt agcttcaaaa 22440

taggggatga tcatgctcaa agttccacgt tactgtgaaa attatatgag atgtaaaaat 22500

atgtgaaagc agtttataaa ctataaatag ctatattctg gtatgcagcg tcatggaggt 22560

tgcctgctta taaaaatatg tcagtcaaaa tcataactca caaactaaag ttggccttca 22620

cttaagaaaa taccagtatg aatgtgtgct aattaaattc cttcacagat attttacttc 22680

ttggtaaaca caggctaagg aataatttca aactgagtta aactgacagg ttgtgaaatc 22740

aatttatggg ttatgaccag tagtagggct tttttttttg acagataaga ctaaaataga 22800

ttaggcactt acattactca taagaatgat aaatattgtt ccatgggaaa agatgtatat 22860

aagatgtcaa gaacagcccc aggcacatag cagacaatcc ataaatgtga tgaggatgac 22920

gactagcact ggacattgtt agcatggata acattgcaat caaagagttt caacaactcc 22980

atgagggtta aatgaactaa gagagccaag acgaggaaat aattggagca ggtcactggg 23040

aagcactggg taggggtgaa gactcaacaa agtccttctg actctaccac taatggtggg 23100

gtcctttcaa tgcctgatta ccctcatctg tggaatgaag gtgataacaa cagtacctat 23160

tataaggggt cactgtgatt taatatgtgt aatgaatgta taggtaaatg gccttagcac 23220

agggcctagt atgttagctg ttcttatttt agttatgtgc tggctcactg gataagacct 23280

gtgaaaaccc aaaccttagc ttttcactga ggcagtgtca gcacccttgg acacacagtg 23340

ggagctagat atgtctcctc ccccttcttc ctttagaata atggtttcaa aaagtttaaa 23400

ctatgacccg tagtaagaag aacattttag acagggagcc aaaacacaca aacatatatg 23460

tgtgtgtgcg ttcacatacg aacatgcaaa agcacttgta tgtgcaaaca catacaagtg 23520

aaagaaaact ttcacaaagc acatggaatg caccccaact tttctgtttt actgttttat 23580

tcacttaatg ccagctacca ctcattgaat tgattttctt gtactgtagt taatccatgt 23640

ttctaaaaac tcagctttaa atgggacgtg accaaaataa tgcatgaaca ctcaaaggag 23700

tttcatccct tcattcagat gtgaacattc ctgtgttccc cttgacttgt ctttacacca 23760

tcttaatcaa tgctgtctta cagcaacaat tagtcattca tagcctgtgg tttgtgtcca 23820

tcagtgagtg tgccctgata gacctgaaaa gattgggaga ccatgcccag cttcccatta 23880

tttctgtagc ctgtcctttg ttcccataac agtacagtca tgccttagta tctgtggggg 23940

attggttcta gaactccctg tggataccaa aatccatgga tgctcaagtc ccttatataa 24000

atggtgtagc atttgcatat gacctgtgca catcctcgca tatattttaa gtcatctcta 24060

gattacttat aatacttgat acaatgtaaa tgatatgtaa atagttgtta tactgtattg 24120

tttagggagc aaagacaaga aaaaatatat gttcagtaca gataaaacca tcctttttta 24180

gaaaataact ttttgttcat ggctggctga atccaacaat gcaaaaccca cagttagaga 24240

cagccaactg tatgtgcctt tgctaagtag ataaatttag ttaatgatga aggactggat 24300

gaaagaaaga tttgaggata aaaattagag tttgcttatc tgtaaatcct gaagtaaata 24360

attacagtct ctgagaaaac tgagataact aatacttagt catttacttc acatttcatg 24420

attgcttacc aagattaaac ttgttctgat gaaaatcttc acttttcata ctcatcgttt 24480

tgatgagtga acttaacaat taaaggtggc tgggcacagc agctcatgcc tataatccca 24540

gaattttggg aggctgaggc agaagaattg cttgagccca ggagttcgag accggcctgg 24600

gcaagatagg gagaccccca tatctacaaa aaaaattttt aaaaattagc caagtgtagg 24660

ggcatatgcc tgtagtccca cctactcagg aggctgaggt gggagaattg cttgaatcgg 24720

ggaggttgag gatgcagtga gctatgatca ctccactgaa ctccagtctg agtgacagag 24780

tgaaaaccct gtctcaacaa cagcaacaac agcaaaaaca ttaaaggtaa ttccaggaca 24840

caatagggat gaggcatatt gcctgtttaa ataaactttc ctggccttaa gaaaacaact 24900

ctgctgaata attttatgga attagaatga cattaaaata atgaatctga taaatatgat 24960

tagttaataa aaatacagtt aaaataatac atcccttgga attttaaatc ttactgagaa 25020

ccatttctgc acgcacagta tccaatatac aaacacacat ggaagaaaac ctttccaaag 25080

ctcgtggatt gcaccctcat attttctgtt ctgttatttg agtttttaaa tgctggttac 25140

cacccattat attaatttta ttgtattgta gcttgctcat atttttttaa aactcagctt 25200

caaataaggc atgactaaaa cagtgcatga atggatccct tccaaatgtt gaaagttatt 25260

ccagtttctt tcctttccaa gtaccatgca tatgcagcta accctagcat tggctgacca 25320

gtctaacatc agagtgaaga gaagggaact tccctctaca aggcttacaa aatgataaag 25380

ttacaaagat aagagctaaa atgcaaacgc tttctctaaa gaagttaaat cctagagtca 25440

cgaactggtc catgttgctt cctttagaga aaaaagtagt aaaatcagtg caggttgaaa 25500

cctgcagcag gtacaacagg ggactctatt ctctgtgctg tccttgaagg gactccaaag 25560

atgctgtttc tctgtttgcc tcccagtatt ttaagtgagt tttggattct gcaaatcaat 25620

ctggctagct atggccttgg gtgaatataa gcaccagttt ctatagccgc aagttaaagg 25680

gatttgaatt gttaattctg tcatccctcc cattctaaca tcctgttacc tagaaaatca 25740

tccatacata gagcctgaaa gcttactttt caggatttgg ggaaataaaa aaaaatttgg 25800

tagaatacct catactttca ttaggctact gatatgaact taattattcc tctaactcct 25860

ttgaggctca gaccttctgt cccctgtctt atcactctca tctaatcacc tccaagcaat 25920

gtcccacgtc actgagaatt tagtatccaa tttcctatct tctgctccac tccagctcct 25980

gccttcacct ggcagagtgt ccatgtgggt agttgatcca ataccccaag ttcagagtcc 26040

cttgatctcc tcaatccagt gactgacacc tccactccat ttcagcaact tcacttctgg 26100

ggccacaggc tgagacttga ttatctccag aatttctcca tgtccaaaac tgaaatggga 26160

tagttccctt gaccccttcg tgggactcat gaaggggttg tcttgtttac tcatcctgca 26220

gctctcaact cctcatggga gggagagcac ataggtgagt gggtgcagag ggtgggacaa 26280

gtgcttctgg gtgctggcag gagtagaact ctgtgcggcc ccgcagcagc atctaggtga 26340

gtatctgtga cccctggagc cgcagagggc atgtgttaca gtgtgctctt ttagctttgc 26400

tgtccatgga cagcttaagt gttttaacag ctcagtgtga cagccctctg tatcctgaac 26460

tcttgttcag caaccaggaa ggatcagctt gcacgaacaa attgaagatg gtgaatgtgg 26520

agaactttat tgctgatgaa aatggctctc agtgggatgg ggagctggaa aggggatgga 26580

gtgggaaggt gtctccccct ggagttaggc catccccagc tgaactcttc tccgaggtcc 26640

taccgtcaag ccatccctct gaagtcaagc tgcttttcac cagtgtcaag cttcttctct 26700

tctctccttc tctgccattc cactgccagt ggatcctggg gtttttatgg ggacaggatg 26760

gggggcaggg cgggccaggg tgattttgga aaaggcaaca ttcaggcagc aaaatgggga 26820

tataaagttc tcactttggg ccacgggtcc aggcttgagg gtggagccat tgccagggac 26880

cccacccttt tctacctagt atttccctgc ctccactctc caactctctt atcacaggcc 26940

tcctatccat aaatctttct cactccctta ttttttttag agctcacctt cagacttatg 27000

aaagctcaag tcttttgaat tcctctttcc ctcccagtcc agcaggcttc acttctctcc 27060

ttgctagcct ggaagtgtcc attacaccct tgactgtgtc gcggctgaca catggaaggc 27120

aggtgtctgg tattgttaaa tgaacgttaa caagtgcaac ttctttaaaa agtaagattt 27180

ctttatttga ttgatcagtt ttttttttaa ttaagtatct aatttccatt gcttctggat 27240

ttaggcacac atattgaata taaaattcaa aacttttcat gtatctaaag aaataatttt 27300

aatgaccaca gaattttgct cttaaaacaa atggagcttg caatataatt ataactaaga 27360

cagactacaa gcacctgctg tgttttcttc atgctgccat caggctggtg catagtctga 27420

tgggatggat gcatccctgc tgtaatagga ataagtaaag agggtgggag actgttgttg 27480

gtgtttacaa tcattattca tctctccatt aattttcatc cttgcagttg agataatctt 27540

gccttttaga acgagaaaac aatcttatta cttacaacat tcacaagaca gggaaacact 27600

cctcaacaaa gaaccatttc ctttgggcat ttgataagaa ctgctgacat ctgtgtttat 27660

taagtgatgt gggagaatca gccaaggggc agaggtgtgg gtggtagctt caaaggtaat 27720

ttcctgactg agtgtggttg ggcatgttgg caatcccagt gtttgcaatt ccagtgcttt 27780

gggaggctga ggcaggagca tcacttgagc ccaggagttt gaggctgcag tgagctatga 27840

ttacgccact gcactccagc cagtgtgaca gagagagacc ttgtctcaaa aaaaaaaaaa 27900

aaaaggattt tgctttcctg gtctgtttga atgtggtaag cacaagaact tatttgtact 27960

ggtggtcacc cacttgggtc ctttcagtaa acttctggcc atggcccagc ctgagcacct 28020

gcttccttta cagagcaaag ccagagcctt ctggtttgta atttttgcag ccacataaca 28080

ctgatccatt ttagctgtct catgaaactg ctttccaaga tgaagtgaga ctatccgaaa 28140

ccaaaatcta aaaagaatga ctacagaacg aatggcagtc caaggtcatt tggcagggta 28200

gaattataag gaaaggtgat ggccagctac tgtgaagtct taagcccact cataggtacg 28260

aatctcttac caggactgaa gtttgtgttt ccttaacttt gctgcttgat ggtcttctgc 28320

aaaggatctc atttctccag ctttcactgg gaagtggcat ggggttttcg aaagtgcttt 28380

aaactctaag aactggtttc ctctcaggct tagccaccac tagctgtctg actttggaca 28440

agttactcaa ccactctgag catcagcttt ctttttttaa aatggtgaca gtgatatggc 28500

taccttagag agttgtggag attaagtgag ataatatatg aacaaaatat ataaatatgt 28560

ctatattaga catatatgat ctgttaaata tatttatata tatatataga aagaatatta 28620

gagaactgac atatttatct tagacctact atgttctagg gcccagttct gctcatgtat 28680

tatctctcta tatatatagt attgagagag agaaagagag agagagctag agaaatagaa 28740

accagtagga tatatcctat ctgaagaact ctaatacagg tatttcactg aacaaaatgg 28800

cttctcctta aaatgtattc caacaccaat ttcccctcct tgcttctcat acatattaat 28860

tcacagaaat tatattggtc ataaggcggt aggactatga ataatttttt tcttctgcat 28920

ttgccaaata ttctgtaatg ctagttttat ttttataaca aatttttaaa gtccctgtaa 28980

accctataaa attaactact aatcctaaac ttgaactgta atacatattt ttgttaaata 29040

aaaaaaaaga tatcctaaat ttgagaatct aagctaaatt cacaaaatga ctaaatgtat 29100

cttcttactc ttaaacgata agaaaccagg aatgcagtgg tttaacccaa acagcctagg 29160

tattcttcac tcatttctct tgtcctcagt atatgtttcc cccaggccaa gaaacattta 29220

tggctaggaa ttcatgactg ctcatctgta agacactctc caagtactct gtcatgattt 29280

ggaaccagag ctttggcctt ccattccaga tgacctccag gttctaagct ttatctgaag 29340

tactccacac tacattgcac accagcatga tgggaaggaa tgactcaagg gaactgggca 29400

ttcagaccac tcaccaacta cacacagcct ttcacacagc agtagacaac ctttgtagga 29460

agacctctgg ccaaatattt atatatcagg agtatatcaa tgtatggtca cagttacctc 29520

ctttttaatc tcaagaaata aagtcattac ttctgaaaag aacaccagta atggaactac 29580

acgggcatgc caagatgaaa attagtctct ctctctctct ccaccccatt cttttggcca 29640

ctgcatttta aacaggccat agtgatagaa ccagaaggta gctgtgtttc gattattttt 29700

gttttcttat ttttaaacat tgctttatag aagcctgttc tataatgtaa gaagtgaata 29760

ggaatcctag tatctggaat gatgctgagc catggagatg gccagattcc atgtgaagag 29820

aagaggctgg cagaaagcta agtatttgct cgctttccct accctgtatt catggagcag 29880

ccattacaaa agatgacgca ccagaccatg ggcctggtct tgctaggtgg tggagatcca 29940

aaatgctggg cactcactgg agctccgtga agatctgttg aatgagtgac taaatgtgtg 30000

gtgaactgtc cctacctaca gggttccctg ctttgcccag cctatcaggc ttgggtacaa 30060

gcagatagcc tggattggat ttcatgaggg ctttgtgccc actctgtata ctaccggaag 30120

atgcataatc tctttagact tcctgggtca tcactgaaag gccagaggca agaatttgag 30180

cagtaaattg aagcatgtgc agagtgacat agtctgggca tagataacag tacctgaccc 30240

ctcaaattcc ctaaagttga catgacagct tgtgcatcat ggtttggtac agagatgact 30300

gaagagacag tggttgctcc tctggttgct ggacaccttt tgaatgggct gtgatgactg 30360

gagagcgtgc tctgtcctgg tcacctgaag gtgctcctag actagcagaa gcagagacag 30420

cagccaggcc tgactggaga gctggaggga ctgggtgcag gggacttcta ggccagcagt 30480

ttctgtgctg tgctcggagg ggctctggtg gcaggaacct agtcctgggt tccagctcta 30540

ccagggtgca gcctgaggaa ctcagctttc ctatttattg gcttgccttg ccacaaagga 30600

tttcttgtgt gttaaagata ctaatagctt aaacaaacaa acaaacaaac aaacaaacta 30660

ttgatttaaa ccactaatca aatctttggt tttacaaatt tagaaaataa aattaggccc 30720

cagggactaa atgtctctcc tatggtccag aaaccagact gaaactgacc tagtagcaaa 30780

acccattctc ctgtatccca actcccagtt caggttcctc ttcattagct cccacccagc 30840

tgcatagttg aggacactga attgagggga cctggggtaa agggctggca ctcaggcatc 30900

gctggcttct acatgattcc accacatgga aagcatcata atcatgtatg atccctagaa 30960

aagtgtttgc taaactcctt aagttgtcat tcaacaaaca tttatctagc acctaccaca 31020

tgctaggcaa attctagaga cacagctgcc cctgccctca tggaactaaa tgtcttgaaa 31080

gagatactgg ccctgatcag attttttaaa aatacataat tatgcaaata aatcatatta 31140

tatataatta caaagcatgg tatgaaagga aaagtacaag atgctgtgaa agcgtataaa 31200

aggaggactt ggccaagtct agaggatcat cagtgagtgt tccctgatgg actaacatta 31260

gggccaagat cagaaagatg tccaggagtt aacatggtca ataatcactg ttaatcttta 31320

ttaagcattt cccttgagtc gagtcaggct tcacttgctg cattaacttc tttaatctcc 31380

acaataactc tgtaaggtag gtgcttttgc tttttttcca ctttacagat gaataaacaa 31440

gaagtaaaaa gatgtaacta attcacatgt agccctctgg cttcagaagc tctttactcc 31500

ataaggttcc tgggcacgtg ggtgtggtct aggaatggga agaggctaag aatgctgaag 31560

ggcagactga atgttccaga tgaagctggc gtgataagaa agggccacat catgtgggtc 31620

ttataggtta tgatattcat gttaaaactg ttaatgtttt tgaaatatct aaaaccagga 31680

agtgcatgat catatttatg ttttagaaag atcactcagg ctgctgtgtg gagactggag 31740

tagaaagaag catagaagag acctacagag actaggaagc tgtcacagta acctaggcac 31800

aagatagtag tggtcaagac tagcttgctt gaggaagtgt agagaagtag acagataaaa 31860

ggagcatcca aggagtgtac acttctaagt aatttaaaag aaagagtcaa atcccttaga 31920

agcaatcatt tgaaataacc taattaccaa tcctttttat ccagtcctct tatgttgtcc 31980

caattcagga attcttaaac cagttcttgt gttaaaatca tagcatggac tgccttacaa 32040

cattctatac ttcagtccat tgaaaaccac atcaggccag gtgcgtggct catgcctata 32100

agcccagcac ttttggaggc caaggcgaga ggactgcttg agcccaggaa ttggagacca 32160

ctgaactcta gcctgggcaa cagaatgaga ccctgttgaa agagagaaaa ggaagaaaga 32220

aagaaaaaag aggaaggaag gagggaaaaa aggaaaaaag ggagggaggg aaggtggaag 32280

aaagaaaaaa aaagaaagag aaagaaaagg ctgcttcctg gcacttctat taccatgtga 32340

cacgagaagg gcaattgtcc agacccacac ttaggttcac cacagtcctt ggttcccagg 32400

acagagtcct ttgaggttcc ttcaagcacc cacaccatgc tggaaaatgc aggctttata 32460

aactccagat gtcagtccac aactccacag ccccattggg tttggttcct ggatgaattc 32520

tcggctagat gactggaata gagttctggt gaacatcttc cttcctgcca agatgacaag 32580

tcaggtttct aaagacagat gtgtagctct ttgtggaaaa aatattctgg gcacgaacat 32640

cttcggctga acggcatgga atggtgtgga aaatgtgaag ccctggactg gtttctgtgt 32700

cagtggtttc ctgggactgc cgagttgaag gggagcagaa agggctcagc tgtggaggag 32760

gcagagatta gccaaactag accagctatc gccccagctg catggatagc atcttccttc 32820

ttgttcctga ttaagctcat ttaatatgcc aatgacaaaa ggatcatgaa ggtctttcca 32880

aatgaatgaa tgttttctgg agctgagatt gctggctcct atttgcatag tagtgcatac 32940

tgaggcatct gaccggatta ctggattcct aacttttaca ttctttttaa gctcagatat 33000

ttctacggaa ggcagaaaga taaaggccaa aaaaagatca ggcttgccta gctcttgggt 33060

ttccaggaaa ctgacagact cctgatatct gcaactagtg ctttctctcc ctgggcgggg 33120

cgttctccgc caccttgtgg acatttgggg aactgcaccg ccccagcatc tatgatcaga 33180

ggaactgaca gaggctgcca taactacctg gtcccctgaa ataaaaacca ggcaccaggt 33240

ttgcccaaag gaggaccaac acatagcaga atgaggctga gtaggatgtt aaccttccat 33300

gctggtcttc cattatcacg ttatctgcca ctgtcacaga caacatctgc atccgctaca 33360

gatggagatg gaacaaaggc tttcactgga tctcaagttg gatgcttctg taaaagtaaa 33420

gttccaagac ctttttcttt tttttcccca gtgacagcaa gaaaattgga tgttactatt 33480

ggcaacaact atttcctgct tgtgaagcgt tggtatttgc agtgcttctg ccctgtggtt 33540

attgagctga aaggggggct gctcttgaca taggagcatg gcaggattgg attcttcata 33600

acatgctacc cagcttgtct tatgtgaaaa gagaagccca gaaagagctt gggacacaaa 33660

gatgcaaatc ctgggtttgc atttgatttg ttaagtgtcc tttcacaacc agcttgcaga 33720

ctttagggga aaaaaaatac atggtaggct tgtgggtagt ggagaaaaca taagctccag 33780

tacctcacag cctgggtttt cagtctgcct cctcccaacc ctcatcacat accagctgtg 33840

tgaccgtggg tactctgtct atgacaatca attttcttgt ctgcagagta cagataatag 33900

aggacctgcc ccatgggaag cttggaggac taaatgagat taagcccagg gcctggctca 33960

tggccgattc tcgtaacagg tccatgtgtg taatcaacat gtgctgaagt tctctggacc 34020

ctctgtaaat atctaatagg aacgactagc catgggtgtg atgagggaga tgctagaaac 34080

agtaatgggg tcaaatgaat ctgtcccata gtctctaatt tgaaacagct gaactgtgcc 34140

attgtctcct tggaggtaag atataagcca cagagtgcct gaagcaggtc tacagttaag 34200

gaacttaaat gtgctaagtg attaaaggct gggctgccta ctccctgtat ggcgacactc 34260

tccaccctcc tgcatactct ccactactcc aggcagtggt gggcctccta gcctctgggt 34320

gatgagcagc tcccacatct cccctttgag acagaggcct gcagaacctg accagtgtga 34380

gggctggctg gggtcaggag tgtgcctgct ggttggccct caaggtgaat gtggctcttc 34440

taaaagacac ctgaagggat gcagaattta gcagcaaacg ccaagaacag ctctttgtca 34500

tctctgcatc ttctactaaa gattttaagc tccgaagtat atgtctgggg tttctagaga 34560

ttgctattcc cttataaggg aaaaaataca cccttattgt atatttattg agcacctaca 34620

atgtgccagg caccccagct agggagttta gatatttaat cctctcagct gccctgtaaa 34680

atagggattg tttaaaatgg atttcgccca ccttataggc aaggaaactg aactacagta 34740

tggtaaagtg gcttggctaa tccacctggc tagtgagaat ggagccacag tgaacctgga 34800

ggatactctt cacggcgttc catctttgtg tgccccacag cacaagcaca atgcccgggt 34860

cttaggaaga gcttggtaaa tgactgctga gttgagaatg gtaggattca tgtaccaaaa 34920

gggcatttgg aaatgggcaa atgcccagat agggctctga aagactagct atgtagagca 34980

acagatggca aatcatcaat ttaaaactct gtcatccagc cagtagcccg ctggcctaca 35040

gccagccatg tatgtacaag ggctgacttg atcagctgtg gctcttttat ttcaggcaat 35100

ccaccttcca aagaggtgcc tgctgggaag tacccattca tagtcacttc tgatgatggg 35160

cggaaggttc ctgtagtcca ggcctatgct tttggcaaat acctaggcta tctgaagatc 35220

gagtttgatg aaagaggaaa cgtcatctct tcccatggaa atcccattct tctaaacagc 35280

agcattcctg aaggtaagtg aagttcaggg gaatgttcca ccaatctaaa atttagatgg 35340

ctggatattt tgctccttcc catttttttc ctttaatgtt tcaggaaaag actataatac 35400

tgttgaagaa tttagtttct tccttgagtt tgccctcttc atggatagat ttaaaacaca 35460

cccaaatgcc aaaagcccca cttgcttctt caagtcttct cagcatcaga ttccatacat 35520

cactgcttcc tctgcctttt aaaatcttga tcctgacatg ccaaagccac tctcagcttg 35580

tgtttacagg agagataact gaactaaagg ggttctaatt gggcagagcg gggctcaagt 35640

cctggccccc caactagagg cagtgtgcct taggcaagtt gcccaatctc atgggcctca 35700

gtttctttat caaatgaggg agtaccttat ttatatgtct gagtctattg cgggatctgg 35760

ccagcagccc gcaatgcaat ggggctctct ctttgttcct aggcagattg gcaggttgag 35820

aaataataga cacacacaag atagtgaaag ctgggtccag gggggtcact gccttctggt 35880

cctgcggtgc caacaatgca ctggatatac cagcatttat tattaagttt agtgagggca 35940

ggggtacgtt agtgagggat ttagggtcat ttgattatga ggtgagatgg tcacatgggg 36000

atgaagtaat tctttaacat aacatttgta tgtagaagta cagtacattt gtatgtagaa 36060

gtacagtatg agataagaat ttacaatata gtgtgtgcgt cagtaatttc taacagagcc 36120

ttagaacaga aacacagtct ttccataacc tatgattagc aagatattaa tcagcagtaa 36180

caattgcaac aaaaactggt tacaaataat ccatggaaac aggacatgaa gctagataac 36240

cagttagacc agaaattctc agaggggagt acgccttaac cctaaagagg cctagaagag 36300

cggcggcaag atgagcgtgt ttatagccct atcttatcca tatggacagg ccccccccat 36360

gcgtccgttt ataggctccc cacaagggtc acattccatt cccagagcta tgaacatctg 36420

cttttctggg ataggaatct tggtgatggg aaacctccct gactgcacgt ccatttgtag 36480

gctctctgca gggggaagca catcacgtgc tgttggctca ttgtggcagt ccaacctggc 36540

attgtcttta cacaatcctg catgcaattt tgtatttaca ataatcagga gcatttcatc 36600

ttttattccg tagcaatagt ttcagggcgt ctccctacat gagtctcaga gtaaaaatac 36660

atttttatat cattgatttt catatgtgta cagaacacac tcctgtgctt gtctctcact 36720

ttcccttttc tgttgactat gatcatgtca ggaagatgag gtgcacagca cccaggtgag 36780

cctgtagtca caacacagcc ctctcagtag ccagatggga aaggggccca ggaccacact 36840

gacagcgata gcccagctgt acctcagcag gccttttata actgcccctg ggcctttcag 36900

agctcctacc gccaccacta atgctgtccc aggtgggcca atgggtagct catacagatg 36960

tgatgagctg gtagatgttc agaatattct taaattaaga tgcattccag gtcttattac 37020

caggaaacaa aatagaagtt cctaaggaaa tttcatggtc aaaaccaaaa tgcacacaaa 37080

tatggtaaac aaatatgttc tcacagcaag taagatagat gatgtttcat tccttttcca 37140

gaatttagcc cagtgtgaga tttaattgta gggtaccttc ttttctttct tctagatcca 37200

agcataaaag cagacattaa caaatggagg ataaaattgg ataattattc tacccaggaa 37260

ttagggaaaa caattgtcta tctggatggc tcctctcaat catgccgctt tagagaatgc 37320

aacatgggca acctgatttg tgatgcaatg gtaagtcatc agcaggagtg gacatatgct 37380

agggaggaag gaaaggaaga gggaagagga aggaaggatg cgagaaggga tagatcgata 37440

gctacagaat gagggatttc aaaacatttt tagccagggt ggtggcatgt gcctatagtc 37500

ccagctactc gggaggctta ggtgggaaga tggcttgagc ccaggaggct gcagtgagct 37560

ataattgcac cagtgcactc cagcctgggc aaaagagcga gactctgtct ctaaaattaa 37620

ataaatatga aaaataaata aaacattagt gcctaaacaa aagcctgatg tagcttcata 37680

ctcagtacct tcttaaggtg ctggaaggag atcaggtaac caccaacata ctgctttaca 37740

agaactaaaa cctttatttt aaggtgttct tagatgtctg aattttggat gttgctttcc 37800

ttttgtaaga tgatgcaaat gcacaaactc tgctaccctg gagatatcaa cataatagat 37860

aaaagcatat ctgtttggcc agactgtttt tcactcatct gttcacttga ctccagagaa 37920

ccctggcctt gtgtgacagg aacctctttc acaggcacag aaagacaaca cacagtttta 37980

atggtaataa caagtcctat ggctgagcaa gaccaggtat gaagaacaag cccgtctact 38040

tgtcaccagg cggaatggcc atggtagctc tctgggggcc ttactgagcc aaggtctcct 38100

atgtcggact actacttcca tgggctcaca gcctgcaccc agggaagtca tttctgtcag 38160

tgcttggcat gcattgccca gtgcccagga tacaactgca ctcgataaat gtttgccgtg 38220

taaatgacaa caatcatgct catgccaaga ccagactcat gatgtctcct cagcatcatg 38280

cagtcaatgc cagagaagta gttaaggaga agctggtata taaatgttaa gagaagtctg 38340

gagggaaaca acccaggctg ttggcagtgc ttatccactc tgttaaagag gctaggacaa 38400

aggagagctt ttgcttttta tttatttatt tatttaatta attaatttat ttatttattg 38460

agactgagtc ttgctctgtc gcccaggctg gagtgcagtg gcatgatctc agctcactgc 38520

aacctccgtc ccttgggttc aagcgattct cctgcttcag cctcccaagt agctgggatt 38580

agaggtgtcc actaccacac ccagctaatt tttgtatctt tagtagagac ggggtttcac 38640

catgttggct agactggtct cgaactcctg accttaggtg atccacctgc cacagcctcc 38700

caaagtgctg ggatttacag gtgtgagcca ctatgcctgg cctttttttt tttttttttt 38760

cggctttttg tctcctaaat gagaatatat tcattttatt tcttataact aaacatatat 38820

tttgtaaaat aaaagtaaga atggatgtga ttctccccag cttcaaaagg ctgtctctgt 38880

ctttccaaaa ttgccaactt gggaatcagc tgcagacata gacgtggagg ccaggttctc 38940

tttggcatga ggtctggtgt tgagcaggct ctagggccaa gccctcagaa tcaggcagag 39000

ggcatggcct gccctggaga agcctgtgca ggagtgactc tgcaggccac cttgcatggg 39060

aggcccagtc ccgacaccag ggctacaacc gagagaccat tttgctaagg ccacctggcc 39120

tgacagtatg ggagttttag gagggggaaa taagagtttg acatggactg gtaacagtat 39180

atttttactt ctgggagcat aaaattagtt ctaattatag ttgagagatc cctgggaaca 39240

cagcagcagg tgaaaagcag ctttatttcc tcactagagc taaccagatt cctggagctg 39300

atcctaagga agaagagcca gagtaactag tgtaaatctg tgctactgtt gcagattaac 39360

aacaacctga gacacacgga tgaaatgttc tggaaccacg tatccatgtg cattttaaat 39420

ggaggtggta tccggtcgcc cattgatgaa cgcaacaatg gtatgctccc aggcccagct 39480

cctcagtgtg tcatgttctc tctctgatct ccttttctgt tatggttctt gccctgaaca 39540

cacccatgtg cagcctcagt tcatctctgc tgccagccat gtaccagaat gtcagtcctt 39600

taaattaggg ttagccatta ttaaagtcta aaacctagcc tgaaggacac acacaagctt 39660

gtgtgttctt atatgtattc acacacagaa gctgcaggta aggttagtgg aactcaaact 39720

tggtctgaat ttctgactta caaagctata ctagttgttt ctgatttatg caaaataaca 39780

acagtcacca ccttggcatc tgaaaagaga caaaatcacc aatcttctgt cctgaaaaag 39840

acataccact ggattaaaat gttaccttag tatatttaca gatatccaag attatatcat 39900

atggcaaaac aaatcaacta cacaaatact caagacactc ttgtagccct ccctgattta 39960

ctgtgaaaac caatcagtat cagtataagg ccccaaacgc atcccattgt cagacaacag 40020

ggacaggttg caggggcaca ttgtttgtct ggaggaacgg aacaatgcaa aacaaactac 40080

tggagatcca gccccaagct agcagatcta agactgagca tggtgaactt ctgaggcaca 40140

gccatttgtg agctcccgcc atcttgccca gtggtggagt gcagggccag aggcctccta 40200

aactcattcc cttcctcagt gtcaggaacc acttagacat agctaatgat gctctataag 40260

cttcccatgt ccccctcatt tcttccccca gcgctgaggg aaactggtca tggtgtaagc 40320

attttctcaa gctattttcc ttcttgcctc atctgtgact accctcaggc acaattacct 40380

gggagaacct ggctgctgta ttgccctttg gaggcacatt tgacctagtc cagttaaaag 40440

gttccaccct gaagaaggcc tttgagcata gcgtgcaccg ctacggccag tccactggag 40500

agttcctgca ggtgggcggt aagtcaccca tcctgtaggg ctggcccatc caaagtgaca 40560

tggcatttcc tgctggttgg ctcagcttcc ccttcaccaa aattcctgtg gtcaaactgg 40620

tttggatttt tcccgaccca acaccaatac cttacccttt aatggctgtg gactctcaga 40680

gccactaaga ttaggttggg gtcaaagccc caaagcccca aagccctggc aataagcagt 40740

atttgccaag tctgatttgg acatcagatg ggccctctaa gcaggtcacc tgctttgagg 40800

tctagtttct aataaaatct tatttaagca ttcaattatt ttttttttaa atcctctcct 40860

ctgttctact catcgttgcc tggaaccatg ccaagattta aaggaatgct gtccaatctg 40920

tattaaccag gatgattcct ggaaggagca gcctactgca cagacatacc tgaacacctg 40980

gttctagtcc cactgtgacc tggggcaagt tctcaacccc tctgggcaca gggagaaaac 41040

ctttcctgtt tatctcatgg actgcagtga aagttgaatg gaaataaatg cttcctaaat 41100

tgccaagggc ttctgaaaca cttttgttaa taattccaca gtccagtgaa ttaatccttt 41160

aaaagcaatc ttattttacc tggaaatcat aatctggcct cattttttta accttaggtt 41220

gtggtcccgc ttgacagata gaattgcttt cctgagtttt cctctgcaga acacattgct 41280

gttgagactg aagggctaca gaaggcagtg gggccctgtc ctaatgccta tggggcaacc 41340

caaacatcct ccattttccc agaagggagt tgagggaaaa gcagttttct gatgctctag 41400

caaagcactg catagctcta tgccttgtgg aagtcaccaa atcccaaatg attaccattt 41460

aatggtcttc aaaattgtaa aatgctctct gcctctgatc tgtttctcac tttctaaggc 41520

cattgaatca atttagtaga ccaattcagc attttaaggg aaaaacagaa aaagatgtgg 41580

ttttgtccat gtagagtcac cctttgccta ttctggtcca gagctggttt tccttccagt 41640

ggttctcata tgtcctgtaa ggactcaagg ataaagggtg gctgcaatgt cacaattctc 41700

ttgacaaaat ttcactcagg tttaaaccaa aggaaaaacc acagccccat gccccaattg 41760

tagagtttgg ccaaaattcc ttaggaaaca gaaatctccc tttggatctg gtgaaaacag 41820

attcatttct tttctaggaa tccatgtggt gtatgatctt tcccgaaaac ctggagacag 41880

agtagtcaaa ttagatgttc tttgcaccaa gtgtcgagtg cccagttatg accctctcaa 41940

aatggacgag gtatataagg tgatcctccc aaacttcctg gccaatggtg gagatgggtt 42000

ccagatgata aaagatgaat tattaagaca tgactctggt aagcatgact gtctcttcct 42060

ttctctaaag aacaacaaaa ttgggccaag aaggggagct actagtggtg ccaggagggc 42120

aaagtgactc cctgtattgc tgactccctc ctgctttgct tgttggaaag cagcagcaca 42180

gcacagcaga gaggaatatg taccctgtca atccccgagc acacacatgc cttcatccaa 42240

ccccagagac aaaacatttg taaatcaaaa atgttttaaa attcttttaa ctacatctaa 42300

gcagctattt aaattgtgaa atataagtga gttttgcatt aacagaaata tcacacctct 42360

ggaattttag gatatggaaa accatcacat taaatcattt aaattttctt tcttgaagaa 42420

gtttctgtgt gaatcagtca ggtagaagtg gatggggtga gcgccgccct ttcagcacag 42480

tgggttggtg gtacaaatca taggaataac catacttgat gtcagttggt acttctggca 42540

gcattgaggg acattatatg ccagcctcac ttttagaaat tctgttgaga gtcacattat 42600

gtgcatacag accactgaca taggacacac ttccccttcc tcttcctagg gcagagacca 42660

gcatgcttac aagacacaca agcagcttct tggcatctct gggtagttcc accaggatct 42720

tcctcagtct tatcatgctg gacctgtcat tttagctact tattccaagg agcgtggagg 42780

gcaggaccag accaaaggag gcagcccttc ctgggaggaa aggtgctggc ttcttctgtc 42840

ccttccgtga cagaagatgg aagggcaggc gtaaaagccc catgagacaa agaggctgat 42900

agccatcagc tgtccaccac tcaagcctag actgaaggat gttagagaga ggagctcatg 42960

cagaaggcta aggaagccct ggacctaagc gttctcaaca ctggttatac attagaatca 43020

cttcaggagc attttaaatt aaataccaat ggctcggccg cagctcttaa gagattctaa 43080

gttaactgga ttggggctaa actctttttc agtgattcta gtgtacaccc agaggtgaaa 43140

accactgaag agagaagggt tcttaagtac tggagtgata tttttaaaat aaaattctgt 43200

tgggttccca acagaacatg ggttcccaac aaacatgggt ttgttgtagc tataggcaat 43260

tagcatttat cctctcctct ctttggtaaa ggaaatcccc caagtcccat catcaattac 43320

atgaaaataa ttattgggca ggcaaacaaa aggaagccag ttttatctct caactaattg 43380

ctcttgaccc agttccctaa tgcccttgtt ctggagtggt tcaaaggcta tgatgtcaat 43440

gacctctttc tagagtcatc acagagctga ggcaactcta cttcccccaa ttacccaata 43500

ttgaacaaca gacacatttt acatctggag catgaaaaag caatcaaatt ctttgtactt 43560

atgatcacta gcgttcttgt cttctgtgac actttacccc tgcttatgaa attgcttccc 43620

ttttataatt acaaaggact accttactgt tgattgataa ttaccttttc tgtagttaaa 43680

ataatgtata tgtttttcta ggtgaccaag atatcaacgt ggtttctaca tatatctcca 43740

aaatgaaagt aatttatcca gcagttgaag gtcggatcaa gttttccaca ggaagtcact 43800

gccatggaag cttttcttta atatttcttt cactttgggc agtgatcttt gttttatacc 43860

aatagccaaa aattctcctt gcctttaatg tgtgaaactg cattttttca agtgagattc 43920

aaatctgcct tttaggacct ggctttgtga cagcaaaaac catctttaca ggctcctaga 43980

agctgaaggt tagagcatta taaaatgaag agacagacat gattactcag ggtcagcaac 44040

ctagtgagtt agaaaaaaaa ttaacatagg gccctataag gagaaagcca actatgttaa 44100

gtttacgtgt ccaaatttta atgaaatttt actaacaatt ttaaaccata tttttcttct 44160

tcatatccat ttctaatcca tcaaacagct tatgtttaca taaaatttta tcattcacaa 44220

ggaagtttta agcacactgt ctcatttgat atccacaact tatttttggt aggaaagaga 44280

gatgtttttc ccacctgtca gatgaaaaaa ctgaagctca aaaagggttg acttgaccat 44340

acagctaatg ctgacagatc caagacctag acctaggtct tttgaactca agtccagcat 44400

tctcaactat atcaagttac tgttcagaat acttaatatc tcctctcttc ataattatca 44460

atagccccaa gctcatggat gacaaatctc tgctttattt cttgtctcta ttttttcact 44520

ttatagctcc tgttataata gcaagtttaa tggtataaac acaggatacc atcctctctt 44580

gcaacaccca tgtgcctttg atgagtcagg tagcaagctg tagtagataa tgagaaaggc 44640

cagaggctgc aaaagacagt caaaggacac gagagaaagg aaggggaaga acaggactcc 44700

aggactgttt tatattatag aaaagcaaga gctaaagagc atttacacat gttaaacaga 44760

tacttgttaa gcatagtgcc tgacacacgg cattagctgt tattttatga gattccatca 44820

gctctgcctc tgtcctcttt cttctaacat gaaggtatca tgagaagaga accttctaac 44880

ataagctgta attctaaacc tgcacttgtc cctctccagc aagaggctag cactgaattc 44940

attctactca tactacacac ccagttatgg aatgtccaga gttctcgaag aaaataaatg 45000

actttaggaa gaggtataca ttttttaagt cgctctgcct ccaaatctga acagtcactg 45060

taaatcattc ttaagcccag atatgagaac ttctgctgga aagtgggacc ctctgagtgg 45120

gtggtcagaa aatacccatg ctgatgaaat gacctatgcc caaagaacaa atacttaacg 45180

tgggagtgga accacatgag cctgctcagc tctgcataag taattcaaga aatgggaggc 45240

ttcaccttaa aaacagtgtg caaatggcag ctagaggttt tgataggaag tatgtttgtt 45300

tcttagtgtt tacaaatatt aagtactctt gatacaaaat atacttttaa acttcataac 45360

ctttttataa aagttgttgc agcaaaataa tagcctcggt tctatgcata tatggattag 45420

ctataaaaaa tgtcaataag attgtacaag gaaaattaga gaaagtcaca tttagggttt 45480

attttttaca cttggccagt aaaatagggt aaatcctatt agaatttttt aaagaacttt 45540

ttttaagttt cctaaatctg tgtgtgtatt gtgaagtggt ataagaaatg actttgaacc 45600

actttgcaat tgtagattcc caacaataaa attgaagata agctctttgg tctttgatag 45660

ttatttcttc tctcattcaa acacctgcca actaaagaca atatgactga aggaaaactc 45720

tgacagattt tcactggtat catttctctt gatcataagc cacttaccat ctgataaccc 45780

tccatagccc ctcacacccc taaaatgaga agaaaaatgc atccaatttt taagtaggag 45840

agaaaagtga gtggtatttt aaggaagact actaaatttg gtggctttga tatacagaat 45900

ctgatataaa aatggttttg aggccagt 45928

<210> 18

<211> 48382

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 18

atgtgtcccc gagccgcgcg ggcgcccgcg acgctactcc tcgccctggg cgcggtgctg 60

tggcctgcgg ctggcgcctg ggagcttacg attttgcaca ccaacgacgt gcacagccgg 120

ctggagcaga ccagcgagga ctccagcaag tgcgtcaacg ccagccgctg catgggtggc 180

gtggctcggc tcttcaccaa ggttcagcag atccgccgcg ccgaacccaa cgtgctgctg 240

ctggacgccg gcgaccagta ccagggcact atctggttca ccgtgtacaa gggcgccgag 300

gtggcgcact tcatgaacgc cctgcgctac gatgccatgg taagacccga gcccgcgccc 360

gggatagtag tcccggactg agagaggagc cgggctggaa aagcagcgga tggcagagtg 420

tggcaagcct aggtccaggg cgcggagaga tgtggggata aagtgagact ccggccagtg 480

tgccagctgg atgcatagaa gtcccttgga cgattcgtct taaacggacg ttattgcgcc 540

cccactatga gatgggagta tttagaggag caggacccag tcctgcctgc gaggggctta 600

aggtggggtg cgtagaagca ctgacagtta tccagagcta gggactgaat cggatccctc 660

acgcagctct catttactgc tagatctttc aaagaatgct ttatctcaat ctagatcttt 720

cgaaaaatgc tttatctcaa tcttattgaa agggaaaccg cgtcgaagaa gctgaataaa 780

ttaacaggca ccatccccgc aaaaagggag acgtgataag aatggacacc agatctcttt 840

gaccccggag cttgagagta attaagctca cagctttggg gtagattctc tgttcaatta 900

ccttctatgt gaccttgagc aagttattta ctctttcttg cctcagttct atcctgtata 960

aaaataagga ttataattgt acatagtgca catagtgtgg tctctatatt attaccagac 1020

acatagtgtg gtgtctatat atgttgttat taccagacac tataatatag ggttaagaaa 1080

aggaaagaaa gaaagaaaga gaaagaaaga aagaaacttc agagagagcc aaaatgtgga 1140

aagacttggg tttcaaagga tttcaataaa gtaggtttgg cggcagagat ttaggaagag 1200

tccaacttaa gagataatgg gctcttaaaa catgtgcaaa agtttagttc aaagtcccaa 1260

atgttaggtt aaaggtaatg ccttttctat tgcactaaat tctagaaagg gagcagaggt 1320

ggttgcacag taataaataa gtacaaccaa gatgtatata tggtggttaa aaaaaaaata 1380

aaaccaccac ccaaatttaa ccctgtgtaa aagccttgag taaagggatt cagcaagact 1440

gtggagaaac ttgggagaag aaaaaagagc tttggaagag gaagaactga gaaatctcca 1500

gagaaacgtc cagagaaatc tgtgaagtcc acaagtgtgg aagggaaagg gaagtgtgat 1560

taggctggga tacagaaggc taacagtaca ctttatatga gaagagctta gaggtcagga 1620

gagcacaatg ctgaggggtt tgtttggttt tattttcccc tctggaaact tggagaaagg 1680

aggacaggaa aagatgataa gagggcacca ttgtttggtt ttaaaggtcc ctccccaacc 1740

ccccacaaca agaaagctca ttttaatgag atgagagaag gggagaaagc tccctccgtt 1800

ctgatgcttt tgagcttatc agggacaccc tgcagcagaa attccagctg atccttcagc 1860

cttcagagtt gatgaggtgg ggctcacaca catctggatt tgaagaagaa tcgtgggttt 1920

caagaactct gtggtctctt aagcattggt attccacagc tacacttccc tgatactttg 1980

cactcagtaa tgacaaaata aatatttgta gaattgagtt gttgctaccc tgtgtctcca 2040

cttcctccca gaagggtaga ataaacttac atgtgtttgt tttattttgc atataaacca 2100

cgactttgta aactttactt tttgtgacac tcttttctcc acaattgtgt gaaccctttt 2160

tagtggaaaa ataagaatga agaattccta gttttcccta ataatatttc cctcttgtaa 2220

gtctagttga gctcctggtg ttaatcatgg tatcaaatgt ttatgacagg acatctaggt 2280

gaggcctgag taaactgctg taagttacag cctatctctt ctgtgtttgt tcagaatcct 2340

ttttgttgga tgaaattact ttctttcaaa ttttgctcaa gaaaattgaa ttgggttctg 2400

aggaaaaata ggctacttac gtaatgttgg cagggggtaa agattgggag cggatcacat 2460

ggcaaggtta gatggctgtg tcctcaggac ttactctgtg cagtttgggg gtgtgtcaaa 2520

catgatttga cgtgcataaa atcccagttc tgtagtgcta aagcatgtgc taaagctctg 2580

ctgtactcca gtattgcaca gatgtttcac tgtcaattgt ttatattata ctaaccctca 2640

gagaatttat tgctgttgtg gtataaacgt tcctccactg taaccactca ccgcatattg 2700

attcctagga cactggggat tcaaatcagg gtggaaaaca gttgtggagg ggcatctcca 2760

gtttcctgga gactggacct agttcaaagg agccacataa aaggactgaa gcatatggag 2820

gacaaagagg aatttcctag aatccagcta aaacacagga gtcccctcga gcaggaactt 2880

ccaaggaggt gtacactttc tagacctact ggagtaatcc tgattgctgt agaggaccca 2940

gggcaatcct ggagaccccc tggaaccact actaggcaga aaactgggaa gaggatggca 3000

aaactctcca gttagtgtag ctgtctcagt ggaaggactg ttgtcccctg aggcacgagg 3060

aagacagcat ggaagcactg acagtatgcg cttggcctgg ttactttctc tgagcttcat 3120

gttccttagc tgtaaaacag gagtgataac ccccatcttg cagaggtttt gtgaggaata 3180

catgagataa tccctgctgg tgctcactga aagtgagttt catgtgccct gggtggggag 3240

gaggggaatt gtgaggaagg gaaggtttgg gttgtgcatc agattatctg attctcacag 3300

cctggcacag tctaccacct gcctgccctc taccatctta cctaagtacc attgcctaaa 3360

ctatttttgc tatagttact cattgagcag acttatgaaa tctccccaca gcacaaaact 3420

gaaggtatcc acctccagag gtgagattct caccaagaca ggctgtgggg aaggtagctt 3480

tgagcaggtc tctagggtcc cctagaaagg tgccttttct ctgcttggct gctgcttctg 3540

aggccctggc tgctgagccc acttcctgcc cataaggctg cagccttagc cctagccaat 3600

tgctcctaaa tgggggaggc agcagccagg gtgggaacca tggcccccgc ctttaactca 3660

cccaggggac aggttagtca gatagatggt cctttgttcg ccaccctcca cctcccacat 3720

agacaacatt ctaatgtgtt tgatggactg catatagttt tgtttctaca tgcactttca 3780

aaacctttgt gtgcttatat tttataatgg catacccagt cctgttattt agccctttct 3840

gtttcttact tttgtcctga acttattgtt tttatgattt gtccatgttg ccctagatgc 3900

ccttatacct atacatcact gtgaattgtg ctctgtgtat tttacccatc tccacgtatt 3960

ttacccatct gctccctggc tgatgggttt ggagatggcg tctaacaccc tgcctccaca 4020

aataatgctg ctccagatat cttcctggat ggtctcacca gatgatttct acccaggagc 4080

agaattgctg gccacagggc atactttgag tactacaaga ctggtttcct caatgactga 4140

catcctccat tctcttccac ctactcccat ccccaaacag gacatgtaaa tttctatatt 4200

ttacctatat atcatggcag tatccagctt tattgtaggt gtaaaataca cattgctgct 4260

ttatcatgta cttttctaat aactagtgag tttggacatt ttttgatatg tttgttaact 4320

ttttaagctt cctcttctct aattgcttat tcatattctt tggtattttt taaattgggt 4380

tagctttata ttttttgact ttcaggagtt ataggttttg atctgtgatc aattttagac 4440

attgcaaata tattatgatg atattaggtc tattaatttt gtccatggta tctttcattg 4500

aacagaaatt cttcatttct tgtaattgaa ttaatctttt tttccttgtg gcttgtgctt 4560

ttgaaagttt gcctaatagt ttgctaattt aaagcttatc acctttaaca ctcctcactg 4620

gaaattcaga gatcaccacg gttctctctg ccttttaaga gataaatcta caatctgtgg 4680

ctatgcaatt cacttctcag catcaccagt tcctcatcta taaaatagaa gtttgaacca 4740

gcttacctct gaggtcccat acatatttga acaactggag ccctacagtg aaatgttttg 4800

attcagtgaa gggacaggag gatggggcaa aggtgtagag gcaggaatct acacaatgga 4860

ccaggttgga ccagagtgag gtagcaggaa tcatggcaga agaggttgtg taaactctaa 4920

aggccacctg agaagtttga gttatgttgc catgggcagc catgtggttt ctgcgtagaa 4980

gaccactgtg gtataataaa aaataaatat ttggtccttg tccccagtac ctgtcacaga 5040

gttcctaaaa ccctagaaat ttcctaagta atagaaatgt cctttgtgat tcataataaa 5100

ccccatgata atacctgggt ttatgctcct gcagacttag ggtgagatcc ctagatagct 5160

tcaagatgga accagtcacc agcaagtcca agagattagg ggatcaaaga gttggaactt 5220

tcggtaccac ccaccaaagt ccaggaaagg agggtgggac tggagattca gctctacaaa 5280

aactcttgaa caagatgtga ggagcttttt agttgctgag catttggagg ttcctggagg 5340

gtgcacaccc aggtagggca tgaaagctcc gcattgcccc catatcttgg cctatccatt 5400

cttcacttgg ctgttcatct atatcctttg tatagccttt ataataaacc aatgagtgag 5460

ttctgtaagc tgtcctagca agttaattga agccaaggag gaagtaatgg aaacccctga 5520

tttatagcca ttcagccaga agcacaacct caggcttgtg atcagcctga agtcagggca 5580

gtcttgtagg attgagccct taacctatgg gatctgacaa tgtcttgaga tagtatcaga 5640

attgaattca attataggac actcagtttg tgttttctgg agtactgttt ggtgtctggg 5700

ggaaatccca cacacatctg gtcacagaag tattctgtgt tcagtgtgac agtagaaaaa 5760

gcagtttgtg tgtatgtgtg tgtttgtgct tttgtatgct aatgagatga tctatggctg 5820

ggggccctta gatagcttcg ggaagggggc tggtcgccag aaagactaat ttatgactag 5880

agggttagaa ctttcagccc tacctctccc acttctagag ggtagatggg ctacagattg 5940

agttcggtct ccaacagcca gtgaattaat caatcatgcc ttcacgataa accttcatga 6000

agacccctaa acaaccagat tcagagagct tccaagttgg tgaacatatt gaggtgctgg 6060

gaaggtggtg cacatggagt gggcatggaa gctttgccca tccactccca tgccttgccc 6120

tgtgcatctc ttcacttgct tgttccctag tcgtatgctt tataataagc cagtaaagtg 6180

ctttccaagt tctgtgactc gttctagcca attatcaatc ccaaggatag ggtcatggga 6240

acctccaatt tataactagt tggttgggaa tacaggtggc ccaggactct cgactggtgt 6300

ctgaagtggg ggcagtcttg tgcatttgag ccttctcact tgtgggatct gatgctaagt 6360

ccaggtagag aatgtcagaa tgcatctgaa ttgttggaca ccaagttggt gtccacacag 6420

aattggggaa ttggttgatg tgaggaaaaa ccctcccagc tacataggga aatcacggtt 6480

ttggaagatg agcagaaaag ggtgggaggg agaagacaag agagaagagg gtgaggaaag 6540

aggccaggca gcagtccaag tgtgagatga taacacctga actctggtaa ttgaggagac 6600

caaatccacg ggaaccagct gcaaaaggca ttctgtctgg taggtatagg gctaagggag 6660

taaacgtggg ggtgagactg ggatcaagtc cccttcctac cacttagaat ctgtggactg 6720

ggagcaggtt atttaatctc atgaaacctc catttccaca tgtgtgaggt atgtgattgt 6780

tgtctccacc cctagggtgc tgtggggttt acaagagata gtagtgcaca taatgtgctt 6840

agcacagggc cagcccctag taagtgccgg ctgtacttgc cttctctttg caggggtggc 6900

tcaggagagg gtgtaatcaa aagatgatta tgagggttca agcaccaggc aatgggagag 6960

tcagcaggtt ttaatgcttc cacacagagg ctggattctt ctacttccgc ttaccgctga 7020

cagctgaggc ctgctacctc aagccctgaa atgcagtctg aagacttttc ctggacttac 7080

tgggcaccat tggactgtac agggctgtgg cagacaggag cagtggaaag agatttggat 7140

gggacgtcag aacacccaga ttttacttct tggctcttcc acttactagc tgtgtgaatt 7200

tggcagtggt ttccttatct ataaaatggg ggaattggag agggtggtct actgcatgtt 7260

acttctcagc acctcagctc ctccccaccc ctctctcttg cttctaatct gcctacatca 7320

cttggtttgt ctctggacaa gtctaagttt gcatgacaac tctctaattt ctccttcaca 7380

gttatttatt tattttttgg cagctatttc cttgtggaga gatagcgtag gtctttcagg 7440

tgcttccatt tggccactgt ttgcaggtga agaggtttcc aggatgctcg cgggatgtta 7500

ctctggtagt aacagaaatg atggcctggg gaatgggaag agtatgagcc cagttttgag 7560

cctgagtctt tggggctcat ttattcattc aatctgcaaa tcttcattca gcattactac 7620

atgccaggct ctctgctaag tgctgaggat agcgatcagg acctgctagg gtgaaaatta 7680

ctcttcagaa gatagcagag tggataatgg ccattcgagt tcctcttaac tacacagagg 7740

gactgggtaa gatctataaa ggagagaggt taaacagcaa agcaaaagtg actgaaattc 7800

taacacccat gcatggggat tcttgagcta aagatctggt ctgtagaaga agacacatca 7860

gaccctcata tttagcatct aagccgaaat cagcagtaag catgtttctt ctcccataaa 7920

aacttgtccc taagaagaga gacaagtgct tacaaggact cttatgttaa aggaaactgt 7980

cacaattact ttatcttcag tcatgaagcc tttcttgagg aaagaaaaat tagcaatact 8040

agtttgggaa gcttgaaaga gagattctgt ttgcaatgac acttttaaaa aggcaacaac 8100

aaccaaaaaa gtaattattc ctaaagctca ggtgtcagaa gttgtgaatg tctagggggt 8160

tgtggatccc caacctcaga gacaggaaca gcaccactcc catcctgggg ctgtggcctc 8220

aggcctctgc ttcctctttt tcagtctaaa ttccagagca gagaggtgct ggatgaccag 8280

cttcaggcag gggagggagg aagtaaaggt agtttctcag ctcatcttgt cctcgtacta 8340

ttaatagcat ctgttgtttt cttaactgaa aattggagag aaccaaacaa atcaacatca 8400

tattggcggc tttctgcaca tcactcacaa atacatctga atgctctgta ctctccagtg 8460

tttattttcc tcagccacat aatcggaatt tgaacattct tttgacactt cttttccctc 8520

acttgaataa ttagttctat atataatcca aatattagct ggagccctct agcccctcag 8580

agatatagtt gaaccttcct ggacctcaat tagttgggca gttaattctg aatgttgatg 8640

tgcctatttt gcacgtatgg agagtttgta cgtcccccct ccccaaacaa gtagaatgct 8700

gtctatttgg tacttgctca ttcatacatt ctttcaacaa ccaaacccag ttgcagctgg 8760

gttgaacccc agggtgtttc ctgctattca gtgtgatgcc acattcacct gctgtggcca 8820

acattccttg gcccaggact gcagggatgg aagccaaccc aacatcagaa gtgggcaaca 8880

atgccagaaa cctatcaacc aaaacaaatg taatttgtaa tcctccctgg atgatgttga 8940

gtgattggca actgagcccc gtcatatagt aaatcaggct gtctctcttt aaaaaagaga 9000

gacagagaga gagagagtct cccaatgttg cctaggctgg tctcaaactc ctgggctcaa 9060

gcaatcatcc cttcttgcct cccaaaatgc tgggattaca ggcatgagcc accatgccca 9120

gcctaggctg tctctttagc tcttcaatta tagggcttcc tgcaacctca gcccttaacg 9180

aatgagctag cgtgtaagtg actagctaaa taaatgtgct gcaaacctct gacaaatggc 9240

tttttgacat tattaaaatt tatttatgtg gaacaaaaaa attcttttct tctgcatcat 9300

gcatcctaca ctgcttttaa agaaactctg ctatcctttg ttttaccgtc taacatttct 9360

ttgtttccct tctgacatgt ttcagcctct tgtcatcact tggaacattt ttactgcctc 9420

atccctctag tttccatttc ctagcctagc ccagcccagt atttttgaaa cccaaatacc 9480

tggaattttg cctgatgttt aaagccaaaa ccaaatgaaa atgtgaagaa ccaaaggaga 9540

ttttattctt ctgtgactca aaatcctgtt gttattgtta aatgaaccct gtgggataca 9600

gtctggtatt tttaacacct tctgggggta tttgtcaata tttacatttg atgtggatag 9660

cttataaaag tccagtggct ttgggtaaac cctgttcact aagaataaga cccagaaggt 9720

catatgaatc aacgatcaaa gaatgaagat attctgaatc caagtttctt gttttgatta 9780

ttaaaaatac cttgcaatta gttaggcatg agagaaagct tgtgtataaa gtttggcttg 9840

taattcacat gaattctcta tacaggccaa aaactacctt ccctttggga atatacaatg 9900

accttatttg gaaattactt caggtaatta tgaagtaata atatctctat tcatatgcta 9960

aaaagaattc aaaataacat tcaattatca tctcttagaa actaaattaa acgggtactc 10020

acccaagttg aaatgcaaac aaatgcatga tatcattatg tgaccctccc ctagccctta 10080

ccctgtccat ggcagatgct aagaaaaact tgtccctaag aagagagaag tcaattgatt 10140

tagtcagcct ttcccactgt gtttggactg agcctcagaa ccctccttat ccccatactg 10200

caaggagtta ctgtcaatca actagagatg gcgctcaaga taaaattatt ttccatcctg 10260

tactaatata ataataagca tggaatctat ttattggttt tttcacaaca ataaaatata 10320

tctttagcaa gtaacaaatg gtagaggtaa tacttttaga taaagaggga agaatccttc 10380

tgacttttct ctttatcaca ttttacccac acacaatgtg gatggaatct gcaactgaat 10440

ttagcagtca cactgaaact ctgctgcatg accacactgt cctgaggcca tgtgccaact 10500

tctatgtttt tttttcccaa agctcttcca aggaagagct gccacttctc catttactct 10560

tttcacctgt aaatgaatgc tgccgttggt attgagaaaa tgggtgaatg aaagaaatca 10620

tgagaacact cctggggtga gagcaagagg aatgaaaaca cgtggtgtga gagcaggtca 10680

gagatggtgg ttaacacagg aaccaggatg gaggcctcag acttgaccct ggcctgcagg 10740

acgttggcct gcattgtctg caggaggaaa cacagaggtc agtggatatc ctgcggtctt 10800

ccattaaggc ctcttactat tctggaacat tttattttta tagagaaatc tcccacttcc 10860

tgatctcccc agttagaatt gattgatcaa agcagtccct ctccctgtta ttaaataatt 10920

aaataatatg cattagttga tccctataac cacttaaaat gtatctaaca caggagaaat 10980

ctttggtaaa tatttactga atgaatgaac aaataaataa ttggagggag ggagaaagag 11040

acagagttaa atcctgaggc tggagttttt ggtccttcag ccccttatcc cattgcccct 11100

ggcagcccca tccatgtgac cttgaattct gtccaagtct catcactttc tatgtgtgtt 11160

gtaatggaaa aaaggttaca aaacacagtc tttctggcct ggagtaccaa aatagtaaag 11220

acttctgttt tgaaagcctc tgacaaatga acacaacagc cacagtgata actttagcag 11280

gaaattgaac cacaaaaagc aagcgagcca caaagccctg aaggctaaaa atgcctcaca 11340

ggccaagagg cagttaggtt gcacctgaga cagaggctgc tttcttaact gggaaatggg 11400

tgtctccaga tatataaagc tgtgaggaga tcaccatggc ccagctctga cgagccactg 11460

gtacagcatt gttggttagt ttgcctcaac ctgcagcaga gtttttattc caagagttaa 11520

cagttagtta ggctttttcc aaaggttgag atgacacccc aactcccaca cccatttccg 11580

tcctgcccca gcttgaaacc aggcaaagtg agttaagaca ttgggagatg ggaggggaca 11640

ggaggggtag aggcctggca cccctctctc ttctgtttca agagacactt tgtggagttg 11700

tctagagaga ctgggggtaa ctgctccttc tgcccccaat ctccccttag ctccccactc 11760

ctccctatcc tgccaggcca accaccagca gagaagggcg cttagctcca gtgacgctgt 11820

gagtgaggta cttgtgagga tgcatatggt tcctgttgga aatgcacgga ggagcagtga 11880

aacaccagaa gccccatcct ctctcctgcc cgtgccctca tgcccagcca gctgaatatt 11940

cctgtggctt ccactcctgc actgtcccct cagtgtcacc tccccaccac attcccacag 12000

ccccacttgc tgtgtcacct ctcccctgag ggagaggtgt cttcctgcca gtctcttcac 12060

caccaggctc acacaactcc catccattca ctgcaggctg ccagactgag ctttccaagc 12120

acccccggac tgcctccagc cctactttct cttcaaggct tcccacttcc ctcaaaatga 12180

aagccctgac tccttggaca ccatacagac cctccctgat ctcccagcga tgctcttagc 12240

agcatcattt ttttttctgc cagattttca ggtgccctca gttccaggag caccaaatca 12300

cctggatgcc atgagctgct tatgacacag ttcaaggtgt gacatctttg aggtctctcc 12360

ataacacctc tactgcagac gtctgtctct ggtgtatcag ctgtaccttg cacacatgtc 12420

tctaaccatc tgtccactgt gtctgtctgt tcacacatca gattgaccct atgttctctg 12480

tgacaccagg tgagctctgg caccagtgcc cagcagatga ttactgaatg attaataaaa 12540

ctactcactc cacctgctgc ttttaaatgg gctccatttt ggtatgattt ggtgcgtgaa 12600

ccattaaaac ttttcagaag tcaggatttg aaacaaactt ggtaaaaagt agttaagcaa 12660

tgtcttcgtt gtaaagtgag aaaccaagtt gcttggaaac aggtgaaaag cttagccagc 12720

agcttcacaa ttcacagcct ggcactccag tagccctctc tggacaattg cccaggtttt 12780

cttgttttca tttctgtgtt ttaatttaca agttaccttt atatttctac taatacctta 12840

ctggcacttg tttgtgaatg aaacaatttt tttgagagtt tgttaattgt tctgtgtgag 12900

tgtggaaagt ttctttcata taagatcatc aattcttttt ttaaaaatag tgttttaggt 12960

tttctcctac ataaagaaag tggtgggggt gaggagctgg gggtggacca gatgctcaca 13020

ttgtatttct atgattttgc tgttggagat actgaagatc attttgctgg gctttgctct 13080

cttaaattac agaaaagtgc atttggtgaa gtttagtgtt tttagtaaac atatacaaga 13140

tgtcataggg atagaattag tgaaggaata gggttcgtga atgaatgagc tcaatcttag 13200

taatgaaatg tttggaaagc aacagaacaa gagagaaaga gcatcagaac atgatttagg 13260

taccccatac atttatacaa ataaaaaaaa gatctgattt agggtcatag aaatcacata 13320

cctaaactga tggttaaaca ataagtggca agtaacagtt tagctacaaa aaatgtacag 13380

gaacacataa ggaaagttac atgggaactt tcagagagga ccaaaaaagg tggaataaga 13440

ttatctgctc tgttatgatc ccctgtattt ctctgctcta tgttcttctc ttgggcagtc 13500

ttttcctgag atgttttcat gttattaatc aaaaataata gaaacgaata aactagatca 13560

atgcatttac aagatttttt gagtgcctgc agtatgcctc ggtgtggtgg gaacttggtg 13620

tactgaggtg agtagcatga tcctgaagag cttaggaatc actgagttct cgtaagtgca 13680

atagaagaaa aactggtaga aggaagtcaa atgaaaatac aagtttctaa agaaaataag 13740

tatttacaaa ggaatttgtc tgcattaatt tctgtggaaa agttagctat gtatagaaga 13800

aagaaacttg ttagtaattc tttcattcat tcatttattc attcaacagt gaatccctgc 13860

catggtagta ctaactactt gcactgtggt agtgcaaggg atacatttgc tgtgcaagta 13920

gttaggaatt tttttttttt tttttttttt ttttagtgct ttgtgtatgc taggctttat 13980

gcaaatatac agccaagcac agaggttaag aactttggaa tcaaaccgaa gtggttctag 14040

tcccagctcc cccttacaaa ttgtgtgatc ttcaacagtt catttaactt ctctagatct 14100

cagattttct catcagtaaa atgagtgcag taagagtacc accctcacag catgaaatca 14160

gattttctgt actctttact aactcacttt ccatgacgct ggcatgtggc aagtgctgac 14220

tcaatactgg ctcatttcct taactgcttt tattttattc attcactgcc attccagagg 14280

aaactgcggt tctgagatgt gccctgggtc actgctgagg agcggacgga gcctgggagg 14340

gagctacccc tgccttgttt cctcacactt tatggggtgg gtgtcaggaa tgaggctgga 14400

ggggaaggca aaaccagatc cctaagagcc ccctggaccc tgcttagggt gtttacttct 14460

atcataaagg ctgtgcttct gtagagacct aaggcagaga gtgccaaggt caggtttgca 14520

tttgagaaaa gtttttctgg cagcaatata aagaatagtt tagcaaaaga aatgagaata 14580

gctggaaacc cagttaggag gctgtggtag tccaggagca cactaaggca gcaacaatag 14640

gtggatggaa actggagcat ttaggtaagg gagagggatg ccagaggacg tgatggtgag 14700

ggagagggaa gagggtggga ctccaccgca ccagggcttc tggctttatt gactgggtat 14760

atggtggcat cctgggccaa gacagtgagt cacagggaag aggctttgag cctggtaagc 14820

ttgggacact tttgagacat tgggaggaaa tgacacatgg cagttgacat acagatctag 14880

acgtcaggag aaaagtcagg gctggagatt ttcggtttac agttgaaggt atgagtagga 14940

atgagatcat ttactcagtc catcccacca cattgagcac catgaataag gggctcctct 15000

ccccatgggg caagagaggt gaaaagacag tcaaagtcac tactttcagg aggattaggc 15060

tcaagttagg atggggaggt ggagaaggca tcaaacaaat aaatgaaaac tataacttca 15120

gttattgcca agtgctagga atataatgtt gctgttaaac tgcaaggtgg ctaacaagtt 15180

ggatagtctt tcttgaagaa ctgtagaaac attcattaaa aacaaaataa ttttatagtt 15240

aagcaagaaa gtgattgtac tataacaagg gagacttatt tcaccatagg aaaaaaatca 15300

taacattcta agaatagaag gaaaaaataa caattaccca agcctgcaac aacctgtaag 15360

accagatcag cttttaagag gaaagttaat tagataacag gaagaggtaa aaatgagatc 15420

atcagagatg ggggaaaagg caaatttaga aattctgaat ttaatagggg agacaagtat 15480

gtaggaatat aaacacattg tgctttctct gcacaaacac attcacatga ggatgtatac 15540

acaacccttg caggcacagg tactcagtga atgcctgcgt gccaaggagg gggacagcac 15600

tgagcaggag aggtggcagg gcctggtggc ccaggagcac tcatatagtc aggagactga 15660

tataaaagaa agatcctgct aacatgccat tacagagtat ggaatgagca aggaaggaag 15720

agggtgaggc tgtaagagag tgcacaggcc ctcatttaaa tggacgggtc accagccagt 15780

acttcatgcc tgttcctaga tgacccacaa agtgaaacga aactggatgt ggtatttagc 15840

aagtctgaga tttgctggtt taagtactaa gttagcaaac cagccttggc agccagagca 15900

ggtagggtag ggggcccaag gagatgggtg tatccaggag gccaatggag gaaggagagg 15960

aaggttttct gggaagccag ctgacatgtc atgctttgtc agtggggcct ctgcatggat 16020

tccctccact cccccccagc ccccaactca ctacggggag ctgcaaaatg acagacaatg 16080

ggtggttttg tacccagcag ccactcctca accagtgagg aatgggagag agataagaag 16140

gtgggaaaca aggattgctt gtcaccatcc ctgctcctcc tgaagccctg cgctgggagc 16200

aggtacagaa cccgctgaag gtcacctaat catttcacca tggagttctt ccccagatgt 16260

gaaactgctg ggactggagc aaagggtgcc cagagctgac tggaggagga agccagagcc 16320

acactggaac atatggagat gctagctcct gcctgctatt gaggaacttc attttcccag 16380

aagaattaga ggaccacagg agtgcttggg aagaagcaat tgtggcaacc taggtgaatg 16440

atgacctagg aggtgaaatg aggaaggagg cctttggtct ctgccaacca tacatttcag 16500

gttgcctttt gctaaagtcc tgctgatgtt aatcattctc tccagcagat cgcggcttga 16560

atcatggggt ctgaaatgtg ggtttatgag agaaaaacaa catggaaaaa ttgggcagag 16620

gctggtaaat gtgaacattc aaacctgccc ccaatctgag atggtgggac agtggacccc 16680

tggccagctc tccaggtgtt gcgtatgcac attggcaaac cacctttatt aggtggtgtg 16740

acacctgggt agcaattctc aagcaatgtg atgatctgaa tgtccctggg cctgacccag 16800

gtgaggagga cacaaaatca tgcaatatgt ctcatagagc ttagtcagtt ttgagaaata 16860

ctggactaat gttatgtttt taaagcacct aattcttttt ctcttttctg ttttatctag 16920

gcactgggaa atcatgaatt tgataatggt gtggaaggac tgatcgagcc actcctcaaa 16980

gaggccaaat ttccaattct gagtgcaaac attaaagcaa aggggccact agcatctcaa 17040

atatcaggac tttatttgcc atataaagtt cttcctgttg gtgatgaagt tgtgggaatc 17100

gttggataca cttccaaaga aacccctttt ctctcaaatc caggtatttt ctacttttat 17160

agcactcaat gcttgaaaat agatgcccta aatcacagct tggcattata tttatggact 17220

gtagataaaa gtaggaacaa ctgggtaaat agtgcactag aataacactg ataaaagcaa 17280

tctggtaaat tctatgctgt tgttccaagg tacttcttaa agagatgtcc tcttctgctc 17340

agtgaacttc tttgttgaca aaaacagtat ctttgtctaa ttcagtgctt tcaaaatggg 17400

aagaggagac aatgtatagc cagtgctaaa cagtacagtt tcatttacta gagaatgaga 17460

gcctcaaaac tcactgccct tctcctgtgg agccatgagc acagagccag agttttacct 17520

caatttacct tattgctcct tttctaggat gattattaca tgtccagcta actcaccttt 17580

ctcgcttttc attattcatc tcgttttaat gacattcttg tatctgactt tctgggaaat 17640

atatatagtg gtcttatgat atatatggat atatataagt gtatgtatat acatttatag 17700

acatatatac acatatgtat atataatata tataatctta agacacacac atacatatat 17760

gacacatata catacataca tataagacac acacacatat ataatcttaa gacccttcaa 17820

gaaactattt tgaggcactt ttataagaaa actttttgga atattttggc aatgagaaca 17880

aaaatgacat tttctttaaa ttatccaaat taaatgtgtt tccatatttg aagaaaaaaa 17940

gggaaaatag gagccagggt tatctagaaa catgagtttt cattaagaac taactgataa 18000

aggacatata attgacaatt caaaaatata tatttactgt tgttgttgtt ataaaataaa 18060

taagtccagg ccattggaaa caacaacaga aaagtcagct gtgtttgctg agagcagcaa 18120

gcgggtcctg cagctggcat agaggcagag ctggaggcag ggtggagcgg ctggggctgg 18180

agtggtcagt gagaagagtg agagaacttg ctaaagaacc ccccaacccc accctggggt 18240

caggtttact tgctgaagaa cctccttggt ccccttgaga gctgtgtaaa acatccctgg 18300

ggcacaggac attccagaag tgaggtcaga gtcacaagct aggatgcagg ctccatgagg 18360

acagaaacct ctgctgtctc actattgccc aatgactgtg cacagtgcct ggcatatgat 18420

aagtcctcag gaagcattgc tgggatgaat acatgagagc aggaaggtat aggggccaca 18480

gtcacagaac agcagatagt tccatttcct ggagcatggg ttctgactag gaaaagctag 18540

aaagttggat cagggctaga ttgtagggct gagaagttca ggtgtgattt ggaggaaagc 18600

attgtctctt ctttaaaagg cacccataag gaggtaggag gtgcaggtct tggtggaaag 18660

gcacagcagc tccagaaatc aaactgcccc agctgggact ctgaccagcc ctttaccagg 18720

ctgtaacaga gaggggatca aaagtgacac tgctgtttcc agcagctgct ggaagagagg 18780

gctgcatttt gcagggccca gtgtgggggg aaagggatgg ggtggaatag aatgtttcca 18840

ggagcactgc ttcttccaag actacaggcc cagcggcctg gtgtgggttc ttcctacagt 18900

tctttgaccc taatcaagcc cagttctccc agtcctagtg gcctgggcac ccacagggtt 18960

ataacctgta tttctagcac atttaatcta atttatttta gctagatcat taaatacctg 19020

aaatacaact ctcctcatga catcttgaaa tataagccat gctttgtata tatgggatgc 19080

ctggtcctgt gaggtttctg atataataga ggaatacatt cctttgtcct cagaggcatt 19140

tctctgaatt ttttttcctt cttggcagcc ataccttaag caaaagatgg tagcatactc 19200

tcgacattta gtcattcccc tgggtacctg tctgtgtgcc aggcactgtt ttaggcacca 19260

gggatactga ccaaaaaaaa aaaagttgcc cactggagct tacatattca gtgggggaat 19320

caataaatga gtaaatggtt tggtatgtca gaaggtgcta aggctgtgga gaagaattaa 19380

gcaggagggg aatggcaagt gcagcagggg ggaaggtata ccttgttgtt tcaaagagat 19440

gctcaaggaa aggctcacat agaaggcaac atagcactga gggtcaacgg ggagccctgg 19500

gcttcttggg gcaaccaaca ggaagcagat tgtccttgat gttctcataa tagacaacag 19560

tggcaaaact acaactcttg atttttcaac attgcctatg agttacagac agccttgcct 19620

gtccaacaca tctgtgagct gctaattatt aggttgtcat tctggggcct ccctggctat 19680

gatgtcagcc atgaattatc ttccatggta atttgcactt gtggagttgg ctaatgttaa 19740

ggattcagga gttcccctgc tgaagcacac agacattcct aaacctcaca gtggaacaag 19800

ggatgcattt ctaataatct tgttactctt tggattagta cttttcaaac tggagtttgc 19860

agactcctgc attttgaaag cttttaatga aatgtcattc tcccaccctc cttcccttgt 19920

ttagtgttag ttcttagaaa tcagtgtaag aacgtctgga tctactggag gactccctag 19980

tcacagagta ccagacatga cttcagggca gcatgtctgt atttgcagtc atcttcatgc 20040

cagtgatagc cataaaacct caaagcttcc tcaaatgtgg ctaacagagc cagttggagg 20100

aaccagttgc cttgctgcac acagagaacc tgggttgcac agtgagagcc atgcttgact 20160

actgataaca aaaggctatc gtttgttacc actatgtgca gtaataagca cccagggaag 20220

aatgtacaca atcccagtgc cagcaacatt tagatgaggc cctcatgatt aacacagagc 20280

tcaataatgc tctaatgctg gcattattta tatatttata tattttttga ggcagagtct 20340

tgctcttgtt gcccaggctg gagtgcagtg gtatggtctc ggctcattgc agcctccgcc 20400

tggcaggttc aagacattct cctgcctcag ccttccaagt acctggcatg tgcctccacg 20460

cctgaataat ttttgtattt ttagtagaga cagggtttcg ccattttgcc tgggctggtc 20520

tcaaactcct ggcatcaagt gatctgcctg ctgcagcctc ccaaagtact gagattacaa 20580

gcgtgagcca ctgtgcctgg cctgctggca ttaaagagaa catgacaaag ttatcaactg 20640

ccatattaaa ataggcattc aaggactact tgctgacagt tttaagtttg aacactgatt 20700

atcccaagtt tccaacgaac gtcttggagt tgctaatact cttttgtgta tcatgcttac 20760

gtgcttaggc attttctgca ttggtattgt ggaagttgag atgtacaaat atataaaaca 20820

atatctgaag cctcctcaca gtttgcaacc aagttcaagc aaaacaatat ccacaacctt 20880

aaactaatac aacataaagt gaatgtaatg aggatattga tttgaatttt tatttacttc 20940

ttaggggttt tgtatttaat gaataaatat gttttggttt tactgactct tgagctttag 21000

taaattaagg tgtttaacct ttgcatgtta atatgtatat taagtaatat aataaaaatt 21060

gttaataaat atccatttta tttattttgt tccttaggga caaatttagt gtttgaagat 21120

gaaatcactg cattacaacc tgaagtagat aagttaaaaa ctctaaatgt gaacaaaatt 21180

attgcactgg gacattcggg ttttgaaatg gataaactca tcgctcagaa agtgaggggt 21240

gtggacgtcg tggtgggagg acactccaac acatttcttt acacaggtaa ttgtttcaaa 21300

aggattgcat gggccaggat gtccagataa gcactgtgtc tcttttgcct ttgtaactgt 21360

tattactctt tttactgcta tttaatatgt aatgtatatt atatgatcta taatatatat 21420

gtaatataca ttaaatggga acatgtgcaa atcttacaaa cattactaag aatttattga 21480

tttcatgatt ataagtcaat tccatatatt tatgtatata ataaatatat aagacataaa 21540

atatgtataa aacatatata taccatacat atgaaaaaca aaaagactat tgagccccaa 21600

ccaagtcatt tatgatagat tcttatcaaa aagcaacatg tcatataaaa actttaagca 21660

taaacaaaat aatgtaaata aggaccttta taactcaagg tggatttgat ttgtaggttg 21720

cacaggtagc agaggctgga cctcgagtgt gagtcctaac tctgctgcag actaggaata 21780

taagtatcct ggaagccaca tgaccctggg tcctcaggag agcatgcact cttcctaccc 21840

actacctgac tacccccacc actctctttt tttctctctc aaattggtgt taagtagttt 21900

tcatcagaat aactctttta tgatttttcc actttaaacc tataatgtcc caaaactttt 21960

gaatctggtt ccacctttcc cacttccccc accttgcact gcatagatct ttgaagtatt 22020

cccaaggaaa ggaagaagag gctggtaaac agttcttcta tcagcattcg caaagaaggc 22080

tttggcgtgg gaacagccct ccccttgaag aggctcctgc ttctaccatc cttatgtggg 22140

ctgtcaaaga ccatgctatt ttgataaaag cgaaactcaa aataccccaa acaataagta 22200

ggaaataccc aaaaacatta aaagggatta tcactaagaa agtaattttt attttctttc 22260

tagtactttg gagtattttc aaataatgaa tgcatgttac ttttgcaatc ggaaagcaaa 22320

atataaatta taaaaaggaa agaaacacaa tattgggtca aaagtcccca ttcctaattc 22380

acttgaatga ctttgagcag gtcacttagc acctctgaat ctgctagctt agcttcaaaa 22440

taggggatga tcatgctcaa agttccacgt tactgtgaaa attatatgag atgtaaaaat 22500

atgtgaaagc agtttataaa ctataaatag ctatattctg gtatgcagcg tcatggaggt 22560

tgcctgctta taaaaatatg tcagtcaaaa tcataactca caaactaaag ttggccttca 22620

cttaagaaaa taccagtatg aatgtgtgct aattaaattc cttcacagat attttacttc 22680

ttggtaaaca caggctaagg aataatttca aactgagtta aactgacagg ttgtgaaatc 22740

aatttatggg ttatgaccag tagtagggct tttttttttg acagataaga ctaaaataga 22800

ttaggcactt acattactca taagaatgat aaatattgtt ccatgggaaa agatgtatat 22860

aagatgtcaa gaacagcccc aggcacatag cagacaatcc ataaatgtga tgaggatgac 22920

gactagcact ggacattgtt agcatggata acattgcaat caaagagttt caacaactcc 22980

atgagggtta aatgaactaa gagagccaag acgaggaaat aattggagca ggtcactggg 23040

aagcactggg taggggtgaa gactcaacaa agtccttctg actctaccac taatggtggg 23100

gtcctttcaa tgcctgatta ccctcatctg tggaatgaag gtgataacaa cagtacctat 23160

tataaggggt cactgtgatt taatatgtgt aatgaatgta taggtaaatg gccttagcac 23220

agggcctagt atgttagctg ttcttatttt agttatgtgc tggctcactg gataagacct 23280

gtgaaaaccc aaaccttagc ttttcactga ggcagtgtca gcacccttgg acacacagtg 23340

ggagctagat atgtctcctc ccccttcttc ctttagaata atggtttcaa aaagtttaaa 23400

ctatgacccg tagtaagaag aacattttag acagggagcc aaaacacaca aacatatatg 23460

tgtgtgtgcg ttcacatacg aacatgcaaa agcacttgta tgtgcaaaca catacaagtg 23520

aaagaaaact ttcacaaagc acatggaatg caccccaact tttctgtttt actgttttat 23580

tcacttaatg ccagctacca ctcattgaat tgattttctt gtactgtagt taatccatgt 23640

ttctaaaaac tcagctttaa atgggacgtg accaaaataa tgcatgaaca ctcaaaggag 23700

tttcatccct tcattcagat gtgaacattc ctgtgttccc cttgacttgt ctttacacca 23760

tcttaatcaa tgctgtctta cagcaacaat tagtcattca tagcctgtgg tttgtgtcca 23820

tcagtgagtg tgccctgata gacctgaaaa gattgggaga ccatgcccag cttcccatta 23880

tttctgtagc ctgtcctttg ttcccataac agtacagtca tgccttagta tctgtggggg 23940

attggttcta gaactccctg tggataccaa aatccatgga tgctcaagtc ccttatataa 24000

atggtgtagc atttgcatat gacctgtgca catcctcgca tatattttaa gtcatctcta 24060

gattacttat aatacttgat acaatgtaaa tgatatgtaa atagttgtta tactgtattg 24120

tttagggagc aaagacaaga aaaaatatat gttcagtaca gataaaacca tcctttttta 24180

gaaaataact ttttgttcat ggctggctga atccaacaat gcaaaaccca cagttagaga 24240

cagccaactg tatgtgcctt tgctaagtag ataaatttag ttaatgatga aggactggat 24300

gaaagaaaga tttgaggata aaaattagag tttgcttatc tgtaaatcct gaagtaaata 24360

attacagtct ctgagaaaac tgagataact aatacttagt catttacttc acatttcatg 24420

attgcttacc aagattaaac ttgttctgat gaaaatcttc acttttcata ctcatcgttt 24480

tgatgagtga acttaacaat taaaggtggc tgggcacagc agctcatgcc tataatccca 24540

gaattttggg aggctgaggc agaagaattg cttgagccca ggagttcgag accggcctgg 24600

gcaagatagg gagaccccca tatctacaaa aaaaattttt aaaaattagc caagtgtagg 24660

ggcatatgcc tgtagtccca cctactcagg aggctgaggt gggagaattg cttgaatcgg 24720

ggaggttgag gatgcagtga gctatgatca ctccactgaa ctccagtctg agtgacagag 24780

tgaaaaccct gtctcaacaa cagcaacaac agcaaaaaca ttaaaggtaa ttccaggaca 24840

caatagggat gaggcatatt gcctgtttaa ataaactttc ctggccttaa gaaaacaact 24900

ctgctgaata attttatgga attagaatga cattaaaata atgaatctga taaatatgat 24960

tagttaataa aaatacagtt aaaataatac atcccttgga attttaaatc ttactgagaa 25020

ccatttctgc acgcacagta tccaatatac aaacacacat ggaagaaaac ctttccaaag 25080

ctcgtggatt gcaccctcat attttctgtt ctgttatttg agtttttaaa tgctggttac 25140

cacccattat attaatttta ttgtattgta gcttgctcat atttttttaa aactcagctt 25200

caaataaggc atgactaaaa cagtgcatga atggatccct tccaaatgtt gaaagttatt 25260

ccagtttctt tcctttccaa gtaccatgca tatgcagcta accctagcat tggctgacca 25320

gtctaacatc agagtgaaga gaagggaact tccctctaca aggcttacaa aatgataaag 25380

ttacaaagat aagagctaaa atgcaaacgc tttctctaaa gaagttaaat cctagagtca 25440

cgaactggtc catgttgctt cctttagaga aaaaagtagt aaaatcagtg caggttgaaa 25500

cctgcagcag gtacaacagg ggactctatt ctctgtgctg tccttgaagg gactccaaag 25560

atgctgtttc tctgtttgcc tcccagtatt ttaagtgagt tttggattct gcaaatcaat 25620

ctggctagct atggccttgg gtgaatataa gcaccagttt ctatagccgc aagttaaagg 25680

gatttgaatt gttaattctg tcatccctcc cattctaaca tcctgttacc tagaaaatca 25740

tccatacata gagcctgaaa gcttactttt caggatttgg ggaaataaaa aaaaatttgg 25800

tagaatacct catactttca ttaggctact gatatgaact taattattcc tctaactcct 25860

ttgaggctca gaccttctgt cccctgtctt atcactctca tctaatcacc tccaagcaat 25920

gtcccacgtc actgagaatt tagtatccaa tttcctatct tctgctccac tccagctcct 25980

gccttcacct ggcagagtgt ccatgtgggt agttgatcca ataccccaag ttcagagtcc 26040

cttgatctcc tcaatccagt gactgacacc tccactccat ttcagcaact tcacttctgg 26100

ggccacaggc tgagacttga ttatctccag aatttctcca tgtccaaaac tgaaatggga 26160

tagttccctt gaccccttcg tgggactcat gaaggggttg tcttgtttac tcatcctgca 26220

gctctcaact cctcatggga gggagagcac ataggtgagt gggtgcagag ggtgggacaa 26280

gtgcttctgg gtgctggcag gagtagaact ctgtgcggcc ccgcagcagc atctaggtga 26340

gtatctgtga cccctggagc cgcagagggc atgtgttaca gtgtgctctt ttagctttgc 26400

tgtccatgga cagcttaagt gttttaacag ctcagtgtga cagccctctg tatcctgaac 26460

tcttgttcag caaccaggaa ggatcagctt gcacgaacaa attgaagatg gtgaatgtgg 26520

agaactttat tgctgatgaa aatggctctc agtgggatgg ggagctggaa aggggatgga 26580

gtgggaaggt gtctccccct ggagttaggc catccccagc tgaactcttc tccgaggtcc 26640

taccgtcaag ccatccctct gaagtcaagc tgcttttcac cagtgtcaag cttcttctct 26700

tctctccttc tctgccattc cactgccagt ggatcctggg gtttttatgg ggacaggatg 26760

gggggcaggg cgggccaggg tgattttgga aaaggcaaca ttcaggcagc aaaatgggga 26820

tataaagttc tcactttggg ccacgggtcc aggcttgagg gtggagccat tgccagggac 26880

cccacccttt tctacctagt atttccctgc ctccactctc caactctctt atcacaggcc 26940

tcctatccat aaatctttct cactccctta ttttttttag agctcacctt cagacttatg 27000

aaagctcaag tcttttgaat tcctctttcc ctcccagtcc agcaggcttc acttctctcc 27060

ttgctagcct ggaagtgtcc attacaccct tgactgtgtc gcggctgaca catggaaggc 27120

aggtgtctgg tattgttaaa tgaacgttaa caagtgcaac ttctttaaaa agtaagattt 27180

ctttatttga ttgatcagtt ttttttttaa ttaagtatct aatttccatt gcttctggat 27240

ttaggcacac atattgaata taaaattcaa aacttttcat gtatctaaag aaataatttt 27300

aatgaccaca gaattttgct cttaaaacaa atggagcttg caatataatt ataactaaga 27360

cagactacaa gcacctgctg tgttttcttc atgctgccat caggctggtg catagtctga 27420

tgggatggat gcatccctgc tgtaatagga ataagtaaag agggtgggag actgttgttg 27480

gtgtttacaa tcattattca tctctccatt aattttcatc cttgcagttg agataatctt 27540

gccttttaga acgagaaaac aatcttatta cttacaacat tcacaagaca gggaaacact 27600

cctcaacaaa gaaccatttc ctttgggcat ttgataagaa ctgctgacat ctgtgtttat 27660

taagtgatgt gggagaatca gccaaggggc agaggtgtgg gtggtagctt caaaggtaat 27720

ttcctgactg agtgtggttg ggcatgttgg caatcccagt gtttgcaatt ccagtgcttt 27780

gggaggctga ggcaggagca tcacttgagc ccaggagttt gaggctgcag tgagctatga 27840

ttacgccact gcactccagc cagtgtgaca gagagagacc ttgtctcaaa aaaaaaaaaa 27900

aaaaggattt tgctttcctg gtctgtttga atgtggtaag cacaagaact tatttgtact 27960

ggtggtcacc cacttgggtc ctttcagtaa acttctggcc atggcccagc ctgagcacct 28020

gcttccttta cagagcaaag ccagagcctt ctggtttgta atttttgcag ccacataaca 28080

ctgatccatt ttagctgtct catgaaactg ctttccaaga tgaagtgaga ctatccgaaa 28140

ccaaaatcta aaaagaatga ctacagaacg aatggcagtc caaggtcatt tggcagggta 28200

gaattataag gaaaggtgat ggccagctac tgtgaagtct taagcccact cataggtacg 28260

aatctcttac caggactgaa gtttgtgttt ccttaacttt gctgcttgat ggtcttctgc 28320

aaaggatctc atttctccag ctttcactgg gaagtggcat ggggttttcg aaagtgcttt 28380

aaactctaag aactggtttc ctctcaggct tagccaccac tagctgtctg actttggaca 28440

agttactcaa ccactctgag catcagcttt ctttttttaa aatggtgaca gtgatatggc 28500

taccttagag agttgtggag attaagtgag ataatatatg aacaaaatat ataaatatgt 28560

ctatattaga catatatgat ctgttaaata tatttatata tatatataga aagaatatta 28620

gagaactgac atatttatct tagacctact atgttctagg gcccagttct gctcatgtat 28680

tatctctcta tatatatagt attgagagag agaaagagag agagagctag agaaatagaa 28740

accagtagga tatatcctat ctgaagaact ctaatacagg tatttcactg aacaaaatgg 28800

cttctcctta aaatgtattc caacaccaat ttcccctcct tgcttctcat acatattaat 28860

tcacagaaat tatattggtc ataaggcggt aggactatga ataatttttt tcttctgcat 28920

ttgccaaata ttctgtaatg ctagttttat ttttataaca aatttttaaa gtccctgtaa 28980

accctataaa attaactact aatcctaaac ttgaactgta atacatattt ttgttaaata 29040

aaaaaaaaga tatcctaaat ttgagaatct aagctaaatt cacaaaatga ctaaatgtat 29100

cttcttactc ttaaacgata agaaaccagg aatgcagtgg tttaacccaa acagcctagg 29160

tattcttcac tcatttctct tgtcctcagt atatgtttcc cccaggccaa gaaacattta 29220

tggctaggaa ttcatgactg ctcatctgta agacactctc caagtactct gtcatgattt 29280

ggaaccagag ctttggcctt ccattccaga tgacctccag gttctaagct ttatctgaag 29340

tactccacac tacattgcac accagcatga tgggaaggaa tgactcaagg gaactgggca 29400

ttcagaccac tcaccaacta cacacagcct ttcacacagc agtagacaac ctttgtagga 29460

agacctctgg ccaaatattt atatatcagg agtatatcaa tgtatggtca cagttacctc 29520

ctttttaatc tcaagaaata aagtcattac ttctgaaaag aacaccagta atggaactac 29580

acgggcatgc caagatgaaa attagtctct ctctctctct ccaccccatt cttttggcca 29640

ctgcatttta aacaggccat agtgatagaa ccagaaggta gctgtgtttc gattattttt 29700

gttttcttat ttttaaacat tgctttatag aagcctgttc tataatgtaa gaagtgaata 29760

ggaatcctag tatctggaat gatgctgagc catggagatg gccagattcc atgtgaagag 29820

aagaggctgg cagaaagcta agtatttgct cgctttccct accctgtatt catggagcag 29880

ccattacaaa agatgacgca ccagaccatg ggcctggtct tgctaggtgg tggagatcca 29940

aaatgctggg cactcactgg agctccgtga agatctgttg aatgagtgac taaatgtgtg 30000

gtgaactgtc cctacctaca gggttccctg ctttgcccag cctatcaggc ttgggtacaa 30060

gcagatagcc tggattggat ttcatgaggg ctttgtgccc actctgtata ctaccggaag 30120

atgcataatc tctttagact tcctgggtca tcactgaaag gccagaggca agaatttgag 30180

cagtaaattg aagcatgtgc agagtgacat agtctgggca tagataacag tacctgaccc 30240

ctcaaattcc ctaaagttga catgacagct tgtgcatcat ggtttggtac agagatgact 30300

gaagagacag tggttgctcc tctggttgct ggacaccttt tgaatgggct gtgatgactg 30360

gagagcgtgc tctgtcctgg tcacctgaag gtgctcctag actagcagaa gcagagacag 30420

cagccaggcc tgactggaga gctggaggga ctgggtgcag gggacttcta ggccagcagt 30480

ttctgtgctg tgctcggagg ggctctggtg gcaggaacct agtcctgggt tccagctcta 30540

ccagggtgca gcctgaggaa ctcagctttc ctatttattg gcttgccttg ccacaaagga 30600

tttcttgtgt gttaaagata ctaatagctt aaacaaacaa acaaacaaac aaacaaacta 30660

ttgatttaaa ccactaatca aatctttggt tttacaaatt tagaaaataa aattaggccc 30720

cagggactaa atgtctctcc tatggtccag aaaccagact gaaactgacc tagtagcaaa 30780

acccattctc ctgtatccca actcccagtt caggttcctc ttcattagct cccacccagc 30840

tgcatagttg aggacactga attgagggga cctggggtaa agggctggca ctcaggcatc 30900

gctggcttct acatgattcc accacatgga aagcatcata atcatgtatg atccctagaa 30960

aagtgtttgc taaactcctt aagttgtcat tcaacaaaca tttatctagc acctaccaca 31020

tgctaggcaa attctagaga cacagctgcc cctgccctca tggaactaaa tgtcttgaaa 31080

gagatactgg ccctgatcag attttttaaa aatacataat tatgcaaata aatcatatta 31140

tatataatta caaagcatgg tatgaaagga aaagtacaag atgctgtgaa agcgtataaa 31200

aggaggactt ggccaagtct agaggatcat cagtgagtgt tccctgatgg actaacatta 31260

gggccaagat cagaaagatg tccaggagtt aacatggtca ataatcactg ttaatcttta 31320

ttaagcattt cccttgagtc gagtcaggct tcacttgctg cattaacttc tttaatctcc 31380

acaataactc tgtaaggtag gtgcttttgc tttttttcca ctttacagat gaataaacaa 31440

gaagtaaaaa gatgtaacta attcacatgt agccctctgg cttcagaagc tctttactcc 31500

ataaggttcc tgggcacgtg ggtgtggtct aggaatggga agaggctaag aatgctgaag 31560

ggcagactga atgttccaga tgaagctggc gtgataagaa agggccacat catgtgggtc 31620

ttataggtta tgatattcat gttaaaactg ttaatgtttt tgaaatatct aaaaccagga 31680

agtgcatgat catatttatg ttttagaaag atcactcagg ctgctgtgtg gagactggag 31740

tagaaagaag catagaagag acctacagag actaggaagc tgtcacagta acctaggcac 31800

aagatagtag tggtcaagac tagcttgctt gaggaagtgt agagaagtag acagataaaa 31860

ggagcatcca aggagtgtac acttctaagt aatttaaaag aaagagtcaa atcccttaga 31920

agcaatcatt tgaaataacc taattaccaa tcctttttat ccagtcctct tatgttgtcc 31980

caattcagga attcttaaac cagttcttgt gttaaaatca tagcatggac tgccttacaa 32040

cattctatac ttcagtccat tgaaaaccac atcaggccag gtgcgtggct catgcctata 32100

agcccagcac ttttggaggc caaggcgaga ggactgcttg agcccaggaa ttggagacca 32160

ctgaactcta gcctgggcaa cagaatgaga ccctgttgaa agagagaaaa ggaagaaaga 32220

aagaaaaaag aggaaggaag gagggaaaaa aggaaaaaag ggagggaggg aaggtggaag 32280

aaagaaaaaa aaagaaagag aaagaaaagg ctgcttcctg gcacttctat taccatgtga 32340

cacgagaagg gcaattgtcc agacccacac ttaggttcac cacagtcctt ggttcccagg 32400

acagagtcct ttgaggttcc ttcaagcacc cacaccatgc tggaaaatgc aggctttata 32460

aactccagat gtcagtccac aactccacag ccccattggg tttggttcct ggatgaattc 32520

tcggctagat gactggaata gagttctggt gaacatcttc cttcctgcca agatgacaag 32580

tcaggtttct aaagacagat gtgtagctct ttgtggaaaa aatattctgg gcacgaacat 32640

cttcggctga acggcatgga atggtgtgga aaatgtgaag ccctggactg gtttctgtgt 32700

cagtggtttc ctgggactgc cgagttgaag gggagcagaa agggctcagc tgtggaggag 32760

gcagagatta gccaaactag accagctatc gccccagctg catggatagc atcttccttc 32820

ttgttcctga ttaagctcat ttaatatgcc aatgacaaaa ggatcatgaa ggtctttcca 32880

aatgaatgaa tgttttctgg agctgagatt gctggctcct atttgcatag tagtgcatac 32940

tgaggcatct gaccggatta ctggattcct aacttttaca ttctttttaa gctcagatat 33000

ttctacggaa ggcagaaaga taaaggccaa aaaaagatca ggcttgccta gctcttgggt 33060

ttccaggaaa ctgacagact cctgatatct gcaactagtg ctttctctcc ctgggcgggg 33120

cgttctccgc caccttgtgg acatttgggg aactgcaccg ccccagcatc tatgatcaga 33180

ggaactgaca gaggctgcca taactacctg gtcccctgaa ataaaaacca ggcaccaggt 33240

ttgcccaaag gaggaccaac acatagcaga atgaggctga gtaggatgtt aaccttccat 33300

gctggtcttc cattatcacg ttatctgcca ctgtcacaga caacatctgc atccgctaca 33360

gatggagatg gaacaaaggc tttcactgga tctcaagttg gatgcttctg taaaagtaaa 33420

gttccaagac ctttttcttt tttttcccca gtgacagcaa gaaaattgga tgttactatt 33480

ggcaacaact atttcctgct tgtgaagcgt tggtatttgc agtgcttctg ccctgtggtt 33540

attgagctga aaggggggct gctcttgaca taggagcatg gcaggattgg attcttcata 33600

acatgctacc cagcttgtct tatgtgaaaa gagaagccca gaaagagctt gggacacaaa 33660

gatgcaaatc ctgggtttgc atttgatttg ttaagtgtcc tttcacaacc agcttgcaga 33720

ctttagggga aaaaaaatac atggtaggct tgtgggtagt ggagaaaaca taagctccag 33780

tacctcacag cctgggtttt cagtctgcct cctcccaacc ctcatcacat accagctgtg 33840

tgaccgtggg tactctgtct atgacaatca attttcttgt ctgcagagta cagataatag 33900

aggacctgcc ccatgggaag cttggaggac taaatgagat taagcccagg gcctggctca 33960

tggccgattc tcgtaacagg tccatgtgtg taatcaacat gtgctgaagt tctctggacc 34020

ctctgtaaat atctaatagg aacgactagc catgggtgtg atgagggaga tgctagaaac 34080

agtaatgggg tcaaatgaat ctgtcccata gtctctaatt tgaaacagct gaactgtgcc 34140

attgtctcct tggaggtaag atataagcca cagagtgcct gaagcaggtc tacagttaag 34200

gaacttaaat gtgctaagtg attaaaggct gggctgccta ctccctgtat ggcgacactc 34260

tccaccctcc tgcatactct ccactactcc aggcagtggt gggcctccta gcctctgggt 34320

gatgagcagc tcccacatct cccctttgag acagaggcct gcagaacctg accagtgtga 34380

gggctggctg gggtcaggag tgtgcctgct ggttggccct caaggtgaat gtggctcttc 34440

taaaagacac ctgaagggat gcagaattta gcagcaaacg ccaagaacag ctctttgtca 34500

tctctgcatc ttctactaaa gattttaagc tccgaagtat atgtctgggg tttctagaga 34560

ttgctattcc cttataaggg aaaaaataca cccttattgt atatttattg agcacctaca 34620

atgtgccagg caccccagct agggagttta gatatttaat cctctcagct gccctgtaaa 34680

atagggattg tttaaaatgg atttcgccca ccttataggc aaggaaactg aactacagta 34740

tggtaaagtg gcttggctaa tccacctggc tagtgagaat ggagccacag tgaacctgga 34800

ggatactctt cacggcgttc catctttgtg tgccccacag cacaagcaca atgcccgggt 34860

cttaggaaga gcttggtaaa tgactgctga gttgagaatg gtaggattca tgtaccaaaa 34920

gggcatttgg aaatgggcaa atgcccagat agggctctga aagactagct atgtagagca 34980

acagatggca aatcatcaat ttaaaactct gtcatccagc cagtagcccg ctggcctaca 35040

gccagccatg tatgtacaag ggctgacttg atcagctgtg gctcttttat ttcaggcaat 35100

ccaccttcca aagaggtgcc tgctgggaag tacccattca tagtcacttc tgatgatggg 35160

cggaaggttc ctgtagtcca ggcctatgct tttggcaaat acctaggcta tctgaagatc 35220

gagtttgatg aaagaggaaa cgtcatctct tcccatggaa atcccattct tctaaacagc 35280

agcattcctg aaggtaagtg aagttcaggg gaatgttcca ccaatctaaa atttagatgg 35340

ctggatattt tgctccttcc catttttttc ctttaatgtt tcaggaaaag actataatac 35400

tgttgaagaa tttagtttct tccttgagtt tgccctcttc atggatagat ttaaaacaca 35460

cccaaatgcc aaaagcccca cttgcttctt caagtcttct cagcatcaga ttccatacat 35520

cactgcttcc tctgcctttt aaaatcttga tcctgacatg ccaaagccac tctcagcttg 35580

tgtttacagg agagataact gaactaaagg ggttctaatt gggcagagcg gggctcaagt 35640

cctggccccc caactagagg cagtgtgcct taggcaagtt gcccaatctc atgggcctca 35700

gtttctttat caaatgaggg agtaccttat ttatatgtct gagtctattg cgggatctgg 35760

ccagcagccc gcaatgcaat ggggctctct ctttgttcct aggcagattg gcaggttgag 35820

aaataataga cacacacaag atagtgaaag ctgggtccag gggggtcact gccttctggt 35880

cctgcggtgc caacaatgca ctggatatac cagcatttat tattaagttt agtgagggca 35940

ggggtacgtt agtgagggat ttagggtcat ttgattatga ggtgagatgg tcacatgggg 36000

atgaagtaat tctttaacat aacatttgta tgtagaagta cagtacattt gtatgtagaa 36060

gtacagtatg agataagaat ttacaatata gtgtgtgcgt cagtaatttc taacagagcc 36120

ttagaacaga aacacagtct ttccataacc tatgattagc aagatattaa tcagcagtaa 36180

caattgcaac aaaaactggt tacaaataat ccatggaaac aggacatgaa gctagataac 36240

cagttagacc agaaattctc agaggggagt acgccttaac cctaaagagg cctagaagag 36300

cggcggcaag atgagcgtgt ttatagccct atcttatcca tatggacagg ccccccccat 36360

gcgtccgttt ataggctccc cacaagggtc acattccatt cccagagcta tgaacatctg 36420

cttttctggg ataggaatct tggtgatggg aaacctccct gactgcacgt ccatttgtag 36480

gctctctgca gggggaagca catcacgtgc tgttggctca ttgtggcagt ccaacctggc 36540

attgtcttta cacaatcctg catgcaattt tgtatttaca ataatcagga gcatttcatc 36600

ttttattccg tagcaatagt ttcagggcgt ctccctacat gagtctcaga gtaaaaatac 36660

atttttatat cattgatttt catatgtgta cagaacacac tcctgtgctt gtctctcact 36720

ttcccttttc tgttgactat gatcatgtca ggaagatgag gtgcacagca cccaggtgag 36780

cctgtagtca caacacagcc ctctcagtag ccagatggga aaggggccca ggaccacact 36840

gacagcgata gcccagctgt acctcagcag gccttttata actgcccctg ggcctttcag 36900

agctcctacc gccaccacta atgctgtccc aggtgggcca atgggtagct catacagatg 36960

tgatgagctg gtagatgttc agaatattct taaattaaga tgcattccag gtcttattac 37020

caggaaacaa aatagaagtt cctaaggaaa tttcatggtc aaaaccaaaa tgcacacaaa 37080

tatggtaaac aaatatgttc tcacagcaag taagatagat gatgtttcat tccttttcca 37140

gaatttagcc cagtgtgaga tttaattgta gggtaccttc ttttctttct tctagatcca 37200

agcataaaag cagacattaa caaatggagg ataaaattgg ataattattc tacccaggaa 37260

ttagggaaaa caattgtcta tctggatggc tcctctcaat catgccgctt tagagaatgc 37320

aacatgggca acctgatttg tgatgcaatg gtaagtcatc agcaggagtg gacatatgct 37380

agggaggaag gaaaggaaga gggaagagga aggaaggatg cgagaaggga tagatcgata 37440

gctacagaat gagggatttc aaaacatttt tagccagggt ggtggcatgt gcctatagtc 37500

ccagctactc gggaggctta ggtgggaaga tggcttgagc ccaggaggct gcagtgagct 37560

ataattgcac cagtgcactc cagcctgggc aaaagagcga gactctgtct ctaaaattaa 37620

ataaatatga aaaataaata aaacattagt gcctaaacaa aagcctgatg tagcttcata 37680

ctcagtacct tcttaaggtg ctggaaggag atcaggtaac caccaacata ctgctttaca 37740

agaactaaaa cctttatttt aaggtgttct tagatgtctg aattttggat gttgctttcc 37800

ttttgtaaga tgatgcaaat gcacaaactc tgctaccctg gagatatcaa cataatagat 37860

aaaagcatat ctgtttggcc agactgtttt tcactcatct gttcacttga ctccagagaa 37920

ccctggcctt gtgtgacagg aacctctttc acaggcacag aaagacaaca cacagtttta 37980

atggtaataa caagtcctat ggctgagcaa gaccaggtat gaagaacaag cccgtctact 38040

tgtcaccagg cggaatggcc atggtagctc tctgggggcc ttactgagcc aaggtctcct 38100

atgtcggact actacttcca tgggctcaca gcctgcaccc agggaagtca tttctgtcag 38160

tgcttggcat gcattgccca gtgcccagga tacaactgca ctcgataaat gtttgccgtg 38220

taaatgacaa caatcatgct catgccaaga ccagactcat gatgtctcct cagcatcatg 38280

cagtcaatgc cagagaagta gttaaggaga agctggtata taaatgttaa gagaagtctg 38340

gagggaaaca acccaggctg ttggcagtgc ttatccactc tgttaaagag gctaggacaa 38400

aggagagctt ttgcttttta tttatttatt tatttaatta attaatttat ttatttattg 38460

agactgagtc ttgctctgtc gcccaggctg gagtgcagtg gcatgatctc agctcactgc 38520

aacctccgtc ccttgggttc aagcgattct cctgcttcag cctcccaagt agctgggatt 38580

agaggtgtcc actaccacac ccagctaatt tttgtatctt tagtagagac ggggtttcac 38640

catgttggct agactggtct cgaactcctg accttaggtg atccacctgc cacagcctcc 38700

caaagtgctg ggatttacag gtgtgagcca ctatgcctgg cctttttttt tttttttttt 38760

cggctttttg tctcctaaat gagaatatat tcattttatt tcttataact aaacatatat 38820

tttgtaaaat aaaagtaaga atggatgtga ttctccccag cttcaaaagg ctgtctctgt 38880

ctttccaaaa ttgccaactt gggaatcagc tgcagacata gacgtggagg ccaggttctc 38940

tttggcatga ggtctggtgt tgagcaggct ctagggccaa gccctcagaa tcaggcagag 39000

ggcatggcct gccctggaga agcctgtgca ggagtgactc tgcaggccac cttgcatggg 39060

aggcccagtc ccgacaccag ggctacaacc gagagaccat tttgctaagg ccacctggcc 39120

tgacagtatg ggagttttag gagggggaaa taagagtttg acatggactg gtaacagtat 39180

atttttactt ctgggagcat aaaattagtt ctaattatag ttgagagatc cctgggaaca 39240

cagcagcagg tgaaaagcag ctttatttcc tcactagagc taaccagatt cctggagctg 39300

atcctaagga agaagagcca gagtaactag tgtaaatctg tgctactgtt gcagattaac 39360

aacaacctga gacacacgga tgaaatgttc tggaaccacg tatccatgtg cattttaaat 39420

ggaggtggta tccggtcgcc cattgatgaa cgcaacaatg gtatgctccc aggcccagct 39480

cctcagtgtg tcatgttctc tctctgatct ccttttctgt tatggttctt gccctgaaca 39540

cacccatgtg cagcctcagt tcatctctgc tgccagccat gtaccagaat gtcagtcctt 39600

taaattaggg ttagccatta ttaaagtcta aaacctagcc tgaaggacac acacaagctt 39660

gtgtgttctt atatgtattc acacacagaa gctgcaggta aggttagtgg aactcaaact 39720

tggtctgaat ttctgactta caaagctata ctagttgttt ctgatttatg caaaataaca 39780

acagtcacca ccttggcatc tgaaaagaga caaaatcacc aatcttctgt cctgaaaaag 39840

acataccact ggattaaaat gttaccttag tatatttaca gatatccaag attatatcat 39900

atggcaaaac aaatcaacta cacaaatact caagacactc ttgtagccct ccctgattta 39960

ctgtgaaaac caatcagtat cagtataagg ccccaaacgc atcccattgt cagacaacag 40020

ggacaggttg caggggcaca ttgtttgtct ggaggaacgg aacaatgcaa aacaaactac 40080

tggagatcca gccccaagct agcagatcta agactgagca tggtgaactt ctgaggcaca 40140

gccatttgtg agctcccgcc atcttgccca gtggtggagt gcagggccag aggcctccta 40200

aactcattcc cttcctcagt gtcaggaacc acttagacat agctaatgat gctctataag 40260

cttcccatgt ccccctcatt tcttccccca gcgctgaggg aaactggtca tggtgtaagc 40320

attttctcaa gctattttcc ttcttgcctc atctgtgact accctcaggc acaattacct 40380

gggagaacct ggctgctgta ttgccctttg gaggcacatt tgacctagtc cagttaaaag 40440

gttccaccct gaagaaggcc tttgagcata gcgtgcaccg ctacggccag tccactggag 40500

agttcctgca ggtgggcggt aagtcaccca tcctgtaggg ctggcccatc caaagtgaca 40560

tggcatttcc tgctggttgg ctcagcttcc ccttcaccaa aattcctgtg gtcaaactgg 40620

tttggatttt tcccgaccca acaccaatac cttacccttt aatggctgtg gactctcaga 40680

gccactaaga ttaggttggg gtcaaagccc caaagcccca aagccctggc aataagcagt 40740

atttgccaag tctgatttgg acatcagatg ggccctctaa gcaggtcacc tgctttgagg 40800

tctagtttct aataaaatct tatttaagca ttcaattatt ttttttttaa atcctctcct 40860

ctgttctact catcgttgcc tggaaccatg ccaagattta aaggaatgct gtccaatctg 40920

tattaaccag gatgattcct ggaaggagca gcctactgca cagacatacc tgaacacctg 40980

gttctagtcc cactgtgacc tggggcaagt tctcaacccc tctgggcaca gggagaaaac 41040

ctttcctgtt tatctcatgg actgcagtga aagttgaatg gaaataaatg cttcctaaat 41100

tgccaagggc ttctgaaaca cttttgttaa taattccaca gtccagtgaa ttaatccttt 41160

aaaagcaatc ttattttacc tggaaatcat aatctggcct cattttttta accttaggtt 41220

gtggtcccgc ttgacagata gaattgcttt cctgagtttt cctctgcaga acacattgct 41280

gttgagactg aagggctaca gaaggcagtg gggccctgtc ctaatgccta tggggcaacc 41340

caaacatcct ccattttccc agaagggagt tgagggaaaa gcagttttct gatgctctag 41400

caaagcactg catagctcta tgccttgtgg aagtcaccaa atcccaaatg attaccattt 41460

aatggtcttc aaaattgtaa aatgctctct gcctctgatc tgtttctcac tttctaaggc 41520

cattgaatca atttagtaga ccaattcagc attttaaggg aaaaacagaa aaagatgtgg 41580

ttttgtccat gtagagtcac cctttgccta ttctggtcca gagctggttt tccttccagt 41640

ggttctcata tgtcctgtaa ggactcaagg ataaagggtg gctgcaatgt cacaattctc 41700

ttgacaaaat ttcactcagg tttaaaccaa aggaaaaacc acagccccat gccccaattg 41760

tagagtttgg ccaaaattcc ttaggaaaca gaaatctccc tttggatctg gtgaaaacag 41820

attcatttct tttctaggaa tccatgtggt gtatgatctt tcccgaaaac ctggagacag 41880

agtagtcaaa ttagatgttc tttgcaccaa gtgtcgagtg cccagttatg accctctcaa 41940

aatggacgag gtatataagg tgatcctccc aaacttcctg gccaatggtg gagatgggtt 42000

ccagatgata aaagatgaat tattaagaca tgactctggt aagcatgact gtctcttcct 42060

ttctctaaag aacaacaaaa ttgggccaag aaggggagct actagtggtg ccaggagggc 42120

aaagtgactc cctgtattgc tgactccctc ctgctttgct tgttggaaag cagcagcaca 42180

gcacagcaga gaggaatatg taccctgtca atccccgagc acacacatgc cttcatccaa 42240

ccccagagac aaaacatttg taaatcaaaa atgttttaaa attcttttaa ctacatctaa 42300

gcagctattt aaattgtgaa atataagtga gttttgcatt aacagaaata tcacacctct 42360

ggaattttag gatatggaaa accatcacat taaatcattt aaattttctt tcttgaagaa 42420

gtttctgtgt gaatcagtca ggtagaagtg gatggggtga gcgccgccct ttcagcacag 42480

tgggttggtg gtacaaatca taggaataac catacttgat gtcagttggt acttctggca 42540

gcattgaggg acattatatg ccagcctcac ttttagaaat tctgttgaga gtcacattat 42600

gtgcatacag accactgaca taggacacac ttccccttcc tcttcctagg gcagagacca 42660

gcatgcttac aagacacaca agcagcttct tggcatctct gggtagttcc accaggatct 42720

tcctcagtct tatcatgctg gacctgtcat tttagctact tattccaagg agcgtggagg 42780

gcaggaccag accaaaggag gcagcccttc ctgggaggaa aggtgctggc ttcttctgtc 42840

ccttccgtga cagaagatgg aagggcaggc gtaaaagccc catgagacaa agaggctgat 42900

agccatcagc tgtccaccac tcaagcctag actgaaggat gttagagaga ggagctcatg 42960

cagaaggcta aggaagccct ggacctaagc gttctcaaca ctggttatac attagaatca 43020

cttcaggagc attttaaatt aaataccaat ggctcggccg cagctcttaa gagattctaa 43080

gttaactgga ttggggctaa actctttttc agtgattcta gtgtacaccc agaggtgaaa 43140

accactgaag agagaagggt tcttaagtac tggagtgata tttttaaaat aaaattctgt 43200

tgggttccca acagaacatg ggttcccaac aaacatgggt ttgttgtagc tataggcaat 43260

tagcatttat cctctcctct ctttggtaaa ggaaatcccc caagtcccat catcaattac 43320

atgaaaataa ttattgggca ggcaaacaaa aggaagccag ttttatctct caactaattg 43380

ctcttgaccc agttccctaa tgcccttgtt ctggagtggt tcaaaggcta tgatgtcaat 43440

gacctctttc tagagtcatc acagagctga ggcaactcta cttcccccaa ttacccaata 43500

ttgaacaaca gacacatttt acatctggag catgaaaaag caatcaaatt ctttgtactt 43560

atgatcacta gcgttcttgt cttctgtgac actttacccc tgcttatgaa attgcttccc 43620

ttttataatt acaaaggact accttactgt tgattgataa ttaccttttc tgtagttaaa 43680

ataatgtata tgtttttcta ggtgaccaag atatcaacgt ggtttctaca tatatctcca 43740

aaatgaaagt aatttatcca gcagttgaag gtcggatcaa gttttccaca ggaagtcact 43800

gccatggaag cttttcttta atatttcttt cactttgggc agtgatcttt gttttatacc 43860

aatagccaaa aattctcctt gcctttaatg tgtgaaactg cattttttca agtgagattc 43920

aaatctgcct tttaggacct ggctttgtga cagcaaaaac catctttaca ggctcctaga 43980

agctgaaggt tagagcatta taaaatgaag agacagacat gattactcag ggtcagcaac 44040

ctagtgagtt agaaaaaaaa ttaacatagg gccctataag gagaaagcca actatgttaa 44100

gtttacgtgt ccaaatttta atgaaatttt actaacaatt ttaaaccata tttttcttct 44160

tcatatccat ttctaatcca tcaaacagct tatgtttaca taaaatttta tcattcacaa 44220

ggaagtttta agcacactgt ctcatttgat atccacaact tatttttggt aggaaagaga 44280

gatgtttttc ccacctgtca gatgaaaaaa ctgaagctca aaaagggttg acttgaccat 44340

acagctaatg ctgacagatc caagacctag acctaggtct tttgaactca agtccagcat 44400

tctcaactat atcaagttac tgttcagaat acttaatatc tcctctcttc ataattatca 44460

atagccccaa gctcatggat gacaaatctc tgctttattt cttgtctcta ttttttcact 44520

ttatagctcc tgttataata gcaagtttaa tggtataaac acaggatacc atcctctctt 44580

gcaacaccca tgtgcctttg atgagtcagg tagcaagctg tagtagataa tgagaaaggc 44640

cagaggctgc aaaagacagt caaaggacac gagagaaagg aaggggaaga acaggactcc 44700

aggactgttt tatattatag aaaagcaaga gctaaagagc atttacacat gttaaacaga 44760

tacttgttaa gcatagtgcc tgacacacgg cattagctgt tattttatga gattccatca 44820

gctctgcctc tgtcctcttt cttctaacat gaaggtatca tgagaagaga accttctaac 44880

ataagctgta attctaaacc tgcacttgtc cctctccagc aagaggctag cactgaattc 44940

attctactca tactacacac ccagttatgg aatgtccaga gttctcgaag aaaataaatg 45000

actttaggaa gaggtataca ttttttaagt cgctctgcct ccaaatctga acagtcactg 45060

taaatcattc ttaagcccag atatgagaac ttctgctgga aagtgggacc ctctgagtgg 45120

gtggtcagaa aatacccatg ctgatgaaat gacctatgcc caaagaacaa atacttaacg 45180

tgggagtgga accacatgag cctgctcagc tctgcataag taattcaaga aatgggaggc 45240

ttcaccttaa aaacagtgtg caaatggcag ctagaggttt tgataggaag tatgtttgtt 45300

tcttagtgtt tacaaatatt aagtactctt gatacaaaat atacttttaa acttcataac 45360

ctttttataa aagttgttgc agcaaaataa tagcctcggt tctatgcata tatggattag 45420

ctataaaaaa tgtcaataag attgtacaag gaaaattaga gaaagtcaca tttagggttt 45480

attttttaca cttggccagt aaaatagggt aaatcctatt agaatttttt aaagaacttt 45540

ttttaagttt cctaaatctg tgtgtgtatt gtgaagtggt ataagaaatg actttgaacc 45600

actttgcaat tgtagattcc caacaataaa attgaagata agctctttgg tctttgatag 45660

ttatttcttc tctcattcaa acacctgcca actaaagaca atatgactga aggaaaactc 45720

tgacagattt tcactggtat catttctctt gatcataagc cacttaccat ctgataaccc 45780

tccatagccc ctcacacccc taaaatgaga agaaaaatgc atccaatttt taagtaggag 45840

agaaaagtga gtggtatttt aaggaagact actaaatttg gtggctttga tatacagaat 45900

ctgatataaa aatggttttg aggccagtat aacttcgtat aatgtatgct atacgaagtt 45960

ataggtctga agaggagttt acgtccagcc aagctagcgg ctccggtgcc cgtcagtggg 46020

cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaaccg 46080

gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc 46140

tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt 46200

ttcgcaacgg gtttgccgcc agaacacagg taagtgccgt gtgtggttcc cgcgggcctg 46260

gcctctttac gggttatggc ccttgcgtgc cttgaattac ttccacctgg ctccagtacg 46320

tgattcttga tcccgagctg gagccagggg cgggccttgc gctttaggag ccccttcgcc 46380

tcgtgcttga gttgaggcct ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca 46440

ccttcgcgcc tgtctcgctg ctttcgataa gtctctagcc atttaaaatt tttgatgacc 46500

tgctgcgacg ctttttttct ggcaagatag tcttgtaaat gcgggccagg atctgcacac 46560

tggtatttcg gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg 46620

ttcggcgagg cggggcctgc gagcgcggcc accgagaatc ggacgggggt agtctcaagc 46680

tggccggcct gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc 46740

aaggctggcc cggtcggcac cagttgcgtg agcggaaaga tggccgcttc ccggccctgc 46800

tccagggggc tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg agtcacccac 46860

acaaaggaaa ggggcctttc cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg 46920

ggcgccgtcc aggcacctcg attagttctg gagcttttgg agtacgtcgt ctttaggttg 46980

gggggagggg ttttatgcga tggagtttcc ccacactgag tgggtggaga ctgaagttag 47040

gccagcttgg cacttgatgt aattctcctt ggaatttgcc ctttttgagt ttggatcttg 47100

gttcattctc aagcctcaga cagtggttca aagttttttt cttccatttc aggtgtcgtg 47160

agcggccgga agcttcacga ggccgccacc cccggggttg acaattaatc atcggcatag 47220

tatatcggca tagtataata cgacaaggtg aggaactaaa ccatgggatc ggccattgaa 47280

caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac 47340

tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg 47400

cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag 47460

gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt 47520

gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg 47580

tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg 47640

catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga 47700

gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag 47760

gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgatgat 47820

ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt 47880

tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg 47940

gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt 48000

tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc 48060

ttctgagggg atcaattctc tagagctcgc tgatcagcct cgactgtgcc ttctagttgc 48120

cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc 48180

actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct 48240

attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg 48300

catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctcg 48360

actagagctt gcggaaccct tc 48382

<210> 19

<211> 3582

<212> DNA/RNA

<213> Artificial Sequence (Artificial Sequence)

<400> 19

agtttagtag aggccccggt ggccggtgct cactgggctc agggccagtc cacccgctca 60

acgcgctcta gccagccatg tgtccccgag ccgcgcgggc gcccgcgacg ctactcctcg 120

ccctgggcgc ggtgctgtgg cctgcggctg gcgcctggga gcttacgatt ttgcacacca 180

acgacgtgca cagccggctg gagcagacca gcgaggactc cagcaagtgc gtcaacgcca 240

gccgctgcat gggtggcgtg gctcggctct tcaccaaggt tcagcagatc cgccgcgccg 300

aacccaacgt gctgctgctg gacgccggcg accagtacca gggcactatc tggttcaccg 360

tgtacaaggg cgccgaggtg gcgcacttca tgaacgccct gcgctacgat gccatggcac 420

tgggaaatca tgaatttgat aatggtgtgg aaggactgat cgagccactc ctcaaagagg 480

ccaaatttcc aattctgagt gcaaacatta aagcaaaggg gccactagca tctcaaatat 540

caggacttta tttgccatat aaagttcttc ctgttggtga tgaagttgtg ggaatcgttg 600

gatacacttc caaagaaacc ccttttctct caaatccagg gacaaattta gtgtttgaag 660

atgaaatcac tgcattacaa cctgaagtag ataagttaaa aactctaaat gtgaacaaaa 720

ttattgcact gggacattcg ggttttgaaa tggataaact catcgctcag aaagtgaggg 780

gtgtggacgt cgtggtggga ggacactcca acacatttct ttacacaggc aatccacctt 840

ccaaagaggt gcctgctggg aagtacccat tcatagtcac ttctgatgat gggcggaagg 900

ttcctgtagt ccaggcctat gcttttggca aatacctagg ctatctgaag atcgagtttg 960

atgaaagagg aaacgtcatc tcttcccatg gaaatcccat tcttctaaac agcagcattc 1020

ctgaagatcc aagcataaaa gcagacatta acaaatggag gataaaattg gataattatt 1080

ctacccagga attagggaaa acaattgtct atctggatgg ctcctctcaa tcatgccgct 1140

ttagagaatg caacatgggc aacctgattt gtgatgcaat gattaacaac aacctgagac 1200

acacggatga aatgttctgg aaccacgtat ccatgtgcat tttaaatgga ggtggtatcc 1260

ggtcgcccat tgatgaacgc aacaatggca caattacctg ggagaacctg gctgctgtat 1320

tgccctttgg aggcacattt gacctagtcc agttaaaagg ttccaccctg aagaaggcct 1380

ttgagcatag cgtgcaccgc tacggccagt ccactggaga gttcctgcag gtgggcggaa 1440

tccatgtggt gtatgatctt tcccgaaaac ctggagacag agtagtcaaa ttagatgttc 1500

tttgcaccaa gtgtcgagtg cccagttatg accctctcaa aatggacgag gtatataagg 1560

tgatcctccc aaacttcctg gccaatggtg gagatgggtt ccagatgata aaagatgaat 1620

tattaagaca tgactctggt gaccaagata tcaacgtggt ttctacatat atctccaaaa 1680

tgaaagtaat ttatccagca gttgaaggtc ggatcaagtt ttccacagga agtcactgcc 1740

atggaagctt ttctttaata tttctttcac tttgggcagt gatctttgtt ttataccaat 1800

agccaaaaat tctccttgcc tttaatgtgt gaaactgcat tttttcaagt gagattcaaa 1860

tctgcctttt aggacctggc tttgtgacag caaaaaccat ctttacaggc tcctagaagc 1920

tgaaggttag agcattataa aatgaagaga cagacatgat tactcagggt cagcaaccta 1980

gtgagttaga aaaaaaatta acatagggcc ctataaggag aaagccaact atgttaagtt 2040

tacgtgtcca aattttaatg aaattttact aacaatttta aaccatattt ttcttcttca 2100

tatccatttc taatccatca aacagcttat gtttacataa aattttatca ttcacaagga 2160

agttttaagc acactgtctc atttgatatc cacaacttat ttttggtagg aaagagagat 2220

gtttttccca cctgtcagat gaaaaaactg aagctcaaaa agggttgact tgaccataca 2280

gctaatgctg acagatccaa gacctagacc taggtctttt gaactcaagt ccagcattct 2340

caactatatc aagttactgt tcagaatact taatatctcc tctcttcata attatcaata 2400

gccccaagct catggatgac aaatctctgc tttatttctt gtctctattt tttcacttta 2460

tagctcctgt tataatagca agtttaatgg tataaacaca ggataccatc ctctcttgca 2520

acacccatgt gcctttgatg agtcaggtag caagctgtag tagataatga gaaaggccag 2580

aggctgcaaa agacagtcaa aggacacgag agaaaggaag gggaagaaca ggactccagg 2640

actgttttat attatagaaa agcaagagct aaagagcatt tacacatgtt aaacagatac 2700

ttgttaagca tagtgcctga cacacggcat tagctgttat tttatgagat tccatcagct 2760

ctgcctctgt cctctttctt ctaacatgaa ggtatcatga gaagagaacc ttctaacata 2820

agctgtaatt ctaaacctgc acttgtccct ctccagcaag aggctagcac tgaattcatt 2880

ctactcatac tacacaccca gttatggaat gtccagagtt ctcgaagaaa ataaatgact 2940

ttaggaagag gtatacattt tttaagtcgc tctgcctcca aatctgaaca gtcactgtaa 3000

atcattctta agcccagata tgagaacttc tgctggaaag tgggaccctc tgagtgggtg 3060

gtcagaaaat acccatgctg atgaaatgac ctatgcccaa agaacaaata cttaacgtgg 3120

gagtggaacc acatgagcct gctcagctct gcataagtaa ttcaagaaat gggaggcttc 3180

accttaaaaa cagtgtgcaa atggcagcta gaggttttga taggaagtat gtttgtttct 3240

tagtgtttac aaatattaag tactcttgat acaaaatata cttttaaact tcataacctt 3300

tttataaaag ttgttgcagc aaaataatag cctcggttct atgcatatat ggattagcta 3360

taaaaaatgt caataagatt gtacaaggaa aattagagaa agtcacattt agggtttatt 3420

ttttacactt ggccagtaaa atagggtaaa tcctattaga attttttaaa gaactttttt 3480

taagtttcct aaatctgtgt gtgtattgtg aagtggtata agaaatgact ttgaaccact 3540

ttgcaattgt agattcccaa caataaaatt gaagataagc tc 3582

<210> 20

<211> 1349

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 20

attaagggtt ccggatcctc ggggacacca aatatggcga tctcggcctt ttcgtttctt 60

ggagctggga catgtttgcc atcgatccat ctaccaccag aacggccgtt agatctgctg 120

ccaccgttgt ttccaccgaa gaaaccaccg ttgccgtaac caccacgacg gttgttgcta 180

aagaagctgc caccgccacg gccaccgttg tagccgccgt tgttgttatt gtagttgctc 240

atgttatttc tggcacttct tggttttcct cttaagtgag gaggaacata accattctcg 300

ttgttgtcgt tgatgcttaa attttgcact tgttcgctca gttcagccat aatatgaaat 360

gcttttcttg ttgttcttac ggaataccac ttgccaccta tcaccacaac taactttttc 420

ccgttcctcc atctctttta tatttttttt ctcgagggat ctttgtgaag gaaccttact 480

tctgtggtgt gacataattg gacaaactac ctacagagat ttaaagctct aaggtaaata 540

taaaattttt aagtgtataa tgtgttaaac tactgattct aattgtttgt gtattttaga 600

ttccaaccta tggaactgat gaatgggagc agtggtggaa tgcctttaat gaggaaaacc 660

tgttttgctc agaagaaatg ccatctagtg atgatgaggc tactgctgac tctcaacatt 720

ctactcctcc aaaaaagaag agaaaggtag aagaccccaa ggactttcct tcagaattgc 780

taagtttttt gagtcatgct gtgtttagta atagaactct tgcttgcttt gctatttaca 840

ccacaaagga aaaagctgca ctgctataca agaaaattat ggaaaaatat tctgtaacct 900

ttataagtag gcataacagt tataatcata acatactgtt ttttcttact ccacacaggc 960

atagagtgtc tgctattaat aactatgctc aaaaattgtg tacctttagc tttttaattt 1020

gtaaaggggt taataaggaa tatttgatgt atagtgcctt gactagagat cataatcagc 1080

cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac 1140

ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt 1200

tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct 1260

agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatctg acatggtaag 1320

taagcttggg ctgcaggtcg agggaccta 1349

<210> 21

<211> 132

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 21

gaattctaat acgactcact atagggggtc ttcgagaaga cctgttttag agctagaaat 60

agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct 120

tttaaaggat cc 132

<210> 22

<211> 1865

<212> DNA/RNA

<213> Artificial Sequence (Artificial Sequence)

<400> 22

cagggagtct ccttgtcctt gatgtcaaac tacatttttc ttccagtgat attcatatct 60

gcctctggag acctggcttt gtaacagcac tcatcatcct caaggttcct agcagatgtt 120

cttcacaagg aagagactgt aacatcattt gttggggcca gcaactcagt gagcagatag 180

aaagtcacag tgaaccaaca gggtccttct ggcagggagt gggtaagggg aaacaactag 240

atgtagcttg catagccaca taacacatct ggttaccatt tccctttcta ttcatttcta 300

atccatcaaa caattgatgt ttacatacaa cttcatcatc gccagctctg gtggcacatg 360

cctgtggtca cggcacttgg cagggaggag aggatggctg caagttctag gccagcctga 420

catatgtaga gtttcaggcc agtcagctag atatcaagac tcacacacac aaacaaacat 480

tataatttac aagtagattt ttgtagacaa gtcttatgat aagctaagca gaaagggtcg 540

acttgttcaa ggtcagacat ctccaaataa tacatctaag atctgtacct gagtcttttt 600

acctcaagtc caacctctcg cctactgtca agtctcctct cttcctgcgg ttggtctcag 660

acaacaaatc tgtttctgcc ttccatagta ccctttcttt tgggctcttg ttgtctctca 720

ggtttgagag agtagctact ggacaggatt ctttcctaca acaccatgtg cctttggtga 780

gtcatagaca aactgtatac agcagataat agattagtcc agggactgca aaaggcagtc 840

agggacaggc aggaaaggga aaggagaata tgaccaggac ttacagaaga gagtagatgg 900

gttccataaa cactagttaa ggagcagaac atggctccaa agcacaacac tggttcttat 960

taactcctgt gcgactccaa aagccctgcc tctctccttc cttcttcctc acatggaggt 1020

gccatgagag aacttcctaa aataaactgt aattctcaac ctgcacctgt cctgtccaag 1080

atgccaacac ggctctcatt caagtcacac atgtacaatc caatattctg gaaggcaaga 1140

gtgaatttaa gaagtggtat acataaatgt ttgaagtcac tgagactcaa atcccacaca 1200

accactgtaa ggcatactca ggtcaagaca tgagaagacc agcaggacag tgagccctat 1260

gggaggccca agagataccc agactgatga aatgatctgt gcccacagaa caccagacca 1320

agacagtcct gtgaccaagt gagcatagtc agcactgtac agtagttaag aatccttaac 1380

tataaaccag cagctagatg tctggacagg gaacatgttg gtttcctagt gtttacgaat 1440

attaaggact cttgacccaa accgtttaaa attccaatcc ttttatgaag tttgttgcag 1500

tgaaacaatg ccttcagttc tttttgaatg tgtagattag ttataaacag aaatgtcagt 1560

aagactgtaa aaaaaaatga agaaaggtgc atatgaggta tatttttatg cttggcgagt 1620

aaataaggag aaatcttata gcataatttt taaagaacat ttgtataatt tttctaaata 1680

tgtgtatatg tatattttat gcagcagtat tgagaaatga ccttggacta tcttgtaatt 1740

gtaaattcca aaaataaagc tgaagatata gtcctttgtc tttggtctta cttcttctct 1800

taacacttgt accctcaggg atgcagcaaa gacaatgttc acttcaaaac tgagacgaac 1860

aatgc 1865

<210> 23

<211> 90

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 23

gcggaaccct tcgaagttcc tattctctag aaagtatagg aacttcccag agcccaaggc 60

actcattatt agcactgggt gagctccatg 90

<210> 24

<211> 25

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 24

gagtggtatg tgcagctcag ggaag 25

<210> 25

<211> 25

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 25

gctccaagaa acgaaaaggc cgaga 25

<210> 26

<211> 23

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 26

agcgcaaaca ttaaggcacg ggg 23

<210> 27

<211> 23

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 27

ttaatgtttg cgctcagaat tgg 23

<210> 28

<211> 23

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 28

gagcgcaaac attaaggcac ggg 23

<210> 29

<211> 23

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 29

aattctgagc gcaaacatta agg 23

<210> 30

<211> 23

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 30

cctctagcac atcagatatc tgg 23

<210> 31

<211> 23

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 31

tgagcgcaaa cattaaggca cgg 23

<210> 32

<211> 23

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 32

gattgttgga tatacttcaa agg 23

<210> 33

<211> 23

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 33

ttggaaattt aacgtttctg agg 23

<210> 34

<211> 23

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 34

catcacttgc ggggaagtcc tgg 23

<210> 35

<211> 23

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 35

tgctgagtcc atcacttgcg ggg 23

<210> 36

<211> 23

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 36

caggacttcc ccgcaagtga tgg 23

<210> 37

<211> 23

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 37

gtctgtaacc ccgataagct tgg 23

<210> 38

<211> 23

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 38

ttgcatctgc ggagttagca tgg 23

<210> 39

<211> 23

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 39

gccatcgacc attcgtgcac agg 23

<210> 40

<211> 23

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 40

ccctgtgcac gaatggtcga tgg 23

<210> 41

<211> 23

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 41

agtggagccc tgtgcacgaa tgg 23

<210> 42

<211> 23

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 42

cctttgaagt atatccaaca atc 23

<210> 43

<211> 27

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 43

tagggattgt tggatatact tcaaagg 27

<210> 44

<211> 27

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 44

aaaccctttg aagtatatcc aacaatc 27

<210> 45

<211> 23

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 45

cctgtgcacg aatggtcgat ggc 23

<210> 46

<211> 27

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 46

tagggccatc gaccattcgt gcacagg 27

<210> 47

<211> 27

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 47

aaaccctgtg cacgaatggt cgatggc 27

<210> 48

<211> 26

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 48

tttgataacg gtgtggaagg actgat 26

<210> 49

<211> 25

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 49

agacagcgat gcccttcaga ggata 25

<210> 50

<211> 24

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 50

tacacggtga accagatagt gccc 24

<210> 51

<211> 25

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 51

gtgggcggaa tccatgtggt gtatg 25

<210> 52

<211> 25

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 52

gccccagggc gatgatctta ttcac 25

<210> 53

<211> 25

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 53

agagaccctt tagacctgga tcagc 25

<210> 54

<211> 26

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 54

acctagaaaa ctttggtggg acatga 26

<210> 55

<211> 25

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 55

aactgatgaa tgggagcagt ggtgg 25

<210> 56

<211> 25

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 56

gcagacactc tatgcctgtg tggag 25

<210> 57

<211> 25

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 57

aagatgtttc ttgtcccggg tttcg 25

<210> 58

<211> 25

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 58

ggactgattg atcccctcct cagaa 25

<210> 59

<211> 22

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 59

gcacctgccg tccatcatct gc 22

<210> 60

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 60

acgtgctgtt tttggatgcc 20

<210> 61

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 61

tggtgtgcaa aatcgtaagc 20

<210> 62

<211> 22

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 62

tcaccatctt ccaggagcga ga 22

<210> 63

<211> 21

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 63

gaaggccatg ccagtgagct t 21

Claims

1. A construction method of a non-human animal humanized with a CD73 gene is characterized in that the genome of the non-human animal comprises a human or humanized CD73 gene, and the non-human animal expresses a human or humanized CD73 protein.

2. The method of claim 1, wherein the amino acid sequence of the humanized CD73 protein comprises one of the following groups:

3. Construction method according to claim 1 or 2, wherein the humanized CD73 gene comprises all or part of exons 1 to 9 of human CD73 gene, preferably comprises all or part of exons 1,2 to 8 and 9, wherein part of exons 1 comprises at least 100bp of nucleotide sequence and part of exons 9 comprises at least 50bp of nucleotide sequence, and more preferably the humanized CD73 gene comprises all or part of CDs sequence of human CD73 gene or all or part of full-length nucleotide sequence of human CD 73.

4. The construct of any one of claims 1-3, wherein the humanized CD73 gene comprises one of the following groups:

5. The construct of any one of claims 1-4, wherein the humanized CD73 gene comprises one of the following groups:

(iii) comprises a nucleotide sequence substantially identical to SEQ ID NO: 12. 13 or 18 differ by no more than 10, 9, 8, 7, 6, 5, 4, 3, 2, or no more than 1 nucleotide;

(iv) comprises a nucleotide sequence substantially identical to SEQ ID NO: 12. 13 or 18, including nucleotide sequences with one or more nucleotides substituted, deleted and/or inserted;

(v) the transcribed mRNA comprises SEQ ID NO:19, or a portion or all of the nucleotide sequence set forth in seq id no;

(vi) the transcribed mRNA comprises a sequence identical to SEQ ID NO:19 is at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or at least 99% identical;

(vii) the transcribed mRNA comprises a sequence identical to SEQ ID NO:19 by no more than 10, 9, 8, 7, 6, 5, 4, 3, 2, or by no more than 1 nucleotide; or the like, or, alternatively,

(viii) the transcribed mRNA comprises a sequence identical to SEQ ID NO:19, including nucleotide sequences with one or more nucleotides substituted, deleted and/or inserted.

6. The method of construction according to any one of claims 1 to 5, comprising introducing a nucleotide sequence comprising a human or humanized CD73 gene into the non-human animal CD73 locus, preferably into the non-human animal CD73 locus, preferably comprising all or part of exons 1 to 9 of the human CD73 gene, further preferably comprising all or part of exon 1, all of exons 2 to 8, and all or part of exons 9, into the non-human animal CD73 locus, wherein part of exon 1 comprises at least a nucleotide sequence of 100bp, part of exon 9 comprises at least a nucleotide sequence of 50bp, further preferably comprises all or part of a CDS sequence of the human CD73 gene or all or part of a full-length nucleotide sequence of human CD73 into the non-human animal CD73 locus, further preferably, the construction method comprises the following steps of using a polypeptide containing a nucleotide sequence encoding SEQ ID NO:4, and yet further preferably comprises a nucleotide sequence comprising SEQ ID NO: 9. 12, 13, 17 or 18 into the CD73 locus of a non-human animal.

7. The method of constructing according to any one of claims 1 to 6, wherein the genome of the non-human animal further comprises other genetic modifications, and more preferably, the other genes comprise at least one of PD-1, PD-L1, CTLA-4, LAG-3, BTLA, CD27, CD28, CD47, CD137, CD154, OX40, SIRPa, TIGIT, TIM-3, CD40, and GITR genes.

8. A targeting vector comprising a portion of the human CD73 gene, preferably the targeting vector comprises all or part of exons 1 to 9, preferably all or part of exons 1,2 to 8 and 9 of the human CD73 gene, wherein the portion of exons 1 comprises at least 100bp of nucleotide sequence, the portion of exons 9 comprises at least 50bp of nucleotide sequence, more preferably all or part of the CDs sequence of the human CD73 gene or all or part of the full-length nucleotide sequence of human CD73, even more preferably all or part of the CDs sequence of the human CD73 gene or all or part of the full-length nucleotide sequence of human CD73, yet more preferably the targeting vector comprises a polynucleotide encoding a polypeptide of SEQ ID NO:4, further preferably comprises the nucleotide sequence of SEQ ID NO: 9. 12, 13, 17 or 18.

9. The targeting vector according to claim 8, wherein said targeting vector further comprises a 5' arm and/or a3 ' arm, said 5' arm is selected from the group consisting of nucleotides of 100-10000 in length of the genomic DNA of the non-human animal CD73 gene; preferably, said 5' arm has at least 90% homology to NCBI accession No. NC _ 000075.6; further preferably, the 5' arm sequence is identical to SEQ ID NO: 5. 7 or 15 have at least 90% homology; the 3' arm is selected from 100-10000 nucleotides in length of the non-human animal CD73 gene genome DNA; preferably, said 3' arm has at least 90% homology to NCBI accession No. NC _ 000075.6; further preferably, the 3' arm sequence is identical to SEQ ID NO: 6. 8 or 16 have at least 90% homology.

10. A humanized CD73 gene, wherein the humanized CD73 gene comprises a portion of the human CD73 gene.

11. The humanized CD73 gene according to claim 10, wherein the humanized CD73 gene comprises all or part of exons 1 to 9 of human CD73 gene, preferably comprises all or part of exons 1,2 to 8 and 9, wherein part of exons 1 comprises at least 100bp of nucleotide sequence and part of exons 9 comprises at least 50bp of nucleotide sequence, more preferably comprises all or part of CDs sequence of human CD73 gene or all or part of full-length nucleotide sequence of human CD 73.

12. The humanized CD73 gene of claim 10 or 11, wherein the humanized CD73 gene comprises one of the following group:

13. A cell, tissue or organ derived from a non-human animal obtained by the method of construction of any one of claims 1 to 7, wherein the cell, tissue or organ comprises the humanized CD73 gene of any one of claims 10 to 12, and wherein the cell, tissue or organ expresses a human or humanized CD73 protein.

14. Use of a cell, tissue or organ derived from a non-human animal obtained by the method of construction according to any one of claims 1 to 7, the humanized CD73 gene according to any one of claims 10 to 12, or the cell, tissue or organ according to claim 13 for product development of an immune process involving human cells, for production of antibodies, or as a model system for pharmacological, immunological, microbiological or medical research; or in the production and use of animal experimental disease models for the development of new diagnostic and/or therapeutic strategies; or screening, verifying, evaluating or researching CD73 channel function, human CD73 channel signal mechanism, human-targeting antibody, human-targeting medicine, medicine effect, immunity-related disease medicine and anti-tumor medicine, screening and evaluating human medicine and medicine effect research.