CN108504685A - A method of utilizing CRISPR/Cas9 system homologous recombination repair IL-2RG dcc genes - Google Patents

A method of utilizing CRISPR/Cas9 system homologous recombination repair IL-2RG dcc genes Download PDF

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CN108504685A
CN108504685A CN201810260425.4A CN201810260425A CN108504685A CN 108504685 A CN108504685 A CN 108504685A CN 201810260425 A CN201810260425 A CN 201810260425A CN 108504685 A CN108504685 A CN 108504685A
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paav
carriers
aavs1
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sacas9
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孙秀莲
吴向萍
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Immune Cell Biotechnology Co Ltd
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Immune Cell Biotechnology Co Ltd
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    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • C12N15/8509—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells for producing genetically modified animals, e.g. transgenic
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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    • A01K67/00—Rearing or breeding animals, not otherwise provided for; New breeds of animals
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    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/715—Receptors; Cell surface antigens; Cell surface determinants for cytokines; for lymphokines; for interferons
    • C07K14/7155—Receptors; Cell surface antigens; Cell surface determinants for cytokines; for lymphokines; for interferons for interleukins [IL]
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    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
    • C12N15/90—Stable introduction of foreign DNA into chromosome
    • C12N15/902—Stable introduction of foreign DNA into chromosome using homologous recombination
    • C12N15/907—Stable introduction of foreign DNA into chromosome using homologous recombination in mammalian cells
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    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
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    • C12N9/22—Ribonucleases RNAses, DNAses
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2217/00—Genetically modified animals
    • A01K2217/07—Animals genetically altered by homologous recombination
    • A01K2217/072—Animals genetically altered by homologous recombination maintaining or altering function, i.e. knock in
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2227/00—Animals characterised by species
    • A01K2227/10—Mammal
    • A01K2227/105—Murine
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2267/00—Animals characterised by purpose
    • A01K2267/03—Animal model, e.g. for test or diseases
    • A01K2267/035—Animal model for multifactorial diseases
    • A01K2267/0387—Animal model for diseases of the immune system

Abstract

The invention discloses a kind of methods using CRISPR/Cas9 system homologous recombination repair IL 2RG dcc genes,By obtaining pAAV AAVS1Donor carriers and pAAV SaCas9 gRNAAAVS1 carriers for repairing IL 2RG dcc genes,By will be after its co-implanted X SCID host cell,CRISPR/Cas9 systems in pAAV SaCas9 gRNA AAVS1 carriers cut defect IL 2RG genes in host cell,And correct IL 2RG genes in pAAV AAVS1Donor carriers are properly inserted in genome using homologous recombination repair mechanism,The method that the present invention utilizes CRISPRCas9 system homologous recombination repair IL 2RG dcc genes,Realize the breakthrough that X SCID are precisely treated.

Description

It is a kind of using CRISPR/Cas9 system homologous recombination repair IL-2RG dcc genes Method
Technical field
The invention belongs to genetic engineering fields, and in particular to a kind of to utilize CRISPR/Cas9 system homologous recombination repairs IL- The method of 2RG dcc genes.
Background technology
Primary immunodeficiency disease (Primary immunodeficiency diseases, PID) is because immune system is lost Defect or congenital development is passed not cause to be susceptible to suffer from infection, malignant tumour and autoimmune disease caused by immune dysfunction entirely One group of disease has had now been found that the mutation of 200 several genes leads to PID, and has been sent out per having more than ten to plant new PID diseases every year Existing, although single PID disease incidence is low, the state of an illness is critical, poor prognosis, and whole incidence is not low, and China is newly sent out every year PID infants are up to thousands of examples.
Severe combined immunodeficiency (severe combined immunodeficiency, SCID) is the one of most serious Kind PID types, are one group of diseases for seriously threatening children's health, and developed country has been included in the range of neonatal screening, SCID is caused with T alymphocytosises or dysfunction, companion or without bone-marrow-derived lymphocyte and NK cells by several genes defect One group of disease that quantity is reduced or functional defect is characterized, wherein about 50% is X linked severe combined immunodeficiency diseases (X- SCID), it is the hereditary immunodeficiency disease that life is seriously threatened caused by common γ chains (IL-2RG) gene mutation of IL-2 receptors, Most infants are dead within 1 years old, and hematopoietic stem cell transplantation is the current main radical treatment hands of X-SCID in the world Section, but severe infection before being sought and transplanted due to the donor difficulty of complete distribution type include the BCG epidemic diseases of the legal neonatal period inoculation in China Seedling infects and the reasons such as graft versus host disease(GVH disease) (graft-versus-host disease, GVHD) cause China's Hematopoietic Stem thin The success rate of born of the same parents' transplanting is extremely low, and the data that the whole nation has not been reported shows that the left and right only about 5%-10% (1-2/13) X-SCID infants are logical It crosses hematopoietic stem cell transplantation to survive, gene therapy solves distribution type problem, the incidence of graft versus host disease(GVH disease) after treatment Greatly reduce.
Chinese patent CN201610819040.8 discloses a kind of congenital immune deficiency gene therapy vector construction method And application thereof, the SIN-LV-IL2RG carriers with high promoter activity are constructed, are carried using the higher SIN-LV of safety Body, the SINLV viral vectors that structure IL-2RG is overexpressed, and SIN-LV is overexpressed IL-2RG infected rats models (IL-2RG Knockout rat model), changed by the symptom of analyzing rat model, the validity of gene therapy X-SCID can be made Accurately assessment, but the immunogenicity of viral vectors is strong, internal expression time is short and the tumorigenic risk meeting of radom insertion Hinder the extensive use of clinical gene therapy.
Gene editing technology refers to allow the mankind to carry out " editor " to target gene, realization to the knockouts of specific DNA fragments, Be added etc., and CRISPR/Cas9 technology since the advent of the world, just there are other unrivaled advantages of gene editing technology, technology After continuously improving, be more believed in living cells it is most effective, most easily " edit " any gene, CRISPRCas9 technologies To combining target DNA sequence dna and fixed point cutting can be carried out by guide RNA targets, pass through homologous recombination repair (homology- Directed repair, HDR) or non-homologous end joining (NHEJ) repaired at gene break, therefore improve base Because of the specificity and efficiency of editor, hope is brought for the accurate treatment of genetic disease, especially X-SCID.
Currently, CRISPR/Cas9 systematic differences mainly have following example：
For example, Chinese patent CN201510198437.5 utilizes CRISPR/CAS9 genome editing techniques, the structure mankind to lose The disease cardiac muscle cell of transmissibility cardiomyopathy multipotential stem cell, human inheritance's property cardiomyopathy of the invention has and human inheritance's property heart The similar disease phenotype of patients with myopathy cardiac muscle cell and electrophysiological change, derive from a wealth of sources, and can long-term in vitro culture.
In another example Chinese patent CN201610005683.9 is disclosed and a kind of being carried out in-situ immobilization to coagulation factor F8/F9 Method, the mutational site for selecting coagulation factor gene is the gene loci of in-situ immobilization, design CRISPR/Cas systems Homologous recombination donor sequences are inserted into gene in-situ immobilization site by the binding site of the nuclease of sgRNA sequences, repair base The expression of cause or supplement gene, the invention method specify research method for the in-situ immobilization in coagulation factor mutational site and face The possibility of bed application.
Again for example, Chinese patent CN105985985A removes allosome mescenchymal stem cell table using CRISPR/Cas9 technologies Face leads to the antigen of immunological rejection and causes the inflammatory factor of inflammatory reaction, and improves the anti-apoptotic energy of mescenchymal stem cell Power promotes it to go back to the nest, and the preparation for the drug of clinical treatment angiocardiopathy provides a set of new technical solution, and prepared is different It will not cause immunological rejection after body mesenchymal stem cell transplantation.
Although CRISPR/Cas9 systems have many other application examples at present, however there is no utilize CRISPR/ at present Cas9 systems repair the application of IL-2RG defects, this is because all multivariables of carrier sequence, specific experiment parameter all can be to repairing There is influence in multiple effect, at present also without any technological guidance for solving these problems.
Invention content
To achieve the above object, CRISPR/Cas9 system homologous recombination repairs being utilized the present invention provides a kind of
The method of IL-2RG dcc genes, the present invention is by obtaining pAAV-AAVS1 Donor carriers and pAAV-SaCas9- GRNA AAVS1 carriers are for repairing IL-2RG dcc genes.By by its co-implanted host cell, pAAV-SaCas9- CRISPR/Cas9 systems in gRNA AAVS1 carriers cut defect IL-2RG genes in host cell, and using same Correct IL-2RG genes in pAAV-AAVS1 Donor carriers are properly inserted in genome by source recombinantal repair mechanism.
The invention is realized by the following technical scheme：
On the one hand, CRISPR/Cas9 system homologous recombination repair IL-2RG dcc genes being utilized the present invention provides a kind of System, the system includes two carriers, contains CRISPR/Cas9 systems on one of carrier, on another carrier Contain correct IL-2RG genes；The carrier containing CRISPR/Cas9 systems is named as pAAV-SaCas9-gRNA AAVS1 carriers, nucleotide sequence is as shown in SEQ ID No.1；The carrier containing correct IL-2RG genes is named as PAAV-AAVS1 Donor carriers, nucleotide sequence is as shown in SEQ ID No.2.
PAAV-SaCas9-gRNA AAVS1 carriers in above system cut place for providing CRISPR/Cas9 systems The defects of chief cell IL-2RG genes；PAAV-AAVS1 Donor carriers provide correct IL-2RG genes as donor, adopt It is properly inserted in genome with homologous recombination repair mechanism.
On the other hand, CRISPR/Cas9 system homologous recombination repair IL-2RG defect bases being utilized the present invention provides a kind of The method of cause, the method are：Using the pAAV-SaCas9-gRNA AAVS1 carriers containing CRISPR/Cas9 systems and contain There are the pAAV-AAVS1 Donor carriers of correct IL-2RG genes to repair IL-2RG dcc genes；Wherein pAAV-SaCas9-gRNA The nucleotide sequence of AAVS1 carriers is as shown in SEQ ID No.1, the nucleotide sequence such as SEQ of pAAV-AAVS1 Donor carriers Shown in ID No.2.
The method is specially to be cut using the pAAV-SaCas9-gRNA AAVS1 carriers containing CRISPR/Cas9 systems Cut the defects of host cell IL-2RG genes；To contain the pAAV-AAVS1 Donor carriers of correct IL-2RG genes as supplying Body provides correct IL-2RG genes, is properly inserted in genome using homologous recombination repair mechanism.
More specifically, the method includes the following steps：
(1) pAAV-AAVS1 Donor carriers and pAAV-SaCas9-gRNA AAVS1 carriers are obtained；
(2) that pAAV-SaCas9-gRNA AAVS1 carriers and pAAV-AAVS1 Donor carriers are imported host jointly is thin Born of the same parents.
The method that above-mentioned steps (1) obtain pAAV-AAVS1 Donor carriers can be to be appointed using commonly used in the art Where method obtains the carrier, such as complete sequence synthesis, PCR (PCR) or passes through commercial sources and buy etc..
The method that above-mentioned steps (1) obtain pAAV-SaCas9-gRNA AAVS1 carriers can be routinely to be made using this field Any method obtains the carrier, such as complete sequence synthesis, PCR (PCR) or is bought by commercial sources Deng.
Above-mentioned steps (2) pAAV-SaCas9-gRNA AAVS1 carriers and pAAV-AAVS1 Donor carriers import place jointly Any method commonly used in the art may be used in chief cell.
In another aspect, the present invention provides a kind of kit for repairing IL-2RG dcc genes, the kit Including above-mentioned pAAV-SaCas9-gRNA AAVS1 carriers and pAAV-AAVS1 Donor carriers.
The kit further includes by above-mentioned pAAV-SaCas9-gRNA AAVS1 carriers and pAAV-AAVS1 Donor The required part or all of reagent of vector introduction host cell.
Compared with prior art, beneficial effects of the present invention are：
The method that the present invention utilizes CRISPRCas9 system homologous recombination repair IL-2RG dcc genes, realizes X-SCID The breakthrough precisely treated.
Specific patient X-SCID of diagnosis recruited using clinic, establishes X-SCID humanized mouse models, will be of the invention The pAAV-SaCas9-gRNA AAVS1 carriers and pAAV-AAVS1 Donor carriers of offer import in Mice Body jointly, after 4 weeks, The reaction of IL-2RG gene PCRs and sequencing and immune function assessment are carried out, CD127 is completed, CD132 Protein Detections, are immunized blood routine The inspections such as globulin, Lymphocyte subset, lymphopoiesis, TREC, TCR diversity, testing result show IL-2RG genes Expression is normal.
Specific implementation mode
Below in conjunction with embodiment, further the present invention will be described in detail, it should be pointed out that following explanation is only pair Claimed technical solution for example, not to any restrictions of these technical solutions, protection of the invention Range be subject to the appended claims record content.
Do not make the molecular biology test method illustrated, equal reference in following embodiment《Molecular Cloning:A Laboratory guide》 Listed specific method carries out in one book of (third edition) J. Pehanorm Brookers, or is carried out according to kit and product description； The kit biomaterial commercially obtains unless otherwise specified.Those skilled in the art should understand that It is that can the details and form of technical solution of the present invention be modified or be replaced without departing from the spirit and scope of the invention It changes, but these modifications and replacement are each fallen in protection scope of the present invention.
Embodiment 1：The acquisition of pAAV-AAVS1 Donor carriers
PAAV-AAVS1 Donor carrier construction methods include the following steps：
(1) using human source gene group DNA as template, using LHANotIF, LHA-CAGminiR as primer, PCR obtains LHA sequences Row；LHANotIF, LHA-CAGminiR primer base sequences are as shown in SEQ ID No.3 and SEQ ID No.4；LHA sequences Nucleotide sequence is as shown in SEQ ID No.15；
(2) to tie up true carrier pAV-CAGmini as template, using LHA-CAGmini-F, CAG-NM-000206-R as primer, PCR obtains CAGmini sequences；LHA-CAGmini-F, CAG-NM-000206-R primer base sequences such as SEQ ID No.5 Shown in SEQ ID No.6；The nucleotide sequence of CAGmini sequences is as shown in SEQ ID No.16, the true carrier of the dimension PAV-CAGmini is the carrier that applicant independently builds, and nucleotide sequence is as shown in SEQ ID No.17；
(3) to tie up true carrier pENTER-NM-000206 as template, with CAG-NM-000206-F, NM-000206-P2A- GFP-R is primer, and PCR obtains IL-2RG sequences；CAG-NM-000206-F, NM-000206-P2A-GFP-R primer base Sequence is as shown in SEQ ID No.7 and SEQ ID No.8；The nucleotide sequence of IL-2RG sequences as shown in SEQ ID No.18, The true carrier pENTER-NM-000206 of dimension is the carrier that applicant independently builds, nucleotide sequence such as SEQ ID No.19 It is shown；
(4) to tie up true carrier pAV-P2A-GFP as template, NM-000206-P2A-GFP-F, P2A-GFP-SV40-R are to draw Object, PCR obtain P2A-GFP sequences, NM-000206-P2A-GFP-F, P2A-GFP-SV40-R primer base sequences such as SEQ Shown in ID No.9 and SEQ ID No.10；As shown in SEQ.No.20, the dimension really carries the nucleotide sequence of P2A-GFP sequences Body pAV-P2A-GFP is the carrier that applicant independently builds, and nucleotide sequence is as shown in SEQ ID No.21；
(5) to tie up true carrier pENTER as template, P2A-GFP-SV40-F, SV40-RHA-R are primer, and PCR obtains SV40 Sequence, P2A-GFP-SV40-F, SV40-RHA-R primer base sequences such as SEQ ID No.11 and SEQ ID No.12 institutes Show；For the nucleotide sequence of SV40 sequences as shown in SEQ.No.22, the true carrier pENTER of dimension is what applicant independently built Carrier, nucleotide sequence is as shown in SEQ ID No.23；
(6) using human source gene group DNA as template, SV40-RHA-F, RHA-NotI-R are primer, and PCR obtains RHA sequences, SV40-RHA-F, RHA-NotI-R primer base sequences are as shown in SEQ ID No.13 and SEQ ID No.14；RHA sequences Nucleotide sequence as shown in SEQ.No.24；
(7) obtained with above-mentioned steps (1)-(6) six sections of PCR products (LHA sequences, CAGmini sequences, IL-2RG sequences, P2A-GFP sequences, SV40 sequences and RHA sequences) it is template, using LHA-NotI-F, RHA-NotI-R as primer, PCR obtains LHA- CAGmini-IL2RG-P2A-GFP-RHA full length sequences；The nucleotides sequence of LHA-CAGmini-IL2RG-P2A-GFP-RHA sequences Row are as shown in SEQ.No.25；
(8) the PCR product LHA-CAGmini-IL2RG-P2A- obtained with restriction enzyme NotI single endonuclease digestions step (7) GFP-RHA and the true carrier pAV-U6-GFP carriers of dimension, digestion system are as follows：
The nucleotide sequence of the true carrier pAV-U6-GFP carriers of dimension is as shown in SEQ ID No.26；
(9) digestion products that recycling step (8) obtains obtain the NotI single endonuclease digestion products of pAV-U6-GFP carriers, and The NotI single endonuclease digestion products of LHA-CAGmini-IL2RG-P2A-GFP-RHA, and the two is connected into 2h to get pAAV- in 22 DEG C AAVS1 Donor carriers, linked system are as follows：
(10) connection product for obtaining step (9) converts bacillus coli DH 5 alpha competent cell, is coated on the LB containing AMP Positive-selecting is carried out on tablet, the clone built NotI single endonuclease digestions are verified, and will verify correct Cloning of full length sequencing, i.e., Obtain pAAV-AAVS1 Donor carrier clonings.
The PCR system of above-mentioned steps (1)-(7) is as follows：
The PCR response procedures of above-mentioned steps (1)-(7) are：
Embodiment 2：The acquisition of pAAV-SaCas9-gRNA AAVS1 carriers
The acquisition pattern of pAAV-SaCas9-gRNA AAVS1 carriers is：
The pAAV-SaCas9-gRNA AAVS1 skeleton carriers pAAV-SaCas9-gRNA purchases are in Addgene public affairs Department, product identification 61591.
PAAV-SaCas9-gRNA AAVS1 vector construction processes：
(1) primer AAVS1-gRNA-F and AAVS1-gRNA-R are bought,
The primer AAVS1-gRNA-F, base sequence are CACCGAGAGCCACATTAAC CGGCCCTG；
Described its base sequence of primer AAVS1-gRNA-R is AAACCAGGGCCGGTTAAT GTGGC TCTC.
(2) use restriction enzyme BsaI single endonuclease digestion skeleton carrier pAAV-SaCas9-gRNA, digestion system as follows：
(3) it anneals, receives first brief centrifugation after the primer in step (1), be then diluted to 100 μM of mother liquor, wherein Buffer 3.1 is purchased from NEB companies, and article No. is NEB B7203S.
The annealing reaction system is as follows：
The response procedures are as follows：
(4) digestion products that recycling step (2) obtains obtain the BsaI single endonuclease digestions production of pAAV-SaCas9-gRNA carriers Annealed product is diluted 100 times by object, and 2h is connect in 22 DEG C with the good carrier of digestion to get to pAAV-SaCas9-gRNA AAVS1 carriers, linked system are as follows：
(5) connection product is converted into bacillus coli DH 5 alpha competent cell, is coated on the LB tablets containing AMP and carries out the positive Screening, sequence verification is to get to pAAV-SaCas9-gRNA AAVS1 carriers.
Embodiment 3：Utilize the zoopery of CRISPR/Cas9 system homologous recombination repair IL-2RG dcc genes
(1) X-SCID humanized mouse models are built：
It is engaged in primary immunodeficiency disease clinic and research work unit X linked severe combined immunodeficiency diseases using the whole nation (X-SCID) clinical resources recruit specific patient X-SCID of gene diagnosis.Establish X-SCID humanized mouse models：Acquisition Candidate stem cell after magnetic bead sorting CD34+ candidate stem cells, is squeezed into NSG mouse after 4 weeks and adopted by patient X-SCID marrow 2mL Collect the mosaic status of a little peripheral blood detection humanizing cells；It is small to get X-SCID humanizations after the success of humanization mouse chimera Mouse model；
(2) will the 1 obtained homologous recombination donor pAAV-AAVS1 containing IL-2RG correct genes be implemented by the present invention The pAAV-SaCas9-gRNA AAVS1 containing CRISPR/Cas9 systems that Donor carriers and the embodiment of the present invention 2 obtain are carried The complex carries system of body composition, tail vein injection enter in Mice Body.Human archeocyte infection conditions are detected after 2 weeks, by people source after 4 weeks Lymphocyte sorting is T cell, B cell, NK cells, granulocyte, monocyte, after extracting DNA, carries out IL-2RG gene PCRs Reaction and sequencing and immune function assessment, complete CD127, CD132 Protein Detections, blood routine, immunoglobulin, lymphocyte point The inspections such as class, lymphopoiesis, TREC, TCR diversity.Testing result shows that IL-2RG gene expressions are normal.
More than, it is illustrated based on embodiments of the present invention, however, the present invention is not limited thereto, those skilled in the art exist Modifications and changes embodiment is carried out in the main scope of the present invention, should all be belonged to the scope of protection of the present invention.
Sequence table
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cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt 60
ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 120
aggggttcct gcggcctcta gactcgaggc gttgacattg attattgact agttattaat 180
agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac 240
ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa 300
tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt 360
atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc 420
ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat 480
gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc 540
ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc 600
tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa 660
aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg 720
tctatataag cagagctctc tggctaacta ccggtgccac catggcccca aagaagaagc 780
ggaaggtcgg tatccacgga gtcccagcag ccaagcggaa ctacatcctg ggcctggaca 840
tcggcatcac cagcgtgggc tacggcatca tcgactacga gacacgggac gtgatcgatg 900
ccggcgtgcg gctgttcaaa gaggccaacg tggaaaacaa cgagggcagg cggagcaaga 960
gaggcgccag aaggctgaag cggcggaggc ggcatagaat ccagagagtg aagaagctgc 1020
tgttcgacta caacctgctg accgaccaca gcgagctgag cggcatcaac ccctacgagg 1080
ccagagtgaa gggcctgagc cagaagctga gcgaggaaga gttctctgcc gccctgctgc 1140
acctggccaa gagaagaggc gtgcacaacg tgaacgaggt ggaagaggac accggcaacg 1200
agctgtccac caaagagcag atcagccgga acagcaaggc cctggaagag aaatacgtgg 1260
ccgaactgca gctggaacgg ctgaagaaag acggcgaagt gcggggcagc atcaacagat 1320
tcaagaccag cgactacgtg aaagaagcca aacagctgct gaaggtgcag aaggcctacc 1380
accagctgga ccagagcttc atcgacacct acatcgacct gctggaaacc cggcggacct 1440
actatgaggg acctggcgag ggcagcccct tcggctggaa ggacatcaaa gaatggtacg 1500
agatgctgat gggccactgc acctacttcc ccgaggaact gcggagcgtg aagtacgcct 1560
acaacgccga cctgtacaac gccctgaacg acctgaacaa tctcgtgatc accagggacg 1620
agaacgagaa gctggaatat tacgagaagt tccagatcat cgagaacgtg ttcaagcaga 1680
agaagaagcc caccctgaag cagatcgcca aagaaatcct cgtgaacgaa gaggatatta 1740
agggctacag agtgaccagc accggcaagc ccgagttcac caacctgaag gtgtaccacg 1800
acatcaagga cattaccgcc cggaaagaga ttattgagaa cgccgagctg ctggatcaga 1860
ttgccaagat cctgaccatc taccagagca gcgaggacat ccaggaagaa ctgaccaatc 1920
tgaactccga gctgacccag gaagagatcg agcagatctc taatctgaag ggctataccg 1980
gcacccacaa cctgagcctg aaggccatca acctgatcct ggacgagctg tggcacacca 2040
acgacaacca gatcgctatc ttcaaccggc tgaagctggt gcccaagaag gtggacctgt 2100
cccagcagaa agagatcccc accaccctgg tggacgactt catcctgagc cccgtcgtga 2160
agagaagctt catccagagc atcaaagtga tcaacgccat catcaagaag tacggcctgc 2220
ccaacgacat cattatcgag ctggcccgcg agaagaactc caaggacgcc cagaaaatga 2280
tcaacgagat gcagaagcgg aaccggcaga ccaacgagcg gatcgaggaa atcatccgga 2340
ccaccggcaa agagaacgcc aagtacctga tcgagaagat caagctgcac gacatgcagg 2400
aaggcaagtg cctgtacagc ctggaagcca tccctctgga agatctgctg aacaacccct 2460
tcaactatga ggtggaccac atcatcccca gaagcgtgtc cttcgacaac agcttcaaca 2520
acaaggtgct cgtgaagcag gaagaaaaca gcaagaaggg caaccggacc ccattccagt 2580
acctgagcag cagcgacagc aagatcagct acgaaacctt caagaagcac atcctgaatc 2640
tggccaaggg caagggcaga atcagcaaga ccaagaaaga gtatctgctg gaagaacggg 2700
acatcaacag gttctccgtg cagaaagact tcatcaaccg gaacctggtg gataccagat 2760
acgccaccag aggcctgatg aacctgctgc ggagctactt cagagtgaac aacctggacg 2820
tgaaagtgaa gtccatcaat ggcggcttca ccagctttct gcggcggaag tggaagttta 2880
agaaagagcg gaacaagggg tacaagcacc acgccgagga cgccctgatc attgccaacg 2940
ccgatttcat cttcaaagag tggaagaaac tggacaaggc caaaaaagtg atggaaaacc 3000
agatgttcga ggaaaagcag gccgagagca tgcccgagat cgaaaccgag caggagtaca 3060
aagagatctt catcaccccc caccagatca agcacattaa ggacttcaag gactacaagt 3120
acagccaccg ggtggacaag aagcctaata gagagctgat taacgacacc ctgtactcca 3180
cccggaagga cgacaagggc aacaccctga tcgtgaacaa tctgaacggc ctgtacgaca 3240
aggacaatga caagctgaaa aagctgatca acaagagccc cgaaaagctg ctgatgtacc 3300
accacgaccc ccagacctac cagaaactga agctgattat ggaacagtac ggcgacgaga 3360
agaatcccct gtacaagtac tacgaggaaa ccgggaacta cctgaccaag tactccaaaa 3420
aggacaacgg ccccgtgatc aagaagatta agtattacgg caacaaactg aacgcccatc 3480
tggacatcac cgacgactac cccaacagca gaaacaaggt cgtgaagctg tccctgaagc 3540
cctacagatt cgacgtgtac ctggacaatg gcgtgtacaa gttcgtgacc gtgaagaatc 3600
tggatgtgat caaaaaagaa aactactacg aagtgaatag caagtgctat gaggaagcta 3660
agaagctgaa gaagatcagc aaccaggccg agtttatcgc ctccttctac aacaacgatc 3720
tgatcaagat caacggcgag ctgtatagag tgatcggcgt gaacaacgac ctgctgaacc 3780
ggatcgaagt gaacatgatc gacatcacct accgcgagta cctggaaaac atgaacgaca 3840
agaggccccc caggatcatt aagacaatcg cctccaagac ccagagcatt aagaagtaca 3900
gcacagacat tctgggcaac ctgtatgaag tgaaatctaa gaagcaccct cagatcatca 3960
aaaagggcaa aaggccggcg gccacgaaaa aggccggcca ggcaaaaaag aaaaagggat 4020
cctacccata cgatgttcca gattacgctt acccatacga tgttccagat tacgcttacc 4080
catacgatgt tccagattac gcttaagaat tcctagagct cgctgatcag cctcgactgt 4140
gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga 4200
aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag 4260
taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga 4320
agagaatagc aggcatgctg gggaggtacc gagggcctat ttcccatgat tccttcatat 4380
ttgcatatac gatacaaggc tgttagagag ataattggaa ttaatttgac tgtaaacaca 4440
aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt 4500
ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat 4560
ttcttggctt tatatatctt gtggaaagga cgaaacaggg ccggttaatg tggctctcgg 4620
tggttttagt actctggaaa cagaatctac taaaacaagg caaaatgccg tgtttatctc 4680
gtcaacttgt tggcgagatt tttgcggccg caggaacccc tagtgatgga gttggccact 4740
ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg 4800
ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg 4860
atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac 4920
catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg 4980
tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc 5040
tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc 5100
gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta 5160
gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta 5220
atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg 5280
atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa 5340
aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta 5400
caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg 5460
cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg 5520
ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc 5580
tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag 5640
gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt 5700
caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa 5760
ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt 5820
gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt 5880
tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt 5940
ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg 6000
tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga 6060
atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa 6120
gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga 6180
caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa 6240
ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca 6300
ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta 6360
ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac 6420
ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc 6480
gtggaagccg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag 6540
ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga 6600
taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt 6660
agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata 6720
atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag 6780
aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa 6840
caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt 6900
ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc 6960
cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa 7020
tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa 7080
gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc 7140
ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa 7200
gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa 7260
caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg 7320
ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc 7380
tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg 7440
ctcacatgt 7449
<210> 2
<211> 7106
<212> DNA
<213>Artificial sequence (RenGongXuLie)
<400> 2
cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt 60
ggtcgcccgg cctcagtgag cgagcgcgca gagagggagt ggccaactcc atcactaggg 120
gttcctgcgg ccgccatgca tcactttgag ctctactggc ttctgcgccg cctctggccc 180
actgtttccc cttcccaggc aggtcctgct ttctctgacc tgcattctct cccctgggcc 240
tgtgccgctt tctgtctgca gcttgtggcc tgggtcacct ctacggctgg cccagatcct 300
tccctgccgc ctccttcagg ttccgtcttc ctccactccc tcttcccctt gctctctgct 360
gtgttgctgc ccaaggatgc tctttccgga gcacttcctt ctcggcgctg caccacgtga 420
tgtcctctga gcggatcctc cccgtgtctg ggtcctctcc gggcatctct cctccctcac 480
ccaaccccat gccgtcttca ctcgctgggt tcccttttcc ttctccttct ggggcctgtg 540
ccatctctcg tttcttagga tggccttctc cgacggatgt ctcccttgcg tcccgcctcc 600
ccttcttgta ggcctgcatc atcaccgttt ttctggacaa ccccaaagta ccccgtctcc 660
ctggctttag ccacctctcc atcctcttgc tttctttgcc tggacacccc gttctcctgt 720
ggattcgggt cacctctcac tcctttcatt tgggcagctc ccctaccccc cttacctctc 780
tagtctgtgc tagctcttcc agccccctgt catggcatct tccaggggtc cgagagctca 840
gctagtcttc ttcctccaac ccgggcccct atgtccactt caggacagca tgtttgctgc 900
ctccagggat cctgtgtccc cgagctggga ccaccttata ttcccagggc cggttaacta 960
gtacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga 1020
cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat 1080
gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa 1140
gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca 1200
tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc aagcttctcg 1260
agggcgcgtg acctcaccct gtggagccac accctagggt tggccaatct actcccagga 1320
gcagggaggg caggagccag ggctgggcat aaaagtcagg gcagagccat ctattgcttg 1380
cgatcgccac catgttgaag ccatcattac cattcacatc cctcttattc ctgcagctgc 1440
ccctgctggg agtggggctg aacacgacaa ttctgacgcc caatgggaat gaagacacca 1500
cagctgattt cttcctgacc actatgccca ctgactccct cagtgtttcc actctgcccc 1560
tcccagaggt tcagtgtttt gtgttcaatg tcgagtacat gaattgcact tggaacagca 1620
gctctgagcc ccagcctacc aacctcactc tgcattattg gtacaagaac tcggataatg 1680
ataaagtcca gaagtgcagc cactatctat tctctgaaga aatcacttct ggctgtcagt 1740
tgcaaaaaaa ggagatccac ctctaccaaa catttgttgt tcagctccag gacccacggg 1800
aacccaggag acaggccaca cagatgctaa aactgcagaa tctggtgatc ccctgggctc 1860
cagagaacct aacacttcac aaactgagtg aatcccagct agaactgaac tggaacaaca 1920
gattcttgaa ccactgtttg gagcacttgg tgcagtaccg gactgactgg gaccacagct 1980
ggactgaaca atcagtggat tatagacata agttctcctt gcctagtgtg gatgggcaga 2040
aacgctacac gtttcgtgtt cggagccgct ttaacccact ctgtggaagt gctcagcatt 2100
ggagtgaatg gagccaccca atccactggg ggagcaatac ttcaaaagag aatcctttcc 2160
tgtttgcatt ggaagccgtg gttatctctg ttggctccat gggattgatt atcagccttc 2220
tctgtgtgta tttctggctg gaacggacga tgccccgaat tcccaccctg aagaacctag 2280
aggatcttgt tactgaatac cacgggaact tttcggcctg gagtggtgtg tctaagggac 2340
tggctgagag tctgcagcca gactacagtg aacgactctg cctcgtcagt gagattcccc 2400
caaaaggagg ggcccttggg gaggggcctg gggcctcccc atgcaaccag catagcccct 2460
actgggcccc cccatgttac accctaaagc ctgaaaccgg aagcggagct actaacttca 2520
gcctgctgaa gcaggctgga gacgtggagg agaaccctgg acctgtcgac atggtgagca 2580
agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa 2640
acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga 2700
ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca 2760
ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact 2820
tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg 2880
acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca 2940
tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt 3000
acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg 3060
tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc 3120
agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca 3180
cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt 3240
tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa acgcgtgatc 3300
caccggatct agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc 3360
tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt 3420
tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt 3480
cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt 3540
atcttaacag atctatgtgg ctctggttct gggtactttt atctgtcccc tccaccccac 3600
agtggggcca ctagggacag gattggtgac agaaaagccc catccttagg cctcctcctt 3660
cctagtctcc tgatattggg tctaaccccc acctcctgtt aggcagattc cttatctggt 3720
gacacacccc catttcctgg agccatctct ctccttgcca gaacctctaa ggtttgctta 3780
cgatggagcc agagaggatc ctgggaggga gagcttggca gggggtggga gggaaggggg 3840
ggatgcgtga cctgcccggt tctcagtggc caccctgcgc taccctctcc cagaacctga 3900
gctgctctga cgcggccgtc tggtgcgttt cactgatcct ggtgctgcag cttccttaca 3960
cttcccaaga ggagaagcag tttggaaaaa caaaatcaga ataagttggt cctgagttct 4020
aactttggct cttcaccttt ctagtcccca atttatattg ttcctccgtg cgtcagtttt 4080
acctgtgaga taaggccagt agccagcccc gtcctggcag ggctgtggtg aggagggggg 4140
tgtccgtgtg gaaaactccc tttgtgagaa tggtgcgtcc taggtgttca ccaggtcgtg 4200
gccgcctcta ctccctttct ctttctccat ccttctttcc ttaaagagtc cccagtgcta 4260
tctgggacat attcctccgc ccagagcagg gtcccgcttc cctaaggccc tgctctgggc 4320
ttctgggttt gagtccttgg caagcccagg agagtctaga gcggccgcag gaacccctag 4380
tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa 4440
aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct 4500
gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc 4560
gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt 4620
ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc 4680
tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg 4740
gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt 4800
gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt 4860
ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat 4920
ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa 4980
tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt 5040
atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc 5100
gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca 5160
agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg 5220
cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat 5280
ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt 5340
atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct 5400
tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc 5460
cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa 5520
agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg 5580
taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt 5640
tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg 5700
catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac 5760
ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc 5820
ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa 5880
catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc 5940
aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt 6000
aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga 6060
taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa 6120
atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa 6180
gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa 6240
tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt 6300
ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt 6360
gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg 6420
agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt 6480
aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca 6540
agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac 6600
tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac 6660
atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct 6720
taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg 6780
gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca 6840
gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt 6900
aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta 6960
tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc 7020
gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc 7080
cttttgctgg ccttttgctc acatgt 7106
<210> 3
<211> 41
<212> DNA
<213>Artificial sequence (RenGongXuLie)
<400> 3
gcgtgcggcc gccatgcatc actttgagct ctactggctt c 41
<210> 4
<211> 49
<212> DNA
<213>Artificial sequence (RenGongXuLie)
<400> 4
gccatttacc gtaagttatg tactagttaa ccggccctgg gaatataag 49
<210> 5
<211> 49
<212> DNA
<213>Artificial sequence (RenGongXuLie)
<400> 5
cttatattcc cagggccggt taactagtac ataacttacg gtaaatggc 49
<210> 6
<211> 52
<212> DNA
<213>Artificial sequence (RenGongXuLie)
<400> 6
ggtaatgatg gcttcaacat ggtggcgatc gcaagcaata gatggctctg cc 52
<210> 7
<211> 52
<212> DNA
<213>Artificial sequence (RenGongXuLie)
<400> 7
ggcagagcca tctattgctt gcgatcgcca ccatgttgaa gccatcatta cc 52
<210> 8
<211> 40
<212> DNA
<213>Artificial sequence (RenGongXuLie)
<400> 8
gaagttagta gctccgcttc cggtttcagg ctttagggtg 40
<210> 9
<211> 37
<212> DNA
<213>Artificial sequence (RenGongXuLie)
<400> 9
caccctaaag cctgaaaccg gaagcggagc tactaac 37
<210> 10
<211> 46
<212> DNA
<213>Artificial sequence (RenGongXuLie)
<400> 10
gttatctaga tccggtggat cacgcgttta cttgtacagc tcgtcc 46
<210> 11
<211> 46
<212> DNA
<213>Artificial sequence (RenGongXuLie)
<400> 11
ggacgagctg tacaagtaaa cgcgtgatcc accggatcta gataac 46
<210> 12
<211> 48
<212> DNA
<213>Artificial sequence (RenGongXuLie)
<400> 12
cccagaacca gagccacata gatctgttaa gatacattga tgagtttg 48
<210> 13
<211> 48
<212> DNA
<213>Artificial sequence (RenGongXuLie)
<400> 13
caaactcatc aatgtatctt aacagatcta tgtggctctg gttctggg 48
<210> 14
<211> 38
<212> DNA
<213>Artificial sequence (RenGongXuLie)
<400> 14
gcgtgcggcc gctctagact ctcctgggct tgccaagg 38
<210> 15
<211> 815
<212> DNA
<213>Artificial sequence (RenGongXuLie)
<400> 15
cactttgagc tctactggct tctgcgccgc ctctggccca ctgtttcccc ttcccaggca 60
ggtcctgctt tctctgacct gcattctctc ccctgggcct gtgccgcttt ctgtctgcag 120
cttgtggcct gggtcacctc tacggctggc ccagatcctt ccctgccgcc tccttcaggt 180
tccgtcttcc tccactccct cttccccttg ctctctgctg tgttgctgcc caaggatgct 240
ctttccggag cacttccttc tcggcgctgc accacgtgat gtcctctgag cggatcctcc 300
ccgtgtctgg gtcctctccg ggcatctctc ctccctcacc caaccccatg ccgtcttcac 360
tcgctgggtt cccttttcct tctccttctg gggcctgtgc catctctcgt ttcttaggat 420
ggccttctcc gacggatgtc tcccttgcgt cccgcctccc cttcttgtag gcctgcatca 480
tcaccgtttt tctggacaac cccaaagtac cccgtctccc tggctttagc cacctctcca 540
tcctcttgct ttctttgcct ggacaccccg ttctcctgtg gattcgggtc acctctcact 600
cctttcattt gggcagctcc cctacccccc ttacctctct agtctgtgct agctcttcca 660
gccccctgtc atggcatctt ccaggggtcc gagagctcag ctagtcttct tcctccaacc 720
cgggccccta tgtccacttc aggacagcat gtttgctgcc tccagggatc ctgtgtcccc 780
gagctgggac caccttatat tcccagggcc ggtta 815
<210> 16
<211> 417
<212> DNA
<213>Artificial sequence (RenGongXuLie)
<400> 16
acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg 60
tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg 120
gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt 180
acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg 240
accttatggg actttcctac ttggcagtac atctacgtat tagtcatcaa gcttctcgag 300
ggcgcgtgac ctcaccctgt ggagccacac cctagggttg gccaatctac tcccaggagc 360
agggagggca ggagccaggg ctgggcataa aagtcagggc agagccatct attgctt 417
<210> 17
<211> 3737
<212> DNA
<213>Artificial sequence (RenGongXuLie)
<400> 17
cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt 60
ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 120
aggggttcct gcggccggtc gcgtctagta ctagtacata acttacggta aatggcccgc 180
ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag 240
taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc 300
acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg 360
gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc 420
agtacatcta cgtattagtc atcaagcttc tcgagggcgc gtgacctcac cctgtggagc 480
cacaccctag ggttggccaa tctactccca ggagcaggga gggcaggagc cagggctggg 540
cataaaagtc agggcagagc catctattgc ttggtaccga ggagatctgc cgccgcgatc 600
gcyacgcgta cgcggccgct cgaggattat aaggatgacg acgataaatt cgtcgagcac 660
caccaccacc accactaata aggtttatcc gatccaccgg atctagataa gatatccgat 720
ccaccggatc tagataactg atcataatca gccataccac atttgtagag gttttacttg 780
ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg 840
ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt 900
tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg 960
tatcttaacg cggtaaccac gtgcggaccc aacggccgca ggaaccccta gtgatggagt 1020
tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc 1080
gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg 1140
ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc 1200
aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac 1260
gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc 1320
ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt 1380
agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg 1440
ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac 1500
gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcgggcta 1560
ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat 1620
ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac 1680
tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc 1740
cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac 1800
cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg 1860
aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta 1920
gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta 1980
aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata 2040
ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc 2100
ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga 2160
agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct 2220
tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg 2280
tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta 2340
ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat 2400
gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt 2460
acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga 2520
tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga 2580
gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga 2640
actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc 2700
aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc 2760
cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg 2820
tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat 2880
cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata 2940
tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct 3000
ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga 3060
ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg 3120
cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc 3180
aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct 3240
agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc 3300
tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt 3360
ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg 3420
cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct 3480
atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag 3540
ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag 3600
tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg 3660
gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg 3720
gccttttgct cacatgt 3737
<210> 18
<211> 1107
<212> DNA
<213>Artificial sequence (RenGongXuLie)
<400> 18
atgttgaagc catcattacc attcacatcc ctcttattcc tgcagctgcc cctgctggga 60
gtggggctga acacgacaat tctgacgccc aatgggaatg aagacaccac agctgatttc 120
ttcctgacca ctatgcccac tgactccctc agtgtttcca ctctgcccct cccagaggtt 180
cagtgttttg tgttcaatgt cgagtacatg aattgcactt ggaacagcag ctctgagccc 240
cagcctacca acctcactct gcattattgg tacaagaact cggataatga taaagtccag 300
aagtgcagcc actatctatt ctctgaagaa atcacttctg gctgtcagtt gcaaaaaaag 360
gagatccacc tctaccaaac atttgttgtt cagctccagg acccacggga acccaggaga 420
caggccacac agatgctaaa actgcagaat ctggtgatcc cctgggctcc agagaaccta 480
acacttcaca aactgagtga atcccagcta gaactgaact ggaacaacag attcttgaac 540
cactgtttgg agcacttggt gcagtaccgg actgactggg accacagctg gactgaacaa 600
tcagtggatt atagacataa gttctccttg cctagtgtgg atgggcagaa acgctacacg 660
tttcgtgttc ggagccgctt taacccactc tgtggaagtg ctcagcattg gagtgaatgg 720
agccacccaa tccactgggg gagcaatact tcaaaagaga atcctttcct gtttgcattg 780
gaagccgtgg ttatctctgt tggctccatg ggattgatta tcagccttct ctgtgtgtat 840
ttctggctgg aacggacgat gccccgaatt cccaccctga agaacctaga ggatcttgtt 900
actgaatacc acgggaactt ttcggcctgg agtggtgtgt ctaagggact ggctgagagt 960
ctgcagccag actacagtga acgactctgc ctcgtcagtg agattccccc aaaaggaggg 1020
gcccttgggg aggggcctgg ggcctcccca tgcaaccagc atagccccta ctgggccccc 1080
ccatgttaca ccctaaagcc tgaaacc 1107
<210> 19
<211> 8578
<212> DNA
<213>Artificial sequence (RenGongXuLie)
<400> 19
ttcgaattta aattaattaa catcatcaat aatatacctt attttggatt gaagccaata 60
tgataatgag ggggtggagt ttgtgacgtg gcgcggggcg tgggaacggg gcgggtgacg 120
tagtagtgtg gcggaagtgt gatgttgcaa gtgtggcgga acacatgtaa gcgacggatg 180
tggcaaaagt gacgtttttg gtgtgcgccg gtgtacacag gaagtgacaa ttttcgcgcg 240
gttttaggcg gatgttgtag taaatttggg cgtaaccgag taagatttgg ccattttcgc 300
gggaaaactg aataagagga agtgaaatct gaataatttt gtgttactca tagcgcgtaa 360
tactgtaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg 420
ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga 480
cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat 540
gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa 600
gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca 660
tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca 720
tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat 780
ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg 840
actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac 900
ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagtaatacg 960
actcactata gggagaggat ccggtaccga ggagatctgc cgccgcgatc gccatgttga 1020
agccatcatt accattcaca tccctcttat tcctgcagct gcccctgctg ggagtggggc 1080
tgaacacgac aattctgacg cccaatggga atgaagacac cacagctgat ttcttcctga 1140
ccactatgcc cactgactcc ctcagtgttt ccactctgcc cctcccagag gttcagtgtt 1200
ttgtgttcaa tgtcgagtac atgaattgca cttggaacag cagctctgag ccccagccta 1260
ccaacctcac tctgcattat tggtacaaga actcggataa tgataaagtc cagaagtgca 1320
gccactatct attctctgaa gaaatcactt ctggctgtca gttgcaaaaa aaggagatcc 1380
acctctacca aacatttgtt gttcagctcc aggacccacg ggaacccagg agacaggcca 1440
cacagatgct aaaactgcag aatctggtga tcccctgggc tccagagaac ctaacacttc 1500
acaaactgag tgaatcccag ctagaactga actggaacaa cagattcttg aaccactgtt 1560
tggagcactt ggtgcagtac cggactgact gggaccacag ctggactgaa caatcagtgg 1620
attatagaca taagttctcc ttgcctagtg tggatgggca gaaacgctac acgtttcgtg 1680
ttcggagccg ctttaaccca ctctgtggaa gtgctcagca ttggagtgaa tggagccacc 1740
caatccactg ggggagcaat acttcaaaag agaatccttt cctgtttgca ttggaagccg 1800
tggttatctc tgttggctcc atgggattga ttatcagcct tctctgtgtg tatttctggc 1860
tggaacggac gatgccccga attcccaccc tgaagaacct agaggatctt gttactgaat 1920
accacgggaa cttttcggcc tggagtggtg tgtctaaggg actggctgag agtctgcagc 1980
cagactacag tgaacgactc tgcctcgtca gtgagattcc cccaaaagga ggggcccttg 2040
gggaggggcc tggggcctcc ccatgcaacc agcatagccc ctactgggcc cccccatgtt 2100
acaccctaaa gcctgaaacc acgcgtacgc ggccgctcga ggattataag gatgacgacg 2160
ataaattcgt cgagcaccac caccaccacc actaataagg tttatccgat ccaccggatc 2220
tagataagat atccgatcca ccggatctag ataactgatc ataatcagcc ataccacatt 2280
tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa 2340
aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag 2400
caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt 2460
gtccaaactc atcaatgtat cttaacgcgg atctgggcgt ggttaagggt gggaaagaat 2520
atataaggtg ggggtcttat gtagttttgt atctgttttg cagcagccgc cgccgccatg 2580
agcaccaact cgtttgatgg aagcattgtg agctcatatt tgacaacgcg catgccccca 2640
tgggccgggg tgcgtcagaa tgtgatgggc tccagcattg atggtcgccc cgtcctgccc 2700
gcaaactcta ctaccttgac ctacgagacc gtgtctggaa cgccgttgga gactgcagcc 2760
tccgccgccg cttcagccgc tgcagccacc gcccgcggga ttgtgactga ctttgctttc 2820
ctgagcccgc ttgcaagcag tgcagcttcc cgttcatccg cccgcgatga caagttgacg 2880
gctcttttgg cacaattgga ttctttgacc cgggaactta atgtcgtttc tcagcagctg 2940
ttggatctgc gccagcaggt ttctgccctg aaggcttcct cccctcccaa tgcggtttaa 3000
aacataaata aaaaaccaga ctctgtttgg atttggatca agcaagtgtc ttgctgtctt 3060
tatttagggg ttttgcgcgc gcggtaggcc cgggaccagc ggtctcggtc gttgagggtc 3120
ctgtgtattt tttccaggac gtggtaaagg tgactctgga tgttcagata catgggcata 3180
agcccgtctc tggggtggag gtagcaccac tgcagagctt catgctgcgg ggtggtgttg 3240
tagatgatcc agtcgtagca ggagcgctgg gcgtggtgcc taaaaatgtc tttcagtagc 3300
aagctgattg ccaggggcag gcccttggtg taagtgttta caaagcggtt aagctgggat 3360
gggtgcatac gtgtttaaac tagggataac agggtaatcg gctgtggaat gtgtgtcagt 3420
tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 3480
attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 3540
gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc 3600
taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg 3660
cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg 3720
gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc 3780
aagagacacg tacgaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg 3840
acgtcccccg ggcagtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc 3900
acaccgtcga tccagaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca 3960
cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg 4020
tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca 4080
tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc 4140
cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc 4200
agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg 4260
gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg 4320
gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc 4380
gcaagcccgg tgcctgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc 4440
cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga 4500
aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga 4560
cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat 4620
ggtagggata acagggtaat gtttaaacta taaaatgcaa ggtgctgctc aaaaaatcag 4680
gcaaagcctc gcgcaaaaaa gaaagcacat cgtagtcatg ctcatgcaga taaaggcagg 4740
taagctccgg aaccaccaca gaaaaagaca ccatttttct ctcaaacatg tctgcgggtt 4800
tctgcataaa cacaaaataa aataacaaaa aaacatttaa acattagaag cctgtcttac 4860
aacaggaaaa acaaccctta taagcataag acggactacg gccatgccgg cgtgaccgta 4920
aaaaaactgg tcaccgtgat taaaaagcac caccgacagc tcctcggtca tgtccggagt 4980
cataatgtaa gactcggtaa acacatcagg ttgattcaca tcggtcagtg ctaaaaagcg 5040
accgaaatag cccgggggaa tacatacccg caggcgtaga gacaacatta cagcccccat 5100
aggaggtata acaaaattaa taggagagaa aaacacataa acacctgaaa aaccctcctg 5160
cctaggcaaa atagcaccct cccgctccag aacaacatac agcgcttcca cagcggcagc 5220
cataacagtc agccttacca gtaaaaaaga aaacctatta aaaaaacacc actcgacacg 5280
gcaccagctc aatcagtcac agtgtaaaaa agggccaagt gcagagcgag tatatatagg 5340
actaaaaaat gacgtaacgg ttaaagtcca caaaaaacac ccagaaaacc gcacgcgaac 5400
ctacgcccag aaacgaaagc caaaaaaccc acaacttcct caaatcgtca cttccgtttt 5460
cccacgttac gtcacttccc attttaagaa aactacaatt cccaacacat acaagttact 5520
ccgccctaaa acctacgtca cccgccccgt tcccacgccc cgcgccacgt cacaaactcc 5580
accccctcat tatcatattg gcttcaatcc aaaataaggt atattattga tgatgttaat 5640
taatttaaat tcgaacatgc atggatccat atgcggtgtg aaataccgca cagatgcgta 5700
aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg 5760
gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca 5820
gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac 5880
cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac 5940
aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg 6000
tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac 6060
ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat 6120
ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag 6180
cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac 6240
ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt 6300
gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt 6360
atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc 6420
aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga 6480
aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac 6540
gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc 6600
cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct 6660
gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca 6720
tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct 6780
ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca 6840
ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc 6900
atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg 6960
cgcaacgttg ttgccattgc tgcagccatg agattatcaa aaaggatctt cacctagatc 7020
cttttcacgt agaaagccag tccgcagaaa cggtgctgac cccggatgaa tgtcagctac 7080
tgggctatct ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg 7140
cttacatggc gatagctaga ctgggcggtt ttatggacag caagcgaacc ggaattgcca 7200
gctggggcgc cctctggtaa ggttgggaag ccctgcaaag taaactggat ggctttcttg 7260
ccgccaagga tctgatggcg caggggatca agctctgatc aagagacagg atgaggatcg 7320
tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg 7380
ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg 7440
ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat 7500
gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca 7560
gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg 7620
gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat 7680
gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa 7740
catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg 7800
gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg 7860
cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg 7920
gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc cgaccgctat 7980
caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac 8040
cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc 8100
cttcttgacg agttcttctg aattttgtta aaatttttgt taaatcagct cattttttaa 8160
ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt 8220
gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa 8280
agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag 8340
ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt 8400
tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg 8460
agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc 8520
cgcgcttaat gcgccgctac agggcgcgtc cattcgccat tcaggatcga attaattc 8578
<210> 20
<211> 792
<212> DNA
<213>Artificial sequence (RenGongXuLie)
<400> 20
ggaagcggag ctactaactt cagcctgctg aagcaggctg gagacgtgga ggagaaccct 60
ggacctgtcg acatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg 120
gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc 180
gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg 240
ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc 300
gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag 360
cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag 420
ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac 480
atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac 540
aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc 600
gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg 660
cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc 720
gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag 780
ctgtacaagt aa 792
<210> 21
<211> 5250
<212> DNA
<213>Artificial sequence (RenGongXuLie)
<400> 21
cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt 60
ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 120
aggggttcct gcggccggtc gcgtctagtt attaatagta atcaattacg gggtcattag 180
ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct 240
gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgt 300
caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg 360
cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat 420
ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca 480
tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc 540
gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga 600
gtttgttttg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt 660
gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt 720
gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg 780
ggaccgatcc agcctccgcg gattcgaatc ccggccggga acggtgcatt ggaacgcgga 840
ttccccgtgc caagagtgac gtaagtaccg cctatagagt ctataggccc acaaaaaatg 900
ctttcttctt ttaatatact tttttgttta tcttatttct aatactttcc ctaatctctt 960
tctttcaggg caataatgat acaatgtatc atgcctcttt gcaccattct aaagaataac 1020
agtgataatt tctgggttaa ggcaatagca atatttctgc atataaatat ttctgcatat 1080
aaattgtaac tgatgtaaga ggtttcatat tgctaatagc agctacaatc cagctaccat 1140
tctgctttta ttttatggtt gggataaggc tggattattc tgagtccaag ctaggccctt 1200
ttgctaatca tgttcatacc tcttatcttc ctcccacagc tcctgggcaa cgtgctggtc 1260
tgtgtgctgg cccatcactt tggcaaagaa ttgggattcg aacatcgatt gaattcagat 1320
ccgctagtaa tacgactcac tatagggaga ggatccggta ccgaggagat ctgccgccgc 1380
gatcgccggc gcgccagatc tcacgcttaa ctagctagcg gaccgacgcg tggaagcgga 1440
gctactaact tcagcctgct gaagcaggct ggagacgtgg aggagaaccc tggacctgtc 1500
gacatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg 1560
gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc 1620
tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc 1680
accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg 1740
aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc 1800
ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc 1860
ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg 1920
cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag 1980
aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc 2040
gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac 2100
cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg 2160
gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag 2220
taagatatcc gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta 2280
gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg 2340
aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat 2400
agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc 2460
aaactcatca atgtatctta acgcggtaac cacgtgcgga cccaacggcc gcaggaaccc 2520
ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga 2580
ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc 2640
agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca 2700
caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg 2760
gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt 2820
tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc 2880
gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg 2940
atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga 3000
cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc 3060
ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa 3120
aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa 3180
ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac 3240
acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca 3300
gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga 3360
aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa 3420
taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt 3480
gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa 3540
tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta 3600
ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag 3660
taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca 3720
gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta 3780
aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc 3840
gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc 3900
ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca 3960
ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc 4020
acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca 4080
taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac 4140
tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg 4200
cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg 4260
ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg 4320
gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac 4380
gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc 4440
aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct 4500
aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc 4560
actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc 4620
gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg 4680
atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa 4740
atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc 4800
ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt 4860
gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa 4920
cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc 4980
tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc 5040
cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct 5100
ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat 5160
gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc 5220
tggccttttg ctggcctttt gctcacatgt 5250
<210> 22
<211> 252
<212> DNA
<213>Artificial sequence (RenGongXuLie)
<400> 22
gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac 60
ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg 120
ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 180
atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 240
atgtatctta ac 252
<210> 23
<211> 7509
<212> DNA
<213>Artificial sequence (RenGongXuLie)
<400> 23
ttcgaattta aattaattaa catcatcaat aatatacctt attttggatt gaagccaata 60
tgataatgag ggggtggagt ttgtgacgtg gcgcggggcg tgggaacggg gcgggtgacg 120
tagtagtgtg gcggaagtgt gatgttgcaa gtgtggcgga acacatgtaa gcgacggatg 180
tggcaaaagt gacgtttttg gtgtgcgccg gtgtacacag gaagtgacaa ttttcgcgcg 240
gttttaggcg gatgttgtag taaatttggg cgtaaccgag taagatttgg ccattttcgc 300
gggaaaactg aataagagga agtgaaatct gaataatttt gtgttactca tagcgcgtaa 360
tactgtaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg 420
ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga 480
cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat 540
gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa 600
gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca 660
tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca 720
tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat 780
ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg 840
actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac 900
ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagtaatacg 960
actcactata gggagaggat ccggtaccga ggagatctgc cgccgcgatc gccggcgcgc 1020
cagatctcaa gcttaactag ctagcggacc gacgcgtacg cggccgctcg aggattataa 1080
ggatgacgac gataaattcg tcgagcacca ccaccaccac cactaataag gtttatccga 1140
tccaccggat ctagataaga tatccgatcc accggatcta gataactgat cataatcagc 1200
cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac 1260
ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt 1320
tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct 1380
agttgtggtt tgtccaaact catcaatgta tcttaacgcg gatctgggcg tggttaaggg 1440
tgggaaagaa tatataaggt gggggtctta tgtagttttg tatctgtttt gcagcagccg 1500
ccgccgccat gagcaccaac tcgtttgatg gaagcattgt gagctcatat ttgacaacgc 1560
gcatgccccc atgggccggg gtgcgtcaga atgtgatggg ctccagcatt gatggtcgcc 1620
ccgtcctgcc cgcaaactct actaccttga cctacgagac cgtgtctgga acgccgttgg 1680
agactgcagc ctccgccgcc gcttcagccg ctgcagccac cgcccgcggg attgtgactg 1740
actttgcttt cctgagcccg cttgcaagca gtgcagcttc ccgttcatcc gcccgcgatg 1800
acaagttgac ggctcttttg gcacaattgg attctttgac ccgggaactt aatgtcgttt 1860
ctcagcagct gttggatctg cgccagcagg tttctgccct gaaggcttcc tcccctccca 1920
atgcggttta aaacataaat aaaaaaccag actctgtttg gatttggatc aagcaagtgt 1980
cttgctgtct ttatttaggg gttttgcgcg cgcggtaggc ccgggaccag cggtctcggt 2040
cgttgagggt cctgtgtatt ttttccagga cgtggtaaag gtgactctgg atgttcagat 2100
acatgggcat aagcccgtct ctggggtgga ggtagcacca ctgcagagct tcatgctgcg 2160
gggtggtgtt gtagatgatc cagtcgtagc aggagcgctg ggcgtggtgc ctaaaaatgt 2220
ctttcagtag caagctgatt gccaggggca ggcccttggt gtaagtgttt acaaagcggt 2280
taagctggga tgggtgcata cgtgtttaaa ctagggataa cagggtaatc ggctgtggaa 2340
tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag 2400
catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag 2460
aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc 2520
catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt 2580
ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg 2640
aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt 2700
cggatctgat caagagacac gtacgaccat gaccgagtac aagcccacgg tgcgcctcgc 2760
cacccgcgac gacgtccccc gggcagtacg caccctcgcc gccgcgttcg ccgactaccc 2820
cgccacgcgc cacaccgtcg atccagaccg ccacatcgag cgggtcaccg agctgcaaga 2880
actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc 2940
cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat 3000
cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg 3060
cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc 3120
gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc 3180
cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta 3240
cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg 3300
gtgcatgacc cgcaagcccg gtgcctgact gtgccttcta gttgccagcc atctgttgtt 3360
tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa 3420
taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg 3480
gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg 3540
gtgggctcta tggtagggat aacagggtaa tgtttaaact ataaaatgca aggtgctgct 3600
caaaaaatca ggcaaagcct cgcgcaaaaa agaaagcaca tcgtagtcat gctcatgcag 3660
ataaaggcag gtaagctccg gaaccaccac agaaaaagac accatttttc tctcaaacat 3720
gtctgcgggt ttctgcataa acacaaaata aaataacaaa aaaacattta aacattagaa 3780
gcctgtctta caacaggaaa aacaaccctt ataagcataa gacggactac ggccatgccg 3840
gcgtgaccgt aaaaaaactg gtcaccgtga ttaaaaagca ccaccgacag ctcctcggtc 3900
atgtccggag tcataatgta agactcggta aacacatcag gttgattcac atcggtcagt 3960
gctaaaaagc gaccgaaata gcccggggga atacataccc gcaggcgtag agacaacatt 4020
acagccccca taggaggtat aacaaaatta ataggagaga aaaacacata aacacctgaa 4080
aaaccctcct gcctaggcaa aatagcaccc tcccgctcca gaacaacata cagcgcttcc 4140
acagcggcag ccataacagt cagccttacc agtaaaaaag aaaacctatt aaaaaaacac 4200
cactcgacac ggcaccagct caatcagtca cagtgtaaaa aagggccaag tgcagagcga 4260
gtatatatag gactaaaaaa tgacgtaacg gttaaagtcc acaaaaaaca cccagaaaac 4320
cgcacgcgaa cctacgccca gaaacgaaag ccaaaaaacc cacaacttcc tcaaatcgtc 4380
acttccgttt tcccacgtta cgtcacttcc cattttaaga aaactacaat tcccaacaca 4440
tacaagttac tccgccctaa aacctacgtc acccgccccg ttcccacgcc ccgcgccacg 4500
tcacaaactc caccccctca ttatcatatt ggcttcaatc caaaataagg tatattattg 4560
atgatgttaa ttaatttaaa ttcgaacatg catggatcca tatgcggtgt gaaataccgc 4620
acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact 4680
cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac 4740
ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa 4800
aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg 4860
acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa 4920
gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc 4980
ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac 5040
gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac 5100
cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg 5160
taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt 5220
atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga 5280
cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct 5340
cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga 5400
ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg 5460
ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct 5520
tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt 5580
aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc 5640
tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg 5700
gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag 5760
atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt 5820
tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag 5880
ttaatagttt gcgcaacgtt gttgccattg ctgcagccat gagattatca aaaaggatct 5940
tcacctagat ccttttcacg tagaaagcca gtccgcagaa acggtgctga ccccggatga 6000
atgtcagcta ctgggctatc tggacaaggg aaaacgcaag cgcaaagaga aagcaggtag 6060
cttgcagtgg gcttacatgg cgatagctag actgggcggt tttatggaca gcaagcgaac 6120
cggaattgcc agctggggcg ccctctggta aggttgggaa gccctgcaaa gtaaactgga 6180
tggctttctt gccgccaagg atctgatggc gcaggggatc aagctctgat caagagacag 6240
gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt 6300
gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg 6360
ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg 6420
gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg 6480
ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg 6540
gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca 6600
tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc 6660
accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc 6720
aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca 6780
aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga 6840
atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg 6900
ccgaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg 6960
aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg 7020
ccttctatcg ccttcttgac gagttcttct gaattttgtt aaaatttttg ttaaatcagc 7080
tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc 7140
gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac 7200
tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca 7260
ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg 7320
agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag 7380
aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc 7440
accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt ccattcgcca ttcaggatcg 7500
aattaattc 7509
<210> 24
<211> 800
<212> DNA
<213>Artificial sequence (RenGongXuLie)
<400> 24
atgtggctct ggttctgggt acttttatct gtcccctcca ccccacagtg gggccactag 60
ggacaggatt ggtgacagaa aagccccatc cttaggcctc ctccttccta gtctcctgat 120
attgggtcta acccccacct cctgttaggc agattcctta tctggtgaca cacccccatt 180
tcctggagcc atctctctcc ttgccagaac ctctaaggtt tgcttacgat ggagccagag 240
aggatcctgg gagggagagc ttggcagggg gtgggaggga agggggggat gcgtgacctg 300
cccggttctc agtggccacc ctgcgctacc ctctcccaga acctgagctg ctctgacgcg 360
gccgtctggt gcgtttcact gatcctggtg ctgcagcttc cttacacttc ccaagaggag 420
aagcagtttg gaaaaacaaa atcagaataa gttggtcctg agttctaact ttggctcttc 480
acctttctag tccccaattt atattgttcc tccgtgcgtc agttttacct gtgagataag 540
gccagtagcc agccccgtcc tggcagggct gtggtgagga ggggggtgtc cgtgtggaaa 600
actccctttg tgagaatggt gcgtcctagg tgttcaccag gtcgtggccg cctctactcc 660
ctttctcttt ctccatcctt ctttccttaa agagtcccca gtgctatctg ggacatattc 720
ctccgcccag agcagggtcc cgcttcccta aggccctgct ctgggcttct gggtttgagt 780
ccttggcaag cccaggagag 800
<210> 25
<211> 4213
<212> DNA
<213>Artificial sequence (RenGongXuLie)
<400> 25
cactttgagc tctactggct tctgcgccgc ctctggccca ctgtttcccc ttcccaggca 60
ggtcctgctt tctctgacct gcattctctc ccctgggcct gtgccgcttt ctgtctgcag 120
cttgtggcct gggtcacctc tacggctggc ccagatcctt ccctgccgcc tccttcaggt 180
tccgtcttcc tccactccct cttccccttg ctctctgctg tgttgctgcc caaggatgct 240
ctttccggag cacttccttc tcggcgctgc accacgtgat gtcctctgag cggatcctcc 300
ccgtgtctgg gtcctctccg ggcatctctc ctccctcacc caaccccatg ccgtcttcac 360
tcgctgggtt cccttttcct tctccttctg gggcctgtgc catctctcgt ttcttaggat 420
ggccttctcc gacggatgtc tcccttgcgt cccgcctccc cttcttgtag gcctgcatca 480
tcaccgtttt tctggacaac cccaaagtac cccgtctccc tggctttagc cacctctcca 540
tcctcttgct ttctttgcct ggacaccccg ttctcctgtg gattcgggtc acctctcact 600
cctttcattt gggcagctcc cctacccccc ttacctctct agtctgtgct agctcttcca 660
gccccctgtc atggcatctt ccaggggtcc gagagctcag ctagtcttct tcctccaacc 720
cgggccccta tgtccacttc aggacagcat gtttgctgcc tccagggatc ctgtgtcccc 780
gagctgggac caccttatat tcccagggcc ggttaactag tacataactt acggtaaatg 840
gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 900
ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 960
ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca 1020
atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta 1080
cttggcagta catctacgta ttagtcatca agcttctcga gggcgcgtga cctcaccctg 1140
tggagccaca ccctagggtt ggccaatcta ctcccaggag cagggagggc aggagccagg 1200
gctgggcata aaagtcaggg cagagccatc tattgcttgc gatcgccacc atgttgaagc 1260
catcattacc attcacatcc ctcttattcc tgcagctgcc cctgctggga gtggggctga 1320
acacgacaat tctgacgccc aatgggaatg aagacaccac agctgatttc ttcctgacca 1380
ctatgcccac tgactccctc agtgtttcca ctctgcccct cccagaggtt cagtgttttg 1440
tgttcaatgt cgagtacatg aattgcactt ggaacagcag ctctgagccc cagcctacca 1500
acctcactct gcattattgg tacaagaact cggataatga taaagtccag aagtgcagcc 1560
actatctatt ctctgaagaa atcacttctg gctgtcagtt gcaaaaaaag gagatccacc 1620
tctaccaaac atttgttgtt cagctccagg acccacggga acccaggaga caggccacac 1680
agatgctaaa actgcagaat ctggtgatcc cctgggctcc agagaaccta acacttcaca 1740
aactgagtga atcccagcta gaactgaact ggaacaacag attcttgaac cactgtttgg 1800
agcacttggt gcagtaccgg actgactggg accacagctg gactgaacaa tcagtggatt 1860
atagacataa gttctccttg cctagtgtgg atgggcagaa acgctacacg tttcgtgttc 1920
ggagccgctt taacccactc tgtggaagtg ctcagcattg gagtgaatgg agccacccaa 1980
tccactgggg gagcaatact tcaaaagaga atcctttcct gtttgcattg gaagccgtgg 2040
ttatctctgt tggctccatg ggattgatta tcagccttct ctgtgtgtat ttctggctgg 2100
aacggacgat gccccgaatt cccaccctga agaacctaga ggatcttgtt actgaatacc 2160
acgggaactt ttcggcctgg agtggtgtgt ctaagggact ggctgagagt ctgcagccag 2220
actacagtga acgactctgc ctcgtcagtg agattccccc aaaaggaggg gcccttgggg 2280
aggggcctgg ggcctcccca tgcaaccagc atagccccta ctgggccccc ccatgttaca 2340
ccctaaagcc tgaaaccgga agcggagcta ctaacttcag cctgctgaag caggctggag 2400
acgtggagga gaaccctgga cctgtcgaca tggtgagcaa gggcgaggag ctgttcaccg 2460
gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt 2520
ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca 2580
ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt 2640
gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg 2700
aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg 2760
ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact 2820
tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg 2880
tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca 2940
acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg 3000
acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag 3060
accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca 3120
ctctcggcat ggacgagctg tacaagtaaa cgcgtgatcc accggatcta gataactgat 3180
cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct 3240
ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc 3300
ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 3360
actgcattct agttgtggtt tgtccaaact catcaatgta tcttaacaga tctatgtggc 3420
tctggttctg ggtactttta tctgtcccct ccaccccaca gtggggccac tagggacagg 3480
attggtgaca gaaaagcccc atccttaggc ctcctccttc ctagtctcct gatattgggt 3540
ctaaccccca cctcctgtta ggcagattcc ttatctggtg acacaccccc atttcctgga 3600
gccatctctc tccttgccag aacctctaag gtttgcttac gatggagcca gagaggatcc 3660
tgggagggag agcttggcag ggggtgggag ggaagggggg gatgcgtgac ctgcccggtt 3720
ctcagtggcc accctgcgct accctctccc agaacctgag ctgctctgac gcggccgtct 3780
ggtgcgtttc actgatcctg gtgctgcagc ttccttacac ttcccaagag gagaagcagt 3840
ttggaaaaac aaaatcagaa taagttggtc ctgagttcta actttggctc ttcacctttc 3900
tagtccccaa tttatattgt tcctccgtgc gtcagtttta cctgtgagat aaggccagta 3960
gccagccccg tcctggcagg gctgtggtga ggaggggggt gtccgtgtgg aaaactccct 4020
ttgtgagaat ggtgcgtcct aggtgttcac caggtcgtgg ccgcctctac tccctttctc 4080
tttctccatc cttctttcct taaagagtcc ccagtgctat ctgggacata ttcctccgcc 4140
cagagcaggg tcccgcttcc ctaaggccct gctctgggct tctgggtttg agtccttggc 4200
aagcccagga gag 4213
<210> 26
<211> 5028
<212> DNA
<213>Artificial sequence (RenGongXuLie)
<400> 26
cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt 60
ggtcgcccgg cctcagtgag cgagcgcgca gagagggagt ggccaactcc atcactaggg 120
gttcctgcgg ccgcacgcgt ctagttatta atagtaatcg aattcgtgtt actcataact 180
agtaaggtcg ggcaggaaga gggcctattt cccatgattc cttcatattt gcatatacga 240
tacaaggctg ttagagagat aattagaatt aatttgactg taaacacaaa gatattagta 300
caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt 360
tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt cttgggttta 420
tatatcttgt ggaaaggacg cgggatccac tggaccaggc agcagcgtca gaagactttt 480
ttggaaaagc ttgactagta atactgtaat agtaatcaat tacggggtca ttagttcata 540
gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 600
ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 660
ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac 720
atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg 780
cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg 840
tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat 900
agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt 960
tttgcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca 1020
aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg 1080
tcagatccgc tagagatccg gtaccgagga gatctgccgc cgcgatcgcc ggcgcgccag 1140
atctcacgct taactagcta gcggaccgac gcgtacgcgg ccgctcgaga tggtgagcaa 1200
gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa 1260
cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac 1320
cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac 1380
cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt 1440
cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga 1500
cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat 1560
cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta 1620
caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt 1680
gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca 1740
gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac 1800
ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt 1860
cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaag tcgaggatta 1920
taaggatgac gacgataaat tcgtcgagca ccaccaccac caccactaat aaggtttatc 1980
cgatccaccg gatctagata agatatccga tccaccggat ctagataact gatcataatc 2040
agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg 2100
aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat 2160
ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat 2220
tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcggtaacca cgtgcggacc 2280
gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg 2340
ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca 2400
gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg 2460
catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg 2520
gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg 2580
ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc 2640
cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc 2700
tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga 2760
cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa 2820
ctggaacaac actcaaccct atctcgggct attcttttga tttataaggg attttgccga 2880
tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca 2940
aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat 3000
agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc 3060
tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt 3120
tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat 3180
aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg 3240
tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga 3300
gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac 3360
atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc 3420
cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca 3480
tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc 3540
caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg 3600
ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac 3660
cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca 3720
taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg 3780
agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac 3840
cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg 3900
caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat 3960
taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg 4020
ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg 4080
cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc 4140
aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc 4200
attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt 4260
tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt 4320
aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt 4380
gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag 4440
cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca 4500
gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca 4560
agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg 4620
ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg 4680
cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct 4740
acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga 4800
gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc 4860
ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg 4920
agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg 4980
cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt 5028

Claims (7)

1. a kind of method using CRISPR/Cas9 system homologous recombination repair IL-2RG dcc genes, it is characterised in that：Pass through PAAV-AAVS1 Donor carriers and pAAV-SaCas9-gRNA AAVS1 carriers are obtained for repairing IL-2RG dcc genes, and After its co-implanted host cell with IL-2RG dcc genes, in pAAV-SaCas9-gRNA AAVS1 carriers CRISPR/Cas9 systems cut defect IL-2RG genes in host cell, and use homologous recombination repair mechanism will Correct IL-2RG genes are properly inserted in genome in pAAV-AAVS1 Donor carriers.
2. a kind of system using CRISPR/Cas9 system homologous recombination repair IL-2RG dcc genes, the system include Two carriers contain CRISPR/Cas9 systems on one of carrier, contain correct IL-2RG genes, institute on another carrier The carrier containing CRISPR/Cas9 systems stated is named as pAAV-SaCas9-gRNA AAVS1 carriers, and nucleotide sequence is such as Shown in SEQ ID No.1, the carrier containing correct IL-2RG genes is named as pAAV-AAVS1Donor carriers, core Nucleotide sequence is as shown in SEQ ID No.2.
3. the pAAV-SaCas9-gRNA AAVS1 carriers described in claim 1-2 are cut for providing CRISPR/Cas9 systems The defects of host cell IL-2RG genes are cut, pAAV-AAVS1 Donor carriers provide correct IL-2RG bases as donor Cause is properly inserted in using homologous recombination repair mechanism in genome.
4. method described in claim 1 is specially：
The defects of host cell is cut using the pAAV-SaCas9-gRNA AAVS1 carriers containing CRISPR/Cas9 systems IL-2RG genes, the pAAV-AAVS1 Donor carriers to contain correct IL-2RG genes provide correct IL-2RG as donor Gene is properly inserted in using homologous recombination repair mechanism in genome.
5. method described in claim 1, more specifically includes the following steps：
(1) pAAV-AAVS1 Donor carriers and pAAV-SaCas9-gRNA AAVS1 carriers are obtained；
(2) pAAV-SaCas9-gRNA AAVS1 carriers and pAAV-AAVS1 Donor carriers are imported into host cell jointly.
6. a kind of kit for repairing IL-2RG dcc genes, the kit includes above-mentioned pAAV-SaCas9-gRNA AAVS1 carriers and pAAV-AAVS1 Donor carriers.
7. the kit described in claim 6 further includes by above-mentioned pAAV-SaCas9-gRNA AAVS1 carriers and pAAV-AAVS1 The required part or all of reagent of Donor vector introduction host cells.
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