质粒名称: | pWS172 |
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出品公司: | Addgene |
目录编号: | 90519 |
存储实验室: | Tom Ellis |
载体骨架基本信息 |
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载体名称: | 2 micron |
空载体大小: | 3882 bp |
完整质粒大小: | 11132bp |
载体类型: | Yeast Expression, CRISPR |
筛选标记: | HIS3 |
载体抗性: | Kanamycin |
生长温度: | 37 ℃ |
宿主菌: | NEB Stable |
拷贝数: | 高拷贝 |
插入基因信息 |
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插入基因名称: | Cas9 |
物种来源: | S. cerevisiae (budding yeast) |
文献引用方法 |
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材料和方法部分: | pWS172 was a gift from Tom Ellis (Addgene plasmid # 90519 ; http://n2t.net/addgene:90519 ; RRID:Addgene_90519) |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pWS172 11132 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Cas9 gap repair vector - HIS3. ACCESSION . VERSION . KEYWORDS pWS172 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11132) TITLE Ellis Lab Yeast CRISPR Plasmids JOURNAL Unpublished REFERENCE 2 (bases 1 to 11132) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..11132 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 1..589 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 490..509 /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 1451..5554 /codon_start=1 /product="Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=Cas9 /note="generates RNA-guided double strand breaks in DNA" /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGD" CDS 5567..5587 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 5591..5611 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" RBS 6521..6532 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" CDS 6539..7252 /codon_start=1 /product="GFP variant that folds robustly even when fused to poorly folded proteins (Pedelacq et al., 2006)" /label=superfolder GFP /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE FVTAAGITHGMDELYK" terminator 7264..7335 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 7351..7378 /label=T7Te terminator /note="phage T7 early transcription terminator" misc_RNA 7392..7467 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" promoter 7988..8175 /gene="S. cerevisiae HIS3" /label=HIS3 promoter CDS 8176..8838 /codon_start=1 /gene="S. cerevisiae HIS3" /product="imidazoleglycerol-phosphate dehydratase, required for histidine biosynthesis" /label=HIS3 /note="yeast auxotrophic marker" /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM" rep_origin 9053..10035 /label=2u ori /note="yeast 2u plasmid origin of replication" CDS complement(10093..10908) /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind 10816..10835 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 10900..10919 /label=pENTR-R /note="pENTR vectors, reverse primer" ORIGIN 1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc 61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt 121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt 181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg 421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc 661 gttatcccct gattctgtgg ataaccgtgc ggccgcccct gaattcgcat ctagactgaa 721 ctggccgata attgcagacg aacggtgagt aaggaaagag tgaggaacta tcgcatacct 781 gcatttaaag atgccgattt gggcgcgaat cctttatttt ggcttcaccc tcatactatt 841 atcagggcca gaaaaaggaa gtgtttccct ccttcttgaa ttgatgttac cctcataaag 901 cacgtggcct cttatcgaga aagaaattac cgtcgctcgt gatttgtttg caaaaagaac 961 aaaactgaaa aaacccagac acgctcgact tcctgtcatc ctattgattg cagcttccaa 1021 tttcgtcaca caacaaggtc ctagcgacgg ctcacaggtt ttgtaacaag caatcgaagg 1081 ttctggaatg gcgggaaagg gtttagtacc acatgctatg atgcccactg tgatctccag 1141 agcaaagttc gttcgatcgt actgttactc tctctctttc aaacagaatt gtccgaatcg 1201 tgtgacaaca acagcctgtt ctcacacact cttttcttct aaccaagggg gtggtttagt 1261 ttagtagaac ctcgtgaaac ttacatttac atatatataa acttgcataa attggtcaat 1321 gcaagaaata catatttggt cttttctaat tcgtagtttt tcaagttctt agatgctttc 1381 tttttctctt ttttacagat catcaaggaa gtaattatct actttttaca acaaatataa 1441 aacaagatct atggacaaga agtattctat cggactggat atcgggacta atagcgtcgg 1501 gtgggccgtg atcactgacg agtacaaggt gccctctaag aagttcaagg tgctcgggaa 1561 caccgaccgg cattccatca agaaaaatct gatcggagct ctcctctttg attcagggga 1621 aaccgctgaa gcaacccgcc tcaagcggac tgctagacgg cggtacacca ggaggaagaa 1681 ccggatttgt taccttcaag agatattctc caacgaaatg gcaaaggtcg acgacagctt 1741 cttccatagg ctggaagaat cattcctcgt ggaagaggat aagaagcatg aacggcatcc 1801 catcttcggt aatatcgtcg acgaggtggc ctatcacgag aaatacccaa ccatctacca 1861 tcttcgcaaa aagctggtgg actcaaccga caaggcagac ctccggctta tctacctggc 1921 cctggcccac atgatcaagt tcagaggcca cttcctgatc gagggcgacc tcaatcctga 1981 caatagcgat gtggataaac tgttcatcca gctggtgcag acttacaacc agctctttga 2041 agagaacccc atcaatgcaa gcggagtcga tgccaaggcc attctgtcag cccggctgtc 2101 aaagagccgc agacttgaga atcttatcgc tcagctgccg ggtgaaaaga aaaatggact 2161 gttcgggaac ctgattgctc tttcacttgg gctgactccc aatttcaagt ctaatttcga 2221 cctggcagag gatgccaagc tgcaactgtc caaggacacc tatgatgacg atctcgacaa 2281 cctcctggcc cagatcggtg accaatacgc cgaccttttc cttgctgcta agaatctttc 2341 tgacgccatc ctgctgtctg acattctccg cgtgaacact gaaatcacca aggcccctct 2401 ttcagcttca atgattaagc ggtatgatga gcaccaccag gacctgaccc tgcttaaggc 2461 actcgtccgg cagcagcttc cggagaagta caaggaaatc ttctttgacc agtcaaagaa 2521 tggatacgcc ggctacatcg acggaggtgc ctcccaagag gaattttata agtttatcaa 2581 acctatcctt gagaagatgg acggcaccga agagctcctc gtgaaactga atcgggagga 2641 tctgctgcgg aagcagcgca ctttcgacaa tgggagcatt ccccaccaga tccatcttgg 2701 ggagcttcac gccatccttc ggcgccaaga ggacttctac ccctttctta aggacaacag 2761 ggagaagatt gagaaaattc tcactttccg catcccctac tacgtgggac ccctcgccag 2821 aggaaatagc cggtttgctt ggatgaccag aaagtcagaa gaaactatca ctccctggaa 2881 cttcgaagag gtggtggaca agggagccag cgctcagtca ttcatcgaac ggatgactaa 2941 cttcgataag aacctcccca atgagaaggt cctgccgaaa cattccctgc tctacgagta 3001 ctttaccgtg tacaacgagc tgaccaaggt gaaatatgtc accgaaggga tgaggaagcc 3061 cgcattcctg tcaggcgaac aaaagaaggc aattgtggac cttctgttca agaccaatag 3121 aaaggtgacc gtgaagcagc tgaaggagga ctatttcaag aaaattgaat gcttcgactc 3181 tgtggagatt agcggggtcg aagatcggtt caacgcaagc ctgggtactt accatgatct 3241 gcttaagatc atcaaggaca aggattttct ggacaatgag gagaacgagg acatccttga 3301 ggacattgtc ctgactctca ctctgttcga ggaccgggaa atgatcgagg agaggcttaa 3361 gacctacgcc catctgttcg acgataaagt gatgaagcaa cttaaacgga gaagatatac 3421 cggatgggga cgccttagcc gcaaactcat caacggaatc cgggacaaac agagcggaaa 3481 gaccattctt gatttcctta agagcgacgg attcgctaat cgcaacttca tgcaacttat 3541 ccatgatgat tccctgacct ttaaggagga catccagaag gcccaagtgt ctggacaagg 3601 tgactcactg cacgagcata tcgcaaatct ggctggttca cccgctatta agaagggtat 3661 tctccagacc gtgaaagtcg tggacgagct ggtcaaggtg atgggtcgcc ataaaccaga 3721 gaacattgtc atcgagatgg ccagggaaaa ccagactacc cagaagggac agaagaacag 3781 cagggagcgg atgaaaagaa ttgaggaagg gattaaggag ctcgggtcac agatccttaa 3841 agagcacccg gtggaaaaca cccagcttca gaatgagaag ctctatctgt actaccttca 3901 aaatggacgc gatatgtatg tggaccaaga gcttgatatc aacaggctct cagactacga 3961 cgtggaccat atcgtccctc agagcttcct caaagacgac tcaattgaca ataaggtgct 4021 gactcgctca gacaagaacc ggggaaagtc agataacgtg ccctcagagg aagtcgtgaa 4081 aaagatgaag aactattggc gccagcttct gaacgcaaag ctgatcactc agcggaagtt 4141 cgacaatctc actaaggctg agaggggcgg actgagcgaa ctggacaaag caggattcat 4201 taaacggcaa cttgtggaga ctcggcagat tactaaacat gtcgcccaaa tccttgactc 4261 acgcatgaat accaagtacg acgaaaacga caaacttatc cgcgaggtga aggtgattac 4321 cctgaagtcc aagctggtca gcgatttcag aaaggacttt caattctaca aagtgcggga 4381 gatcaataac tatcatcatg ctcatgacgc atatctgaat gccgtggtgg gaaccgccct 4441 gatcaagaag tacccaaagc tggaaagcga gttcgtgtac ggagactaca aggtctacga 4501 cgtgcgcaag atgattgcca aatctgagca ggagatcgga aaggccaccg caaagtactt 4561 cttctacagc aacatcatga atttcttcaa gaccgaaatc acccttgcaa acggtgagat 4621 ccggaagagg ccgctcatcg agactaatgg ggagactggc gaaatcgtgt gggacaaggg 4681 cagagatttc gctaccgtgc gcaaagtgct ttctatgcct caagtgaaca tcgtgaagaa 4741 aaccgaggtg caaaccggag gcttttctaa ggaatcaatc ctccccaagc gcaactccga 4801 caagctcatt gcaaggaaga aggattggga ccctaagaag tacggcggat tcgattcacc 4861 aactgtggct tattctgtcc tggtcgtggc taaggtggaa aaaggaaagt ctaagaagct 4921 caagagcgtg aaggaactgc tgggtatcac cattatggag cgcagctcct tcgagaagaa 4981 cccaattgac tttctcgaag ccaaaggtta caaggaagtc aagaaggacc ttatcatcaa 5041 gctcccaaag tatagcctgt tcgaactgga gaatgggcgg aagcggatgc tcgcctccgc 5101 tggcgaactt cagaagggta atgagctggc tctcccctcc aagtacgtga atttcctcta 5161 ccttgcaagc cattacgaga agctgaaggg gagccccgag gacaacgagc aaaagcaact 5221 gtttgtggag cagcataagc attatctgga cgagatcatt gagcagattt ccgagttttc 5281 taaacgcgtc attctcgctg atgccaacct cgataaagtc cttagcgcat acaataagca 5341 cagagacaaa ccaattcggg agcaggctga gaatatcatc cacctgttca ccctcaccaa 5401 tcttggtgcc cctgccgcat tcaagtactt cgacaccacc atcgaccgga aacgctatac 5461 ctccaccaaa gaagtgctgg acgccaccct catccaccag agcatcaccg gactttacga 5521 aactcggatt gacctctcac agctcggagg ggatgaggga gctgatccaa aaaagaagag 5581 aaaggtagat ccaaaaaaga agagaaaggt ataaggatcc taactcgaga ttgaattgaa 5641 ttgaaatcga tagatcaatt tttttctttt ctctttcccc atcctttacg ctaaaataat 5701 agtttatttt attttttgaa tattttttat ttatatacgt atatatagac tattatttat 5761 cttttaatga ttattaagat ttttattaaa aaaaaattcg ctcctctttt aatgccttta 5821 tgcagttttt ttttcccatt cgatatttct atgtgctgcc aagatatcaa caacagagga 5881 catatgccct acctccatga tctactgacg tccctgaggc tgcaattcat gtaatggggc 5941 agtatccgcg gcaagtccta gtgcaatggc ggttttttac cctcgttctg aagaagaggc 6001 gacgcgggtg cggtcatcac taatgtggaa attggtaaga ctctcgggcc tccgccttta 6061 ggcggtgctt actctttcat aaaggggctg ttagttatgc aacgatgtgc ttcagtatta 6121 cattttttgc cttcaacgcc ttgattgttc tatttttgct aataataaat ctatttcatc 6181 ggactaaaag tccattagtt gtaagcggat ttagctcagt tgggagagcg ccagactgaa 6241 gaaaaacttc ggtcaagtca tctggaggtc ctgtgttcga tccacagaat tcgcagatgg 6301 ccggcatggt cccagcctcc tcgctggcgc cggctgggca acaccttcgg gtggcgaatg 6361 ggacttgaga cggaaagtga aacgtgattt catgcgtcat tttgaacatt ttgtaaatct 6421 tatttaataa tgtgtgcggc aattcacatt taatttatga atgttttctt aacatcgcgg 6481 caactcaaga aacggcaggt tcggatctta gctactagag aaagaggaga aatactagat 6541 gcgtaaaggc gaagagctgt tcactggtgt cgtccctatt ctggtggaac tggatggtga 6601 tgtcaacggt cataagtttt ccgtgcgtgg cgagggtgaa ggtgacgcaa ctaatggtaa 6661 actgacgctg aagttcatct gtactactgg taaactgccg gttccttggc cgactctggt 6721 aacgacgctg acttatggtg ttcagtgctt tgctcgttat ccggaccata tgaagcagca 6781 tgacttcttc aagtccgcca tgccggaagg ctatgtgcag gaacgcacga tttcctttaa 6841 ggatgacggc acgtacaaaa cgcgtgcgga agtgaaattt gaaggcgata ccctggtaaa 6901 ccgcattgag ctgaaaggca ttgactttaa agaggacggc aatatcctgg gccataagct 6961 ggaatacaat tttaacagcc acaatgttta catcaccgcc gataaacaaa aaaatggcat 7021 taaagcgaat tttaaaattc gccacaacgt ggaggatggc agcgtgcagc tggctgatca 7081 ctaccagcaa aacactccaa tcggtgatgg tcctgttctg ctgccagaca atcactatct 7141 gagcacgcaa agcgttctgt ctaaagatcc gaacgagaaa cgcgatcata tggttctgct 7201 ggagttcgta accgcagcgg gcatcacgca tggtatggat gaactgtaca aatgaccagg 7261 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg 7321 tcggtgaacg ctctctacta gagtcacact ggctcacctt cgggtgggcc tttctgcgtt 7381 tatacgtctc agttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa 7441 cttgaaaaag tggcaccgag tcggtgcttt ttttattttt tgtcactatt gttatgtaaa 7501 atgccacctc tgacagtatg gaacgcaaac ttctgtctag tggatagctg gacattccag 7561 gcagtgcctc tgccgccgga cccctctcgt gattgggtag ctggacatgc ccttgtaaga 7621 tataacaaga gcctgcctgt ctaatgatct cacggcgaaa gtcggggaga cagcagcggc 7681 tgcagacatt ataccgcaac aacactaagg tgagataact ccgtacattg actacgcgtt 7741 cctctagacc ttacttgacc ggatacagtg tctttgacac gtttatgggt tacagcaatc 7801 acatccaaga ctggctatgc acgaagcaac tcttgagtgt taaaatgttg acccctgtat 7861 ttgggatgcg ggtagtagat gagtgcagga tatcagcagt tacagagatg ttacgaacca 7921 ctagtgcact gcagtacaaa cacagtcctt tcccgcaatt ttctttttct attactcttg 7981 gcctcctcta gtacactcta tattttttta tgcctcggta atgattttca tttttttttt 8041 tccacctagc ggatgactct ttttttttct tagcgattgg cattatcaca taatgaatta 8101 tacattatat aaagtaatgt gatttcttcg aagaatatac taaaaaatga gcaggcaaga 8161 taaacgaagg caaagatgac agagcagaaa gccctagtaa agcgtattac aaatgaaacc 8221 aagattcaga ttgcgatctc tttaaagggt ggtcccctag cgatagagca ctcgatcttc 8281 ccagaaaaag aggcagaagc agtagcagaa caggccacac aatcgcaagt gattaacgtc 8341 cacacaggta tagggtttct ggaccatatg atacatgctc tggccaagca ttccggctgg 8401 tcgctaatcg ttgagtgcat tggtgactta cacatagacg accatcacac cactgaggac 8461 tgcgggattg ctctcggtca agcttttaaa gaggccctag gggccgtgcg tggagtaaaa 8521 aggtttggat caggatttgc gcctttggat gaggcacttt ccagagcggt ggttgatctt 8581 tcgaacaggc cgtacgcagt tgtcgaactt ggtttgcaaa gggagaaagt aggtgatctc 8641 tcttgcgaga tgatcccgca ttttcttgaa agctttgcag aggctagcag aattaccctc 8701 cacgttgatt gtctgcgagg caagaatgat catcaccgta gtgagagtgc gttcaaggct 8761 cttgcggttg ccataagaga agccacctcg cccaatggta ccaacgatgt tccctccacc 8821 aaaggtgttc ttatgtagtg acaccgatta tttaaagctg ctgcatacga tatatataca 8881 tgtgtatata tgtataccta tgaatgtcag taagtatgta tacgaacagt atgatactga 8941 agatgacaag gtaatgcatc attctatacg tgtcattctg aacgaggcgc gctttccttt 9001 tttctttttg ctttttcttt ttttttctct tgaactcgac ggatcataga gtaacgaagc 9061 atctgtgctt cattttgtag aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa 9121 gaatctgagc tgcattttta cagaacagaa atgcaacgcg aaagcgctat tttaccaacg 9181 aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa 9241 acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgagagc gctattttac 9301 caacaaagaa tctatacttc ttttttgttc tacaaaaatg catcccgaga gcgctatttt 9361 tctaacaaag catcttagat tacttttttt ctcctttgtg cgctctataa tgcagtctct 9421 tgataacttt ttgcactgta ggtccgttaa ggttagaaga aggctacttt ggtgtctatt 9481 ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt tactgattac tagcgaagct 9541 gcgggtgcat tttttcaaga taaaggcatc cccgattata ttctataccg atgtggattg 9601 cgcatacttt gtgaacagaa agtgatagcg ttgatgattc ttcattggtc agaaaattat 9661 gaacggtttc ttctattttg tctctatata ctacgtatag gaaatgttta cattttcgta 9721 ttgttttcga ttcactctat gaatagttct tactacaatt tttttgtcta aagagtaata 9781 ctagagataa acataaaaaa tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag 9841 gtggatgggt aggttatata gggatatagc acagagatat atagcaaaga gatacttttg 9901 agcaatgttt gtggaagcgg tattcgcaat attttagtag ctcgttacag tccggtgcgt 9961 ttttggtttt ttgaaagtgc gtcatcagag cgcttttggt tttcaaaagc gctctgaagt 10021 tcctatactt tctagctaga gaataggaac ttcccgagcg gccgcgtgtt acaaccaatt 10081 aaccaattct gattagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc 10141 aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc 10201 gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac 10261 atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc 10321 atgagtgacg actgaatccg gtgagaatgg caaaagctta tgcatttctt tccagacttg 10381 ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt 10441 cattcgtgat tgcgcctgag cgaggcgaaa tacgcgatcg ctgttaaaag gacaattaca 10501 aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc 10561 tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag 10621 taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc 10681 cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc 10741 atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc 10801 tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga 10861 atttaatcgc ggcctggagc aagacgtttc ccgttgaata tggctcataa caccccttgt 10921 attactgttt atgtaagcag acagttttat tgttcatgat gatatatttt tatcttgtgc 10981 aatgtaacat cagagatttt gagacacaac gtggctttgt tgaataaatc gaacttttgc 11041 tgagttgaag gatcagtcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg 11101 tcagaccccg tagaaaagat caaaggatct tc //
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