质粒名称: | xCas9 3.7 |
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出品公司: | 爱迪基因 |
目录编号: | 108379 |
存储实验室: | David Liu |
载体骨架基本信息 |
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载体名称: | pCMV |
载体出品公司: | -- |
载体抗性: | 氨苄青霉素 |
空载体大小: | -- |
完整质粒大小: | -- |
载体修饰: | 人细胞色素c氧化酶亚基VIII(human cytochrome C oxidase subunit VIII) |
载体类型: | 哺乳细胞表达 |
筛选标记: | 新霉素(G418) |
高拷贝/低拷贝: | 高拷贝 |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | xCas9 3.7 |
物种来源: | -- |
插入大小: | 4140 bp |
基因突变: | -- |
GenBank编号 | -- |
启动子 | -- |
标签/融合蛋白: | 人细胞色素c氧化酶亚基VIII(N端) |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | AgeI(未破坏) |
3' 克隆位点: | NotI (未破坏) |
5' 测序引物 | CMV-F |
3' 测序引物 | SV40pA-R |
文献引用方法 |
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材料和方法部分: | xCas9 3.7 was a gift from David Liu (Addgene plasmid # 108379 ; http://n2t.net/addgene:108379 ; RRID:Addgene_108379) |
参考文献部分: | Evolved Cas9 variants with broad PAM compatibility and high DNA specificity. Hu JH, Miller SM, Geurts MH, Tang W, Chen L, Sun N, Zeina CM, Gao X, Rees HA, Lin Z, Liu DR. Nature. 2018 Feb 28. pii: nature26155. doi: 10.1038/nature26155. 10.1038/nature26155 PubMed 29512652 |
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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 xCas9 3.7 9553 bp ds-DNA circular SYN 16-APR-2019 DEFINITION Mammalian expression vector for xCas9 3.7. ACCESSION . VERSION . KEYWORDS xCas9 3.7 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9553) AUTHORS Hu JH, Miller SM, Geurts MH, Tang W, Chen L, Sun N, Zeina CM, Gao X, Rees HA, Lin Z, Liu DR TITLE Evolved Cas9 variants with broad PAM compatibility and high DNA specificity. JOURNAL Nature. 2018 Feb 28. pii: nature26155. doi: 10.1038/nature26155. PUBMED 29512652 REFERENCE 2 (bases 1 to 9553) AUTHORS . TITLE Direct Submission JOURNAL Exported Apr 16, 2019 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9553 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 50..429 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 430..633 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 584..604 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 630..654 /label=LNCX /note="Human CMV promoter, forward primer" regulatory 742..751 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 748..4851 /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 ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDTKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKLYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL RKQRTFDNGIIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG NSRFAWMTRKSEETITPWNFEKVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY FTVYNELTKVKYVTEGMRKPAFLSGDQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFI QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGVLQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGD" CDS 4864..4884 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" primer_bind complement(4939..4958) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4983..5031 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5233..5661 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5320..5339) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5530..5551 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5670..5690) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 5675..6004 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 5855..5990 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5917..5936 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 6071..6865 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(6125..6144) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6735..6754 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 7041..7162 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(7078..7097) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 7132..7151 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(7211..7227) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7211..7227) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(7224..7246) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 7235..7251 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(7259..7289) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7304..7325 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7442..7459) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7613..8201) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7693..7712) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8372..9232) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 8995..9014 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(9233..9337) /gene="bla" /label=AmpR promoter primer_bind complement(9412..9431) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta 61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 601 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg 661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg 721 actctagagg atcgaaccct tgccaccatg gacaagaagt actccattgg gctcgatatc 781 ggcacaaaca gcgtcggctg ggccgtcatt acggacgagt acaaggtgcc gagcaaaaaa 841 ttcaaagttc tgggcaatac cgatcgccac agcataaaga agaacctcat tggcgccctc 901 ctgttcgact ccggggagac ggccgaagcc acgcggctca aaagaacagc acggcgcaga 961 tatacccgca gaaagaatcg gatctgctac ctgcaggaga tctttagtaa tgagatggct 1021 aaggtggatg actctttctt ccataggctg gaggagtcct ttttggtgga ggaggataaa 1081 aagcacgagc gccacccaat ctttggcaat atcgtggacg aggtggcgta ccatgaaaag 1141 tacccaacca tatatcatct gaggaagaag cttgtagaca gtactgataa ggctgacttg 1201 cggttgatct atctcgcgct ggcgcatatg atcaaatttc ggggacactt cctcatcgag 1261 ggggacctga acccagacaa cagcgatgtc gacaaactct ttatccaact ggttcagact 1321 tacaatcagc ttttcgaaga gaacccgatc aacgcatccg gagttgacgc caaagcaatc 1381 ctgagcgcta ggctgtccaa atcccggcgg ctcgaaaacc tcatcgcaca gctccctggg 1441 gagaagaaga acggcctgtt tggtaatctt atcgccctgt ccctcgggct gacccccaac 1501 tttaaatcta acttcgacct ggccgaagat accaagcttc aactgagcaa agacacctac 1561 gatgatgatc tcgacaatct gctggcccag atcggcgacc agtacgcaga cctttttttg 1621 gcggcaaaga acctgtcaga cgccattctg ctgagtgata ttctgcgagt gaacacggag 1681 atcaccaaag ctccgctgag cgctagtatg atcaagctct atgatgagca ccaccaagac 1741 ttgactttgc tgaaggccct tgtcagacag caactgcctg agaagtacaa ggaaattttc 1801 ttcgatcagt ctaaaaatgg ctacgccgga tacattgacg gcggagcaag ccaggaggaa 1861 ttttacaaat ttattaagcc catcttggaa aaaatggacg gcaccgagga gctgctggta 1921 aagcttaaca gagaagatct gttgcgcaaa cagcgcactt tcgacaatgg aatcatcccc 1981 caccagattc acctgggcga actgcacgct atcctcaggc ggcaagagga tttctacccc 2041 tttttgaaag ataacaggga aaagattgag aaaatcctca catttcggat accctactat 2101 gtaggccccc tcgcccgggg aaattccaga ttcgcgtgga tgactcgcaa atcagaagag 2161 accatcactc cctggaactt cgagaaagtc gtggataagg gggcctctgc ccagtccttc 2221 atcgaaagga tgactaactt tgataaaaat ctgcctaacg aaaaggtgct tcctaaacac 2281 tctctgctgt acgagtactt cacagtttat aacgagctca ccaaggtcaa atacgtcaca 2341 gaagggatga gaaagccagc attcctgtct ggagatcaga agaaagctat tgtggacctc 2401 ctcttcaaga cgaaccggaa agttaccgtg aaacagctca aagaagacta tttcaaaaag 2461 attgaatgtt tcgactctgt tgaaatcagc ggagtggagg atcgcttcaa cgcatccctg 2521 ggaacgtatc acgatctcct gaaaatcatt aaagacaagg acttcctgga caatgaggag 2581 aacgaggaca ttcttgagga cattgtcctc acccttacgt tgtttgaaga tagggagatg 2641 attgaagaac gcttgaaaac ttacgctcat ctcttcgacg acaaagtcat gaagcagctc 2701 aagaggcgcc gatatacagg atgggggcgg ctgtcaagaa aactgatcaa tgggatccga 2761 gacaagcaga gtggaaagac aatcctggat tttcttaagt ccgatggatt tgccaaccgg 2821 aacttcattc agttgatcca tgatgactct ctcaccttta aggaggacat ccagaaagca 2881 caagtttctg gccaggggga cagtcttcac gagcacatcg ctaatcttgc aggtagccca 2941 gctatcaaaa agggaatact gcagaccgtt aaggtcgtgg atgaactcgt caaagtaatg 3001 ggaaggcata agcccgagaa tatcgttatc gagatggccc gagagaacca aaccacccag 3061 aagggacaga agaacagtag ggaaaggatg aagaggattg aagagggtat aaaagaactg 3121 gggtcccaaa tccttaagga acacccagtt gaaaacaccc agcttcagaa tgagaagctc 3181 tacctgtact acctgcagaa cggcagggac atgtacgtgg atcaggaact ggacatcaat 3241 cggctctccg actacgacgt ggaccatatc gtgccccagt cttttctcaa agatgattct 3301 attgataata aagtgttgac aagatccgat aaaaacagag ggaagagtga taacgtcccc 3361 tcagaagaag ttgtcaagaa aatgaaaaat tattggcggc agctgctgaa cgccaaactg 3421 atcacacaac ggaagttcga taatctgact aaggctgaac gaggtggcct gtctgagttg 3481 gataaagccg gtttcatcaa aaggcagctt gttgagacac gccagatcac caagcacgtg 3541 gcccaaattc tcgattcacg catgaacacc aagtacgatg aaaatgacaa actgattcga 3601 gaggtgaaag ttattactct gaagtctaag ctggtctcag atttcagaaa ggactttcag 3661 ttttataagg tgagagagat caacaattac caccatgcgc atgatgccta cctgaatgca 3721 gtggtaggca ctgcacttat caaaaaatat cccaagcttg aatctgaatt tgtttacgga 3781 gactataaag tgtacgatgt taggaaaatg atcgcaaagt ctgagcagga aataggcaag 3841 gccaccgcta agtacttctt ttacagcaat attatgaatt ttttcaagac cgagattaca 3901 ctggccaatg gagagattcg gaagcgacca cttatcgaaa caaacggaga aacaggagaa 3961 atcgtgtggg acaagggtag ggatttcgcg acagtccgga aggtcctgtc catgccgcag 4021 gtgaacatcg ttaaaaagac cgaagtacag accggaggtt tctccaagga aagtatcctc 4081 ccgaaaagga acagcgacaa gctgatcgca cgcaaaaaag attgggaccc caagaaatac 4141 ggcggattcg attctcctac agtcgcttac agtgtactgg ttgtggccaa agtggagaaa 4201 gggaagtcta aaaaactcaa aagcgtcaag gaactgctgg gcatcacaat catggagcga 4261 tcaagcttcg aaaaaaaccc catcgacttt ctcgaggcga aaggatataa agaggtcaaa 4321 aaagacctca tcattaagct tcccaagtac tctctctttg agcttgaaaa cggccggaaa 4381 cgaatgctcg ctagtgcggg cgtgctgcag aaaggtaacg agctggcact gccctctaaa 4441 tacgttaatt tcttgtatct ggccagccac tatgaaaagc tcaaagggtc tcccgaagat 4501 aatgagcaga agcagctgtt cgtggaacaa cacaaacact accttgatga gatcatcgag 4561 caaataagcg aattctccaa aagagtgatc ctcgccgacg ctaacctcga taaggtgctt 4621 tctgcttaca ataagcacag ggataagccc atcagggagc aggcagaaaa cattatccac 4681 ttgtttactc tgaccaactt gggcgcgcct gcagccttca agtacttcga cactaccata 4741 gacagaaagc ggtacacctc tacaaaggag gtcctggacg ccacactgat tcatcagtca 4801 attacggggc tctatgaaac aagaatcgac ctctctcagc tcggtggaga cagcagggct 4861 gaccccaaga agaagaggaa ggtgtgaaag ggttcgatcc ctaccggtta gtaatgagtt 4921 taaacggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat 4981 gacggcaata aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg 5041 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat 5101 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc 5161 tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc tggggctcta 5221 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc 5281 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt 5341 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag 5401 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt 5461 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt 5521 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt 5581 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt 5641 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc 5701 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag 5761 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta 5821 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc 5881 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc 5941 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg 6001 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg 6061 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga 6121 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt 6181 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct 6241 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg 6301 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt 6361 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc 6421 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc 6481 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga 6541 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg 6601 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat 6661 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg 6721 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc 6781 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta 6841 tcgccttctt gacgagttct tctgagcggg actctggggt tcgcgaaatg accgaccaag 6901 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg 6961 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc 7021 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca 7081 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt 7141 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg 7201 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca 7261 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca 7321 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc 7381 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt 7441 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact 7501 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag 7561 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata 7621 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc 7681 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg 7741 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc 7801 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg 7861 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc 7921 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga 7981 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg 8041 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa 8101 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg 8161 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt 8221 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat 8281 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct 8341 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta 8401 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa 8461 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac 8521 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa 8581 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag 8641 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg 8701 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag 8761 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg 8821 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc 8881 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat 8941 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata 9001 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa 9061 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca 9121 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc 9181 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc 9241 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg 9301 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac 9361 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg 9421 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga 9481 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa 9541 gaatctgctt agg //
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