质粒名称: | pEPSA5Cas9syn2 |
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出品公司: | Addgene |
目录编号: | 206963 |
存储实验室: | Daijie Chen |
体骨架基本信息 | |
载体名称: | pEPSA5 |
载体出品公司: | Clontech |
空载体大小: | 5500bp |
完整质粒大小: | 10062bp |
载体类型: | Bacterial Expression, CRISPR, Synthetic Biology |
载体抗性: | 氨苄霉素 |
生长温度: | 37 ℃ |
宿主菌: | DH5a |
拷贝数: | 低拷贝 |
插入基因信息 |
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克隆方法: | Ligation Independent Cloning |
文献引用方法 |
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材料和方法部分: | pEPSA5Cas9syn2 was a gift from Daijie Chen (Addgene plasmid # 206963 ; http://n2t.net/addgene:206963 ; RRID:Addgene_206963) |
参考文献部分: | Efficient Genome Editing in Most Staphylococcus aureus by Using the Restriction-Modification System Silent CRISPR-Cas9 Toolkit. Yang P, Yang J, Lin T, Liu Q, Yin Y, Chen D, Yang S. ACS Synth Biol. 2023 Oct 13. doi: 10.1021/acssynbio.3c00339. 10.1021/acssynbio.3c00339 PubMed 37830328 |
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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 pEPSA5Cas9syn2 10062 bp ds-DNA circular SYN 10-NOV-2023 DEFINITION S. aureus genome editing vector removed these two motifs AGGNNNNNGAT, CCAYNNNNNNTGT, CCAYNNNNNNGTA, GGANNNNNNNNTCG, and GWAGNNNNNNGAT. ACCESSION . VERSION . KEYWORDS pEPSA5Cas9syn2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10062) TITLE Efficient Genome Editing in Most Staphylococcus aureus by Using the Restriction-modification Systems Silent CRISPR-Cas9 Toolkit JOURNAL Unpublished REFERENCE 2 (bases 1 to 10062) AUTHORS . TITLE Direct Submission JOURNAL Exported Nov 10, 2023 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..10062 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin complement(260..805) /direction=LEFT /label=p15A ori /note="Plasmids containing the medium-copy-number p15A origin of replication can be propagated in E. coli cells that contain a second plasmid with the ColE1 origin. " primer_bind complement(340..359) /label=pBR322ori-F /note="pBR322 origin, forward primer" misc_RNA 4102..4177 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" CDS 4418..8524 /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" primer_bind complement(8555..8572) /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" terminator 8725..8811 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 8903..8930 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" CDS complement(9072..9932) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 9695..9714 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(9933..10037) /gene="bla" /label=AmpR promoter ORIGIN 1 ggcggccgca ctggcttact atgttggcac tgatgagggt gtcagtgaag tgcttcatgt 61 ggcaggagaa aaaaggctgc accggtgcgt cagcagaata tgtgatacag gatatattcc 121 gcttcctcgc tcactgactc gctacgctcg gtcgttcgac tgcggcgagc ggaaatggct 181 tacgaacggg gcggagattt cctggaagat gccaggaaga tacttaacag ggaagtgaga 241 gggccgcggc aaagccgttt ttccataggc tccgcccccc tgacaagcat cacgaaatct 301 gacgctcaaa tcagtggtgg cgaaacccga caggactata aagataccag gcgtttcccc 361 ctggcggctc cctcgtgcgc tctcctgttc ctgcctttcg gtttaccggt gtcattccgc 421 tgttatggcc gcgtttgtct cattccacgc ctgacactca gttccgggta ggcagttcgc 481 tccaagctgg actgtatgca cgaacccccc gttcagtccg accgctgcgc cttatccggt 541 aactatcgtc ttgagtccaa cccggaaaga catgcaaaag caccactggc agcagccact 601 ggtaattgat ttagaggagt tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag 661 gacaagtttt ggtgactgcg ctcctccaag ccagttacct cggttcaaag agttggtagc 721 tcagagaacc ttcgaaaaac cgccctgcaa ggcggttttt tcgttttcag agcaagagat 781 tacgcgcaga ccaaaacgat ctcaagaaga tcatcttatg cggccgcttc tttcctgcgt 841 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc 901 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc 961 ggtattttct ccttacgcat ctgtgcggta tttcacaccg cataggtaga tccctcgacc 1021 tgcaggcatg caagcttctg taggttttta ggcataaaac tatatgattt acccctaaat 1081 ctttaaaatg ccccttaaaa ttcaaaataa aggcatttaa aatttaaata tttcttgtga 1141 taaagtttgt taaaaaggag tggttttatg actgttatgt ggttatcgat tataggtatg 1201 tggttttgta ttggaatggc attttttgct atcaaggtta ttaaaaataa aaattagacc 1261 acgcatttat gccgagaaaa tttattgtgc gttgagaaga acccttaact aaacttgcag 1321 acgaatgtcg gcatagcgtg agctattaag ccgaccattc gacaagtttt gggattgtta 1381 agggttccga ggctcaacgt caataaagca attggaataa agaagcgaaa aaggagaagt 1441 cggttcagaa aaagaagcat atgcatctgg agctgtaata taaaaacctt cttcaactaa 1501 cggggcaggt tagtgacatt agaaaaccga ctgtaaaaag tacagtcggc attatctcat 1561 attataaaag ccagtcatta ggcctatctg acaattcctg aatagagttc ataaacaatc 1621 ctgcatgata accatcacaa acagaatgat gtacctgtaa agatagcggt aaatatattg 1681 aattaccttt attaatgaat tttcctgctg taataatggg tagaaggtaa ttactattat 1741 tattgatatt taagttaaac ccagtaaatg aagtccatgg aataatagaa agagaaaaag 1801 cattttcagg tataggtgtt ttgggaaaca atttccccga accattatat ttctctacat 1861 cagaaaggta taaatcataa aactctttga agtcattctt tacaggagtc caaataccag 1921 agaatgtttt agatacacca tcaaaaattg tataaagtgg ctctaactta tcccaataac 1981 ctaactctcc gtcgctattg taaccagttc taaaagctgt atttgagttt atcacccttg 2041 tcactaagaa aataaatgca gggtaaaatt tatatccttc ttgttttatg tttcggtata 2101 aaacactaat atcaatttct gtggttatac taaaagtcgt ttgttggttc aaataatgat 2161 taaatatctc ttttctcttc caattgtcta aatcaatttt attaaagttc atttgatatg 2221 cctcctaaat ttttatctaa agtgaattta ggaggcttac ttgtctgctt tcttcattag 2281 aatcaatcct tttttaaaag tcaatattac tgtaacataa atatatattt taaaaatatc 2341 ccactttatc caattttcgt ttgttgaact aatgggtgct ttagttgaag aataaaagac 2401 cacattaaaa aatgtggtct tttgtgtttt tttaaaggat ttgagcgtag cgaaaaatcc 2461 ttttctttct tatcttgata ataagggtaa ctattgccgg cgaggctagt tacccttaag 2521 ttattggtat gactggtttt aagcgcaaaa aaagttgctt tttcgtacct attaatgtat 2581 cgttttaaat gaatagtaaa aaacatacat agaaagggga aaaagcaact ttttttattg 2641 tcatagtttg tgaaaactaa gttgttttta tgtgttataa catggaaaag tatactgaga 2701 aaaaacaaag aaatcaagta tttcagaaat ttattaaacg tcatattgga gagaatcaaa 2761 tggatttagt tgaagattgc aatacatttc tgtcttttgt agctgataaa actttagaaa 2821 aacagaaatt atataaagct aattcttgta aaaatcgatt ttgtcctgtc tgtgcttgga 2881 gaaaagctag aaaagatgca ttgggtttat ctttgatgat gcaatatatt aagcagcaag 2941 agaaaaagga gtttatcttt ttaactttga ctacacctaa tgtaatgagt gatgaattag 3001 aaaatgaaat aaaacgttat aataattctt ttagaaaact tataaagaga aaaaaagtag 3061 gtagtgttat aaagggatat gttcgtaagt tagagattac atataataaa aaaagagatg 3121 attataatcc tcattttcat gtgttaattg cagtaaataa atcgtatttc acagataaaa 3181 gatattatat tagccaacaa gaatggttag atttatggcg tgatgtaacg ggcatttcag 3241 aaataacaca agttcaagtt caaaaaataa gacaaaataa taataaagaa ttatatgaaa 3301 tggctaagta ttctggtaaa gatagtgatt atttaataaa tcaaaaagtc tttgatgcat 3361 tttataaatc acttaaaggt aaacaggtat tagtttattc aggattattt aaagaggcta 3421 aaaagaaatt aaaaaatggg gatttagatt acttaaaaga aattgatcca accgaatata 3481 tctatcaaat tttttatatt tggaaacaaa aagagtattt agctagtgaa ctttatgact 3541 taacagaaca agaaaaaaga gaaattaatc acaaaatgat agacgaaatc gaggaagaac 3601 aataacaaaa tataagtgct aacagtcgtc tgcaagttta gttaagggtt cttctcaacg 3661 cacaataaat tttctcggca taaatgcgtg gtctaatttt tatttttaat aaccttgata 3721 gcaaaaaatg ccattccaat acaaaaccac atacctataa tcgataacca cataacagtc 3781 ataaaaccac tcctttttaa caaactttat cacaagaaat attttggcat tctacgacta 3841 taacttaaat ttatattttt tactttataa tatataattg attatagaat aatgttgctc 3901 atatcgtttg ccaacatcta gtactcaaat tacactatgt tacacttggt aatattaacc 3961 gaacttcccc tgtccaaatt agtttttacg gttcctggcc ttttgctggc cttttgctca 4021 catgttcttt cctgcgttgt cgacagagtt tgcaaaatat acaggggatt atatataatg 4081 gaaacgagac cattggtctc agttttagag ctagaaatag caagttaaaa taaggctagt 4141 ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt ttttgagatc tgtccatacc 4201 catggtctag aatgctcgag tcagaaaaat atacctgtat cttttttcaa aagcaaacat 4261 gctttgggta aacatgtagg tattaacgtc aatgcgacaa tagtagcatt gattaaatga 4321 gaattagtaa gtgttttact tactaaattt tatttaacct aaaaatgaac cacctggatg 4381 tgtgggatta aaaagtgaag agaggaggac gtatcacatg gataagaaat actcaatagg 4441 cttagatatc ggcacaaata gcgtcggatg ggcggtgatc actgatgaat ataaggttcc 4501 gtctaaaaag ttcaaggttc tgggaaatac agaccgccac agtatcaaaa aaaatcttat 4561 aggggctctt ttatttgaca gtggagagac agcggaagcg actcgtctca aacggacagc 4621 tcgtagaagg tatacacgtc ggaagaatcg tatttgttat ctacaggaga ttttttcaaa 4681 tgagatggcg aaagtagatg atagtttctt tcatcgactt gaagagtctt ttttggtgga 4741 agaagacaag aagcatgaac gtcatcctat ttttggaaat atagtagatg aagttgctta 4801 tcatgagaaa tatccaacta tctatcatct gcgaaaaaaa ttggtagatt ctactgataa 4861 agcggatttg cgcttaatct atttggcctt agcgcatatg attaagtttc gtggtcattt 4921 tttgattgag ggagatttaa atcctgataa tagtgatgtg gacaaactat ttatccagtt 4981 ggtacaaacc tacaatcaat tatttgaaga aaaccctatt aacgcaagtg gagtagatgc 5041 taaagcgatt ctttctgcac gattgagtaa atcaagacga ttagaaaatc tcattgctca 5101 gctccccggt gagaagaaaa atggcttatt tgggaatctc attgctttgt cattgggttt 5161 gacccctaat tttaaatcaa attttgattt ggcagaagat gctaaattac agctttcaaa 5221 agatacttac gatgatgatt tagataattt attggcgcaa attggagatc aatatgctga 5281 tttgtttttg gcagctaaga atttatcaga tgctatttta ctttcagata tcctaagagt 5341 aaatactgaa ataactaagg ctcccctatc agcttcaatg attaaacgct acgatgaaca 5401 tcatcaagac ttgactcttt taaaagcttt agttcgacaa caacttccag aaaagtataa 5461 agaaatcttt tttgatcaat caaaaaacgg atatgcaggt tatattgatg ggggagctag 5521 ccaagaagaa ttttataaat ttatcaaacc aattttagaa aaaatggatg gtactgagga 5581 attattggtg aaactaaatc gtgaagattt gctgcgcaag caacggacct ttgacaacgg 5641 ctctattccc catcaaattc acttgggtga gctgcatgct attttgagaa gacaagaaga 5701 cttttatcca tttttaaaag acaatcgtga gaagattgaa aaaatcttga cttttcgaat 5761 tccttattat gttggtccat tggcgcgtgg caatagtcgt tttgcatgga tgactcggaa 5821 gtctgaagaa acaattaccc catggaattt tgaagaggtt gtcgataaag gtgcttcagc 5881 tcaatcattt attgaacgca tgacaaactt tgataaaaat cttccaaatg aaaaagtact 5941 accaaaacat agtttgcttt atgagtattt tacggtttat aacgaattga caaaggtcaa 6001 atatgttact gaaggaatgc gaaaaccagc atttctttca ggtgaacaga agaaagccat 6061 tgttgattta ctcttcaaaa caaatcgaaa agtaaccgtt aagcaattaa aagaagatta 6121 tttcaaaaaa atagaatgtt ttgatagtgt tgaaatttca ggagttgaag atagatttaa 6181 tgcttcatta ggtacctacc atgatttgct aaaaattatt aaagataaag attttttgga 6241 taatgaagaa aatgaagata tcttagagga tattgtttta acattgacct tatttgaaga 6301 tagagagatg attgaggaaa gacttaaaac atatgctcac ctctttgatg ataaggtgat 6361 gaaacagctt aaacgtcgcc gttatactgg ttggggacgt ttgtctcgaa aattgattaa 6421 tggtattagg gataagcaat ctggcaaaac aatattagat tttttgaaat cagatggttt 6481 tgccaatcgc aattttatgc agctgatcca tgatgatagt ttgacattta aagaagacat 6541 tcaaaaagca caagtgtctg gacaaggcga tagtttacat gaacatattg caaatttagc 6601 tggtagccct gctattaaaa aaggtatttt acagactgta aaagttgttg atgaattggt 6661 caaagtaatg gggcggcata agccagaaaa tatcgttatt gaaatggcac gtgaaaatca 6721 gacaactcaa aagggccaga aaaattcgcg agagcgtatg aaacgaatcg aagaaggtat 6781 caaagaatta ggaagtcaga ttcttaaaga gcatcctgtt gaaaatactc aattgcaaaa 6841 tgaaaagctc tatctctatt atctccaaaa tggaagagac atgtatgtgg accaagaatt 6901 agatattaat cgtttaagtg attatgatgt cgatcacatt gttccacaaa gtttccttaa 6961 agacgattca atagacaata aggtcttaac gcgttctgat aaaaatcgtg gtaaatcgga 7021 taacgttcca agtgaagaag tagtcaaaaa gatgaaaaac tattggagac aacttctaaa 7081 cgccaagtta atcactcaac gtaagtttga taatttaacg aaagctgaac gtggaggttt 7141 gagtgaactt gataaagctg gttttatcaa acgccaattg gttgaaactc gccaaatcac 7201 taagcatgtg gcacaaattt tggatagtcg catgaatact aaatacgatg aaaatgataa 7261 acttattcga gaggttaaag tgattacctt aaaatctaaa ttagtttctg acttccgaaa 7321 agattttcaa ttctataaag tacgtgagat taacaattac catcatgccc atgatgcgta 7381 tctaaatgcc gtcgttggaa ctgctttgat taagaaatat ccaaaacttg aatcggagtt 7441 tgtctatggt gattataaag tttatgatgt tcgtaaaatg attgctaagt ctgagcaaga 7501 aataggcaaa gcaaccgcaa aatatttctt ttactctaat atcatgaact ttttcaaaac 7561 agaaattaca cttgcaaatg gagagattcg caaacgccct ctaatcgaaa ctaatgggga 7621 aactggagaa attgtctggg ataaagggcg agattttgcc acagtgcgca aagtattgtc 7681 catgccccaa gtcaatattg tcaagaaaac agaagtacag acaggcggat tctccaagga 7741 gtcaatttta ccaaaaagaa attcggacaa gcttattgct cgtaaaaaag actgggatcc 7801 aaaaaaatat ggtggttttg atagtccaac ggtagcttat tcagtcctag tggttgctaa 7861 ggtggaaaaa gggaaatcga agaagttaaa atccgttaaa gagttactag ggatcacaat 7921 tatggaaaga agttcctttg aaaaaaatcc gattgacttt ttagaagcta aaggatataa 7981 ggaagttaaa aaagacttaa tcattaaact acctaaatat agtctttttg agttagaaaa 8041 cggtcgtaaa cggatgctgg ctagtgccgg agaattacaa aaaggaaatg agctggctct 8101 gccaagcaaa tatgtgaatt ttttatattt agctagtcat tatgaaaagt tgaagggtag 8161 tccagaagat aacgaacaaa aacaattgtt tgtggagcag cataagcatt atttagatga 8221 gattattgag caaatcagtg aattttctaa gcgtgttatt ttagcagatg ccaatttaga 8281 taaagttctt agtgcatata acaaacatag agacaaacca atacgtgaac aagcagaaaa 8341 tattattcat ttatttacgt tgacgaatct tggagctccc gctgctttta aatattttga 8401 tacaacaatt gatcgtaaac gatatacgtc tacaaaagag gttttagatg ccactcttat 8461 ccatcaatcc atcactggtc tttatgaaac acgcattgat ttgagtcagc taggaggtga 8521 ctaatcgacc tgcagccaag cttgggcttt tcagcctgat acagattaaa tcagaacgca 8581 gaagcggtct gataaaacag aatttgcctg gcggcagtag cgcggtggtc ccacctgacc 8641 ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg tagtgtggcg tctccccttg 8701 cgagagtagg gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc 8761 tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga 8821 gcggatttga acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa 8881 actgccaggc atcaaattaa gcagaaggcc atcctgacgg atggcctttt tgcgtttcta 8941 caaactcttt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgcccatcgt 9001 atcgatgata agctgtcaaa catgagaatt aaatcaatct aaagtatata tgagtaaact 9061 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt 9121 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta 9181 ccatctggcc ccagtgctgc aatgataccg cgtgacccac gctcaccggc tccagattta 9241 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc 9301 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat 9361 agtttgcgca acgttgttgc cattgctact ggcatcgtag tgtcacgctc gtcgtttggt 9421 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg 9481 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca 9541 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta 9601 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg 9661 cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact 9721 ttaaaagtgc tcatcattgg aaaacgctct tcggggcgaa aactctcaag gatcttaccg 9781 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt 9841 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga 9901 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc 9961 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa 10021 caaatagggg ttccgcgcac atttccccga aaagtgccac ct //
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