质粒名称: | pCRISPomyces-2 |
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出品公司: | Addgene |
目录编号: | 61737 |
存储实验室: | Huimin Zhao |
载体骨架基本信息 |
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载体名称: | pKC1139 |
载体出品公司: | -- |
载体抗性: | 安普霉素 |
空载体大小: | -- |
完整质粒大小: | 10995 bp |
载体修饰: | 突变G8694C从pSG5-rep中去除BbsI位点 |
载体类型: | 细菌表达,CRISPR |
筛选标记: | -- |
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: | sSpCas9,Streptomyces codon-optimized Cas9 |
物种来源: | Synthetic |
插入大小: | 4107 bp |
基因突变: | -- |
GenBank编号 | -- |
启动子 | rpsL(XC)-BbsI |
标签/融合蛋白: | -- |
克隆方法: | -- |
5' 克隆位点: | -- |
3' 克隆位点: | -- |
5' 测序引物 | -- |
3' 测序引物 | -- |
文献引用方法 |
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材料和方法部分: | pCRISPomyces-2 was a gift from Huimin Zhao (Addgene plasmid # 61737 ; http://n2t.net/addgene:61737 ; RRID:Addgene_61737) |
参考文献部分: | High-Efficiency Multiplex Genome Editing of Streptomyces Species Using an Engineered CRISPR/Cas System. Cobb RE, Wang Y, Zhao H. ACS Synth Biol. 2014 Dec 8. 10.1021/sb500351f PubMed 25458909 |
买家导航
pCRISPomyces2载体基本信息
pCRISPomyces2载体图谱质粒图谱和多克隆位点信息
pCRISPomyces2载体简介
pCRISPomyces2载体序列
LOCUS pCRISPomyces-2 10994 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION Streptomyces expression of codon-optimized Cas9 and custom gRNA. ACCESSION . VERSION . KEYWORDS pCRISPomyces-2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10994) AUTHORS Cobb RE, Wang Y, Zhao H TITLE High-Efficiency Multiplex Genome Editing of Streptomyces Species Using an Engineered CRISPR/Cas System. JOURNAL ACS Synth Biol. 2014 Dec 8. PUBMED 25458909 REFERENCE 2 (bases 1 to 10994) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10994 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(153..1595) /codon_start=1 /gene="rep" /product="replication initiator protein from the Streptomyces ghanaensis plasmid pSG5" /label=pSG5 Rep /translation="MWKPGEATWGNTCRCNNVHTCPWCMSRILAVRGSNVQLAADGLAD AGYGLHLGTNTLRHFERMAFGTVRKGMRHGLVAVLHDGWKGAYGSSGRRWRTMRDDFGI IGYERAFEDTFGWGSGWHLHWHTLWVTREVLGPDAQAAFRDALAGAWAAGVESAGGYTV SETCDRPGCSCEGKGHGTDVRPLNGADAADGDAGKQARYLYKDGDKTKGGVAKIGLELA GQNFKAGRGDDRMGPLDLGDAAAAELQRLRRPGPFVEKYREREFGVFQVRKHYRSQNLN RLIKELGIQQDVRTEEEITDDTEGLVAIAVIPAYIWYRYIARVAGRRLDLIKVAETYGL PGVRRLVESWGLVWGKDVLDPPAPEAPAAPGDLDADQMRFEVMSEEEAAFREARRKANE ARTEELAASLDRVRQPKKEAIRPTISLRKRLKPKPVTVDVKTPPPGAASPVCRRCKGKL APVLQPWGQHPGDCLRVDTAVA" oriT 2762..2871 /note="incP origin of transfer" promoter 3093..3394 /label=rpsL promoter /note="promoter for the Xylanimonas cellulosilytica ribosomal protein S12 gene" CDS 3395..7501 /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" promoter 7701..7985 /label=gapdh promoter /note="glyceraldehyde-3-phosphate dehydrogenase promoter from Eggerthella lenta" CDS complement(7994..8290) /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSG" primer_bind 8140..8162 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 8154..8171 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 8155..8171 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 8181..8200 /label=T7 /note="T7 promoter, forward primer" promoter 8181..8199 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(8247..8267) /label=T3 /note="T3 promoter, forward primer" promoter complement(8247..8265) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(8286..8302) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(8286..8302) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(8299..8321) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 8310..8326 /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(8334..8364) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8379..8400 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." misc_RNA 8443..8518 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" terminator 8540..8574 /label=lambda t0 terminator /note="minimal transcription terminator from phage lambda (Scholtissek and Grosse, 1987)" rep_origin complement(8616..9204) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(8696..8715) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 9408..10208 /codon_start=1 /gene="aac(3)IV" /product="aminoglycoside 3-N-acetyltransferase type IV " /label=ApmR /note="confers resistance to apramycin (Paget and Davies, 1996)" /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG" ORIGIN 1 cggacgctcg gactgggcgc gggattccag cagtaaccca ggtccgccgg ccacctcacg 61 gcaggcagac cctcggcttt cgcgccggga ccgccatgag accgccaccc ggatgtccgg 121 ggtggcggtc tcatggcggt ccctcagcgg ccctacgcga ccgctgtgtc gacgcggagg 181 cagtctccgg ggtgctggcc ccagggctgg agtaccgggg cgagcttgcc cttgcagcgg 241 cggcacacgg gcgacgcggc gccgggcggg ggagtcttga cgtcgacggt gaccggcttc 301 ggcttcaggc gcttccgaag gctgatcgtc gggcggatcg cctccttctt cggctgacgc 361 actcggtcga gcgaagcggc cagctcctcc gtgcgggcct cgttggcctt ccggcgcgcc 421 tcgcggaaag cagcttcctc ctcggacatg acctcgaacc tcatctggtc agcgtcaagg 481 tcgcccggcg cggccggcgc ttccggggcg ggcgggtcca ggacgtcctt gccccacacc 541 aggccccagg actcgacgag ccgccggacg cccggtaggc cgtacgtctc ggcgaccttg 601 atgaggtcga ggcgacgtcc ggcgacgcgg gcgatgtatc ggtaccagat gtaggccggg 661 atgaccgcga tggcgaccag gccctcggtg tcgtcggtga tctcctcctc ggtgcggacg 721 tcctgctgga tgccgagttc cttgatcagc cggttcaggt tctgcgaccg gtagtgcttg 781 cggacctgga acacgccgaa ctcgcgctcg cggtacttct cgacgaacgg gccgggccga 841 cgaagccgct gcagctcggc ggccgccgcg tcgcccaggt cgagcggtcc catgcggtcg 901 tcgccgcgac cggccttgaa gttctgtccg gccagctcca ggccgatctt ggcgacgccg 961 cccttggtct tgtcgccgtc cttgtagagg tagcgggcct gcttgcccgc atcgccgtca 1021 gcggcgtccg cgccgttgag tgggcgcacg tcggtgccgt ggcccttgcc ctcacaggag 1081 caaccgggcc ggtcgcacgt ctcgctgacg gtgtagccgc ccgcggattc gaccccggcg 1141 gcccaggctc cggcgagtgc gtcgcggaac gcggcctggg cgtccgggcc gagcacctcg 1201 cgggtgaccc agagcgtgtg ccagtgcagg tgccagccgg agccccagcc gaaggtgtcc 1261 tcgaaggccc gctcgtagcc gatgatcccg aagtcgtcgc gcatcgtgcg ccagcggcgg 1321 ccggacgagc cgtacgcgcc cttccagccg tcgtgcaaga ccgcgaccag gccgtgccgc 1381 attcccttgc ggacggtgcc gaacgccatg cgctcgaagt ggcgcaacgt gttcgtgcca 1441 aggtgcagcc cgtacccggc gtccgcgaga ccgtcggcgg cgagctgcac gttcgagccc 1501 cgtacggcca ggatgcggct catgcaccac gggcaggtgt ggacgttgtt gcagcggcac 1561 gtgttgcccc acgtcgcctc gcccggcttc cacatcagct cggccgtccc ggcagtgagc 1621 cgggtcccgc agcccttgaa cgcctcgttc agcgacaccg tctggtgccg gtcccgccgg 1681 gcgaaccgct cgtcgcgcgg gtcctcccgc ccggctgtcg cggcaccctc gtttggggta 1741 gaacccgttc cagttacagc gctctgacct gcagtggacg gagattttcc cttactacta 1801 aagcccgcgt ccggattacc cgctgtagtc gtgcttgcta cgctgcgtga ctggtccgca 1861 atgagacgct ttgcgcgctt tcggcaggcg tccgagcagt agattttggg gcgcttcccg 1921 gggatgtgga cgatcggggt gccgcagtgg cacttcggtc cggcggggcg cggtggtgtc 1981 gacgcgctgt tctctcgtac gctcgtcaca gagcaaacgt cctcactcgg catgctgcgc 2041 cggttcgggg gcggcgagcc cgggaggcca atcccgggct cgtgccattt ctgggtcctg 2101 ttgatcctgg cattggtgtg gccgttcatt gcccctgctc gctcctgacg cgccgataga 2161 cgtccgatac gcccggtgct ggtgggattt gataggtcgg aagaagcccc gcccgggcct 2221 gggcggggct tcctgtgcgt caggacctcc tcgtcgtgag cctcttcggc ctatggacgg 2281 agtgacctcg tgatccgtta cagccgcgcg cgctcgcgta gagcggtctc atcagttcca 2341 cgaacggtcc tcttcgcaga tcagggcgtt ggggcggagt ctcaccaagg actacgtctg 2401 ctggcgattt ccgttacacc ccgggcggtg gccggcgcac acgcgcgccc gcgttgggca 2461 gtgcagaaag tgcagaaacc taggcgctga tggtccaggt ccacggttcg tcgtcggcgg 2521 cggcgcgggc ggcggcgtcg gccagggcgc gggcgagacc ggctacggcg ggcttgatgc 2581 gccggttgcg ggcgaccttg agcagctagt atgcaggtcg acggatcttt tccgctgcat 2641 aaccctgctt cggggtcatt atagcgattt tttcggtata tccatccttt ttcgcacgat 2701 atacaggatt ttgccaaagg gttcgtgtag actttccttg gtgtatccaa cggcgtcagc 2761 cgggcaggat aggtgaagta ggcccacccg cgagcgggtg ttccttcttc actgtccctt 2821 attcgcacct ggcggtgctc aacgggaatc ctgctctgcg aggctggccg gctaccgccg 2881 gcgtaacaga tgagggcaag cggatggctg atgaaaccaa gccaaccagg aagggcagcc 2941 cacctatcaa ggtgtactgc cttccagacg aacgaagagc gattgaggaa aaggcggcgg 3001 cggccggcat gagcctgtcg gcctacctgc tggccgtcgg ccagggctac aaaatcacgg 3061 gcgtcgtgga ctatgagcac gtccgcgagc tggccctgca ggcggaagtc aggtagacac 3121 gacttccgct agtccttgca aggtctgctg acgtgaggcg gggcggtcgt ttttgaccgc 3181 cctgccttcg tcatgtaggc tcgctcgctg tgcctggcgt gtcatcagac gcccaggtcc 3241 cggtgccgtg aggcccgggc catcgagccg gtggtacgtg gctgcggtcc ccttgtgagg 3301 gctgcgcgcc gtgtgctgtc cggcgcgcac agccttgaat ccacccgcgg gggccggccg 3361 gtctccgtga gctcgagtag acgacggaga cgtaatggac aagaagtaca gcatcggcct 3421 ggacatcggc accaacagcg tgggctgggc ggtcatcacc gacgagtaca aggtcccctc 3481 caagaagttc aaggtcctgg gcaacaccga ccggcactcg atcaagaaga acctgatcgg 3541 cgccctgctc ttcgacagcg gcgaaaccgc cgaggcgacc cgcctgaagc ggaccgcccg 3601 tcgccgctac acccggcgca agaaccgcat ctgctacctg caggagatct tctccaacga 3661 gatggccaag gtcgacgact cgttcttcca ccggctcgag gagagcttcc tggtggagga 3721 ggacaagaag cacgagcgcc acccgatctt cggcaacatc gtcgacgagg tggcctacca 3781 cgagaagtac cccaccatct accacctccg caagaagctg gtcgactcga ccgacaaggc 3841 ggacctgcgg ctcatctacc tggccctcgc gcacatgatc aagttccgcg gccacttcct 3901 catcgagggc gacctgaacc cggacaactc cgacgtcgac aagctcttca tccagctggt 3961 gcagacctac aaccagctgt tcgaggagaa ccccatcaac gccagcggcg tcgacgccaa 4021 ggcgatcctc tccgcgcgcc tgagcaagtc ccggcgcctg gagaacctca tcgcccagct 4081 gccgggcgag aagaagaacg gcctcttcgg caacctgatc gcgctgtcgc tcggcctgac 4141 ccccaacttc aagagcaact tcgacctggc cgaggacgcg aagctccagc tgtccaagga 4201 cacctacgac gacgacctgg acaacctgct cgcccagatc ggcgaccagt acgcggacct 4261 cttcctggcc gcgaagaacc tctcggacgc catcctgctc agcgacatcc tgcgggtcaa 4321 caccgagatc accaaggccc cgctgtcggc gagcatgatc aagcggtacg acgagcacca 4381 ccaggacctg accctgctca aggccctcgt gcgccagcag ctgcccgaga agtacaagga 4441 gatcttcttc gaccagtcca agaacggcta cgccggctac atcgacggcg gcgcgtcgca 4501 ggaggagttc tacaagttca tcaagccgat cctggagaag atggacggca ccgaggagct 4561 gctcgtcaag ctgaaccgcg aggacctgct ccgcaagcag cggaccttcg acaacggctc 4621 catcccgcac cagatccacc tgggcgagct ccacgccatc ctccggcgcc aggaggactt 4681 ctaccccttc ctgaaggaca accgcgagaa gatcgagaag atcctgacct tccggatccc 4741 gtactacgtc ggccccctgg cccgcggcaa ctcccggttc gcgtggatga cccggaagtc 4801 ggaggaaacc atcaccccgt ggaacttcga ggaggtcgtg gacaagggcg cctccgcgca 4861 gtcgttcatc gagcgcatga ccaacttcga caagaacctc ccgaacgaga aggtcctgcc 4921 caagcacagc ctgctctacg agtacttcac cgtgtacaac gagctgacca aggtcaagta 4981 cgtgaccgag ggcatgcgga agccggcctt cctgtccggc gagcagaaga aggcgatcgt 5041 cgacctgctc ttcaagacca accgcaaggt caccgtgaag cagctgaagg aggactactt 5101 caagaagatc gagtgcttcg actccgtcga gatctcgggc gtggaggacc gcttcaacgc 5161 ctccctgggc acctaccacg acctgctcaa gatcatcaag gacaaggact tcctcgacaa 5221 cgaggagaac gaggacatcc tggaggacat cgtcctcacc ctgaccctct tcgaggaccg 5281 cgagatgatc gaggagcggc tcaagaccta cgcccacctg ttcgacgaca aggtgatgaa 5341 gcagctgaag cggcgccggt acaccggctg gggccgcctc tcccggaagc tgatcaacgg 5401 catccgggac aagcagagcg gcaagaccat cctggacttc ctcaagtccg acggcttcgc 5461 caaccgcaac ttcatgcagc tcatccacga cgactcgctg accttcaagg aggacatcca 5521 gaaggcccag gtgtccggcc agggcgacag cctccacgag cacatcgcca acctggcggg 5581 ctccccggcg atcaagaagg gcatcctcca gaccgtcaag gtcgtggacg agctggtcaa 5641 ggtgatgggc cgccacaagc ccgagaacat cgtgatcgag atggcccggg agaaccagac 5701 cacccagaag ggccagaaga actcccgcga gcggatgaag cgcatcgagg agggcatcaa 5761 ggagctcggc tcgcagatcc tgaaggagca cccggtcgag aacacccagc tccagaacga 5821 gaagctgtac ctctactacc tgcagaacgg ccgcgacatg tacgtggacc aggagctcga 5881 catcaaccgg ctgagcgact acgacgtcga ccacatcgtg ccgcagtcct tcctgaagga 5941 cgactcgatc gacaacaagg tcctgacccg ctccgacaag aaccggggca agtccgacaa 6001 cgtgccctcg gaggaggtcg tgaagaagat gaagaactac tggcgccagc tgctcaacgc 6061 caagctcatc acccagcgca agttcgacaa cctgaccaag gccgagcggg gcggcctgtc 6121 ggagctcgac aaggcgggct tcatcaagcg ccagctcgtc gaaacccggc agatcaccaa 6181 gcacgtggcc cagatcctgg acagccggat gaacaccaag tacgacgaga acgacaagct 6241 gatccgcgag gtcaaggtga tcaccctcaa gagcaagctg gtgtccgact tccgcaagga 6301 cttccagttc tacaaggtcc gggagatcaa caactaccac cacgcccacg acgcgtacct 6361 gaacgccgtc gtgggcaccg cgctgatcaa gaagtacccg aagctggagt ccgagttcgt 6421 ctacggcgac tacaaggtct acgacgtgcg caagatgatc gccaagtcgg agcaggagat 6481 cggcaaggcc accgcgaagt acttcttcta cagcaacatc atgaacttct tcaagaccga 6541 gatcaccctg gccaacggcg agatccgcaa gcggcccctg atcgaaacca acggcgaaac 6601 cggcgagatc gtctgggaca agggccgcga cttcgccacc gtccggaagg tgctgtccat 6661 gccgcaggtc aacatcgtca agaaaaccga ggtgcagacc ggcggcttca gcaaggagtc 6721 catcctcccc aagcgcaact cggacaagct gatcgcccgg aagaaggact gggacccgaa 6781 gaagtacggc ggcttcgaca gccccaccgt cgcctactcc gtgctggtcg tggcgaaggt 6841 cgagaagggc aagagcaaga agctgaagtc cgtgaaggag ctgctcggca tcaccatcat 6901 ggagcgctcc tcgttcgaga agaacccgat cgacttcctg gaggccaagg gctacaagga 6961 ggtcaagaag gacctcatca tcaagctgcc caagtactcg ctgttcgagc tcgagaacgg 7021 ccgcaagcgg atgctcgcca gcgcgggcga gctgcagaag ggcaacgagc tggccctccc 7081 gtccaagtac gtcaacttcc tgtacctcgc gtcccactac gagaagctga agggctcgcc 7141 cgaggacaac gagcagaagc agctcttcgt ggagcagcac aagcactacc tggacgagat 7201 catcgagcag atctcggagt tcagcaagcg ggtcatcctg gccgacgcga acctcgacaa 7261 ggtgctgtcc gcctacaaca agcaccgcga caagccgatc cgggagcagg cggagaacat 7321 catccacctg ttcaccctca ccaacctggg cgcccccgcc gcgttcaagt acttcgacac 7381 caccatcgac cgcaagcggt acaccagcac caaggaggtc ctcgacgcga ccctgatcca 7441 ccagtccatc accggcctgt acgaaacccg catcgacctc tcccagctcg gcggcgactg 7501 agaattcaga tctacgcgtt ccccgcaaaa gcggcctttg actccctgca agcctcagcg 7561 accgaatata tcggttatgc gtgggcgatg gttgttgtca ttgtcggcgc aactatcggt 7621 atcaagctgt ttaagaaatt cacctcgaaa gcaagctgat aaaccgatac aattaaaggc 7681 tccttttgga gccttttttt gctgctcctt cggtcggacg tgcgtctacg ggcaccttac 7741 cgcagccgtc ggctgtgcga cacggacgga tcgggcgaac tggccgatgc tgggagaagc 7801 gcgctgctgt acggcgcgca ccgggtgcgg agcccctcgg cgagcggtgt gaaacttctg 7861 tgaatggcct gttcggttgc tttttttata cggctgccag ataaggcttg cagcatctgg 7921 gcggctaccg ctatgatcgg ggcgttcctg caattcttag tgcgagtatc tgaaagggga 7981 tacgcaagtc ttctcagccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact 8041 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat 8101 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa 8161 cgacggccag tgagcgcgcg taatacgact cactataggg cgaattgggt accgggcccc 8221 ccctcgaggt cctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa 8281 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata 8341 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta 8401 attgcgttgc gctcactgcc cgctttccac cggtgaagac atgttttaga gctagaaata 8461 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt 8521 tttactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttatctagag 8581 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 8641 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 8701 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 8761 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc 8821 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgacccc 8881 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 8941 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 9001 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca 9061 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 9121 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 9181 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 9241 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc 9301 acctagatcc ttttggttca tgtgcagctc catcagcaaa aggggatgat aagtttatca 9361 ccaccgacta tttgcaacag tgccgttgat cgtgctatga tcgactgatg tcatcagcgg 9421 tggagtgcaa tgtcgtgcaa tacgaatggc gaaaagccga gctcatcggt cagcttctca 9481 accttggggt tacccccggc ggtgtgctgc tggtccacag ctccttccgt agcgtccggc 9541 ccctcgaaga tgggccactt ggactgatcg aggccctgcg tgctgcgctg ggtccgggag 9601 ggacgctcgt catgccctcg tggtcaggtc tggacgacga gccgttcgat cctgccacgt 9661 cgcccgttac accggacctt ggagttgtct ctgacacatt ctggcgcctg ccaaatgtaa 9721 agcgcagcgc ccatccattt gcctttgcgg cagcggggcc acaggcagag cagatcatct 9781 ctgatccatt gcccctgcca cctcactcgc ctgcaagccc ggtcgcccgt gtccatgaac 9841 tcgatgggca ggtacttctc ctcggcgtgg gacacgatgc caacacgacg ctgcatcttg 9901 ccgagttgat ggcaaaggtt ccctatgggg tgccgagaca ctgcaccatt cttcaggatg 9961 gcaagttggt acgcgtcgat tatctcgaga atgaccactg ctgtgagcgc tttgccttgg 10021 cggacaggtg gctcaaggag aagagccttc agaaggaagg tccagtcggt catgcctttg 10081 ctcggttgat ccgctcccgc gacattgtgg cgacagccct gggtcaactg ggccgagatc 10141 cgttgatctt cctgcatccg ccagaggcgg gatgcgaaga atgcgatgcc gctcgccagt 10201 cgattggctg agctcatgag cggagaacga gatgacgttg gaggggcaag gtcgcgctga 10261 ttgctggggc aacacgtgga gcggatcggg gattgtcttt cttcagctcg ctgatgatat 10321 gctgacgctc aatgccgttt ggcctccgac taacgaaaat cccgcatttg gacggctgat 10381 ccgattggca cggcggacgg cgaatggcgg agcagacgct cgtccggggg caatgagata 10441 tgaaaaagcc tgaactcacc gcgacgtatc gatgtcgagg ttcctcaggg gagccacccc 10501 agagaagccc tcggagctga gcggagctat ttccaaagcc atgccagcta gagacagtgc 10561 acaccgccaa gcatctgcaa aaccctcgcc tggagggaaa gtgcaatgta cgtacgctgg 10621 tttccctcca gaaagggatt ctgcggcttc tcaactttgg aagaagaggc ggaacgaact 10681 gctgtttcag cctactcatg tgaggaggct ggtcctttac cctgatgccc gagggcatca 10741 gggtaaagga ccagcctcgc caactaggag cgctaccaag gcgaacacaa cccaggactt 10801 ggtagaaccc ttcgccggag aagttcagac tcatcggcat gagcacccct cgccgcgcac 10861 tcggcaccgg tccgtcgacc accaccagga cgtcgttgtc gacgtcggcc ccgcggctcc 10921 tgcccgccga acgcgtcgtc gtcgacggcc tggtgctcat cgacgagcac ccggagccag 10981 gtgaaaagcg ccgg //
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