质粒名称: | pCS2-nCas9n |
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出品公司: | 爱迪基因 |
目录编号: | 47929 |
存储实验室: | Wenbiao Chen |
载体骨架基本信息 |
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载体名称: | pCS2+ |
载体出品公司: | -- |
载体抗性: | 氨苄青霉素 |
空载体大小: | 4100 bp |
完整质粒大小: | 8300 bp |
载体修饰: | -- |
载体类型: | 哺乳动物表达,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: | nls-zcas9-nls |
物种来源: | Synthetic |
插入大小: | 4200 bp |
基因突变: | -- |
GenBank编号 | -- |
启动子 | CMV |
标签/融合蛋白: | -- |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | BamHI(未破坏) |
3' 克隆位点: | XbaI(未破坏) |
5' 测序引物 | SP6 |
3' 测序引物 | M13 Rev |
文献引用方法 |
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材料和方法部分: | pCS2-nCas9n was a gift from Wenbiao Chen (Addgene plasmid # 47929 ; http://n2t.net/addgene:47929 ; RRID:Addgene_47929) |
参考文献部分: | Efficient multiplex biallelic zebrafish genome editing using a CRISPR nuclease system. Jao LE, Wente SR, Chen W. Proc Natl Acad Sci U S A. 2013 Aug 5. 10.1073/pnas.1308335110 PubMed 23918387 |
买家导航
pCS2nCas9n载体基本信息
pCS2nCas9n载体图谱质粒图谱和多克隆位点信息
pCS2nCas9n载体简介
pCS2nCas9n载体序列
LOCUS pCS2-nCas9n 8269 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION expression of an optimized Cas9 for genome-editing in zebrafish. ACCESSION . VERSION . KEYWORDS pCS2-nCas9n SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8269) AUTHORS Jao LE, Wente SR, Chen W TITLE Efficient multiplex biallelic zebrafish genome editing using a CRISPR nuclease system. JOURNAL Proc Natl Acad Sci U S A. 2013 Aug 5. PUBMED 23918387 REFERENCE 2 (bases 1 to 8269) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8269 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 30..1007 /label=CMV IE94 promoter /note="enhancer/promoter region of simian cytomegalovirus major immediate early transcription unit IE94" promoter 1042..1060 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 1042..1059 /label=SP6 /note="SP6 promoter, forward primer" regulatory 1106..1115 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1124..1144 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1151..5251 /codon_start=1 /product="Cas9 (Csn1) endonuclease from the Streptococcus pyogenes MGAS15252 Type II CRISPR/Cas system" /label=Cas9 /note="generates RNA-guided double strand breaks in DNA" /translation="MDKKYSIGLDIGTNSVGWAVITDDYKVPSKKFKVLGNTDRHSIKK NLIGALLFGSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLADSTDKADLRLIYLALAHMIK FRGHFLIEGDLNPDNSDVDKLFIQLVQIYNQLFEENPINASRVDAKAILSARLSKSRRL ENLIAQLPGEKRNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNSEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD SVEISGVEDRFNASLGAYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDRGMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGHSLHEQIANLAGSPAIKKGILQTVKIVDELVKVMGH KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFIKDDSIDNKVLTRSDKNRGKSDNVPS EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD ATLIHQSITGLYETRIDLSQLGGD" RBS 5164..5172 /label=Shine-Dalgarno sequence /note="full consensus sequence for ribosome-binding sites upstream of start codons in E. coli; complementary to a region in the 3' end of the 16S rRNA (Chen et al., 1994)" CDS 5267..5287 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" promoter complement(5306..5323) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" polyA_signal 5328..5462 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5352..5371) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 5406..5425 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(5575..5595) /label=T3 /note="T3 promoter, forward primer" promoter complement(5575..5593) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(5614..5630) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5614..5630) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5627..5649) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5638..5654 /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(5662..5692) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5707..5728 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5845..5862) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6016..6604) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6096..6115) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6775..7635) /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 7398..7417 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7636..7740) /gene="bla" /label=AmpR promoter rep_origin complement(7766..8221) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(7903..7924) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 8115..8134 /label=F1ori-R /note="F1 origin, reverse primer" ORIGIN 1 gggcctcttc gctattacgc cagtcgaccg ccaattcaat atggcgtata tggactcatg 61 ccaattcaat atggtggatc tggacctgtg ccaattcaat atggcgtata tggactcgtg 121 ccaattcaat atggtggatc tggaccccag ccaattcaat atggcggact tggcaccatg 181 ccaattcaat atggcggacc tggcactgtg ccaactgggg aggggtctac ttggcacggt 241 gccaagtttg aggaggggtc ttggccctgt gccaagtccg ccatattgaa ttggcatggt 301 gccaataatg gcggccatat tggctatatg ccaggatcaa tatataggca atatccaata 361 tggccctatg ccaatatggc tattggccag gttcaatact atgtattggc cctatgccat 421 atagtattcc atatatgggt tttcctattg acgtagatag cccctcccaa tgggcggtcc 481 catataccat atatggggct tcctaatacc gcccatagcc actcccccat tgacgtcaat 541 ggtctctata tatggtcttt cctattgacg tcatatgggc ggtcctattg acgtatatgg 601 cgcctccccc attgacgtca attacggtaa atggcccgcc tggctcaatg cccattgacg 661 tcaataggac cacccaccat tgacgtcaat gggatggctc attgcccatt catatccgtt 721 ctcacgcccc ctattgacgt caatgacggt aaatggccca cttggcagta catcaatatc 781 tattaatagt aacttggcaa gtacattact attggaagta cgccagggta cattggcagt 841 actcccattg acgtcaatgg cggtaaatgg cccgcgatgg ctgccaagta catccccatt 901 gacgtcaatg gggaggggca atgacgcaaa tgggcgttcc attgacgtaa atgggcggta 961 ggcgtgccta atgggaggtc tatataagca atgctcgttt agggaaccgc cattctgcct 1021 ggggacgtcg gagcaagctt gatttaggtg acactataga atacaagcta cttgttcttt 1081 ttgcaggatc cgatccaatg tcgacgccac catggcttct ccacctaaga agaagagaaa 1141 ggtgggaagc atggataaga agtatagcat cggcctggat attggaacta actccgtggg 1201 ttgggcagtg attacagacg actacaaggt ccctagcaag aaatttaagg tgctgggtaa 1261 caccgacagg cacagcatca agaaaaatct gattggagcc ctgctgttcg gttctggaga 1321 gactgccgaa gcaacacgcc tgaaaagaac agcaagaagg cgctatacca gaaggaagaa 1381 tagaatctgt tacctgcagg agattttctc taacgaaatg gctaaggtgg acgattcatt 1441 ctttcatagg ctggaggaaa gtttcctggt cgaggaagat aagaaacacg agcgccatcc 1501 tatctttgga aacattgtgg acgaggtcgc ctatcacgaa aaatacccaa ccatctatca 1561 tctgcgcaag aaactggctg actctactga taaagccgac ctgagactga tctatctggc 1621 tctggcccac atgattaagt tcaggggtca ttttctgatc gagggcgatc tgaaccccga 1681 caattccgat gtggacaagc tgttcatcca gctggtccag atttacaatc agctgtttga 1741 ggaaaaccct attaatgctt ccagagtgga cgcaaaagct atcctgtcag ccaggctgtc 1801 caagtcacgc agactggaga acctgattgc acagctgccc ggagaaaaga ggaacggtct 1861 gtttggaaat ctgatcgctc tgagtctggg cctgactcct aacttcaaaa gcaattttga 1921 tctggctgag gacgccaaac tgcagctgtc aaaggacaca tatgacgatg acctggataa 1981 cctgctggca cagatcggag atcagtacgc tgacctgttc ctggctgcca aaaatctgtc 2041 cgacgcaatc ctgctgtcag atattctgag agtgaacagc gagattacaa aagctcctct 2101 gagtgccagc atgatcaaga gatatgacga gcaccatcag gatctgaccc tgctgaaggc 2161 tctggtcagg cagcagctgc cagagaagta caaggaaatt ttctttgatc agtccaagaa 2221 cggctacgcc ggttatatcg acggaggcgc atcacaggag gaattctaca agtttatcaa 2281 acctattctg gagaagatgg acggaactga ggaactgctg gtgaaactga atagagagga 2341 cctgctgagg aagcagcgca catttgataa cggttccatc ccacaccaga ttcatctggg 2401 agagctgcac gctatcctga ggcgccagga agacttctac ccctttctga aagataaccg 2461 cgagaagatc gaaaaaattc tgaccttcag aatcccttac tatgtgggtc cactggctcg 2521 cggaaacagc agatttgcct ggatgactcg caaatccgag gaaaccatta ctccttggaa 2581 cttcgaggaa gtggtcgata agggcgcctc tgcacagtcc ttcatcgaga gaatgactaa 2641 ttttgacaaa aacctgccca atgagaaagt gctgcctaag cactccctgc tgtacgagta 2701 tttcactgtc tataacgaac tgacaaaggt gaaatacgtc accgagggca tgagaaagcc 2761 agccttcctg tcaggagagc agaagaaagc aatcgtggat ctgctgttta aaaccaacag 2821 gaaagtgact gtcaagcagc tgaaggagga ctacttcaag aaaattgaat gcttcgattc 2881 cgtggagatc agcggagtcg aagacagatt taacgcaagc ctgggcgctt accacgatct 2941 gctgaagatc attaaggata aagacttcct ggacaacgag gaaaatgagg atatcctgga 3001 agacattgtg ctgacactga ccctgtttga ggacagagga atgatcgagg aaagactgaa 3061 aacctatgct catctgttcg atgacaaggt gatgaaacag ctgaagagaa ggcgctacac 3121 tggctggggt agactgagca ggaagctgat caacggcatt agggataaac agtcaggaaa 3181 gacaatcctg gactttctga aaagtgatgg cttcgccaac cgcaatttta tgcagctgat 3241 tcacgatgac agtctgacct tcaaagagga catccagaag gctcaggtgt ctggacaggg 3301 ccactccctg catgagcaga ttgcaaacct ggctggaagc ccagccatca agaaaggcat 3361 tctgcagaca gtgaaaatcg tcgatgagct ggtgaaagtc atgggccata agcccgaaaa 3421 catcgtgatt gagatggctc gcgaaaatca gacaacccag aagggtcaga agaacagtag 3481 agagaggatg aaaagaatcg aggaaggcat taaggagctg ggtagccaga tcctgaaaga 3541 gcacccagtg gaaaacacac agctgcagaa tgagaagctg tatctgtact atctgcagaa 3601 tggaagagat atgtacgtgg accaggagct ggatattaac aggctgtctg attacgacgt 3661 ggatcatatc gtcccccaga gtttcatcaa agatgacagc attgacaaca aggtgctgac 3721 caggtccgac aaaaacagag gaaaatcaga taatgtccct agtgaggaag tggtcaagaa 3781 aatgaagaac tactggagac agctgctgaa tgccaaactg atcactcaga ggaagtttga 3841 taacctgaca aaagcagagc gcggtggact gtcagaactg gacaaagctg gattcatcaa 3901 gaggcagctg gtggaaacac gccagatcac taaacacgtc gcacagattc tggatagtcg 3961 catgaacaca aagtacgatg agaatgacaa actgatcaga gaagtgaagg tcattaccct 4021 gaagagtaaa ctggtcagcg actttaggaa agatttccag ttttataagg tccgcgagat 4081 taacaattat caccatgccc atgacgcata cctgaacgcc gtggtcggta ccgcactgat 4141 caagaaatac ccaaaactgg agagcgaatt cgtgtacgga gactataagg tgtacgatgt 4201 cagaaaaatg atcgccaagt ccgagcagga aattggaaaa gctactgcca agtatttctt 4261 ttactcaaac atcatgaatt tctttaagac agagatcacc ctggccaatg gagaaatccg 4321 caaaaggccc ctgattgaga caaacggaga gacaggcgaa atcgtgtggg acaaaggcag 4381 agattttgca accgtgagga aggtcctgag catgcctcaa gtgaatatcg tcaagaaaac 4441 tgaggtgcag acaggcggtt tctcaaaaga aagtattctg ccaaaacgca actctgataa 4501 gctgatcgct agaaagaaag actgggaccc taagaagtat ggaggctttg actctcccac 4561 tgtggcatac tccgtcctgg tggtcgctaa ggtggagaag ggcaaaagca agaaactgaa 4621 atctgtcaag gagctgctgg gtatcacaat tatggagaga agctctttcg agaagaaccc 4681 aatcgatttt ctggaggcca aaggttataa ggaagtgaag aaagacctga tcattaaact 4741 gcccaagtac agtctgtttg agctggaaaa cggcaggaaa cgcatgctgg caagcgctgg 4801 agagctgcag aaaggcaatg aactggccct gccttctaag tacgtgaact tcctgtatct 4861 ggcaagccac tacgagaagc tgaaaggatc tccagaggat aacgaacaga aacagctgtt 4921 tgtggagcag cacaagcatt atctggacga gatcattgaa cagattagcg agttctctaa 4981 aagagtgatc ctggccgacg caaatctgga taaggtcctg tctgcttaca acaaacacag 5041 agataagccc atcagggagc aggccgaaaa tatcattcat ctgttcactc tgacaaacct 5101 gggcgcacct gcagctttca agtacttcga cactacaatc gatagaaaga ggtacacctc 5161 cactaaggag gtgctggacg ctacactgat ccatcagagt attaccggcc tgtacgaaac 5221 aaggattgac ctgtctcagc tgggtggcga ctcaccggtg agatctccta agaagaagag 5281 aaaggtgtga ccgcggaatc tagaactata gtgagtcgta ttacgtagat ccagacatga 5341 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta 5401 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag 5461 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt 5521 tttaattcgc ggccgcggcg ccaatgcatt gggcccggta cccagctttt gttcccttta 5581 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg 5641 ttatccgctc acaattccac acaacatacg agccgggagc ataaagtgta aagcctgggg 5701 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc 5761 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt 5821 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct 5881 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga 5941 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc 6001 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg 6061 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg 6121 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt 6181 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt 6241 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg 6301 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact 6361 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt 6421 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct 6481 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac 6541 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc 6601 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg 6661 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta 6721 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca 6781 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc 6841 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc 6901 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc 6961 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat 7021 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt 7081 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc 7141 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag 7201 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt 7261 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac 7321 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg 7381 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat 7441 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc 7501 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc 7561 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa 7621 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg 7681 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg 7741 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg 7801 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct 7861 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt 7921 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat 7981 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca 8041 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac 8101 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta 8161 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg 8221 tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgc //