质粒名称: | p414-TEF1p-Cas9-CYC1t |
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出品公司: | 爱迪基因 |
目录编号: | 43802 |
存储实验室: | George Church |
载体骨架基本信息 |
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载体名称: | p414 |
载体抗性: | 氨苄西林 |
空载体大小: | 5385 bp |
完整质粒大小: | 9525 bp |
载体类型: | 酵母表达,crispr |
筛选标记: | 色氨酸 |
高拷贝/低拷贝: | 低拷贝 |
生长条件: | 30°C |
插入基因信息 |
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插入基因名称: | Human Optimized S. pyogenes Cas9 |
物种来源: | H.Sapiens(人类),S.cerevisiae(发芽酵母);S.化脓性 |
插入大小: | 4140 bp |
启动子 | TEF1 promoter |
5' 测序引物 | T3 promoter |
3' 测序引物 | T7 promoter |
文献引用方法 |
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材料和方法部分: | p414-TEF1p-Cas9-CYC1t was a gift from George Church (Addgene plasmid # 43802 ; http://n2t.net/addgene:43802 ; RRID:Addgene_43802) |
参考文献部分: | Genome engineering in Saccharomyces cerevisiae using CRISPR-Cas systems. Dicarlo JE, Norville JE, Mali P, Rios X, Aach J, Church GM.. Nucleic Acids Res 10.1093/nar/gkt135 PubMed 23460208 |
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p414-TEF1p-Cas9-CYC1t载体质粒基本信息
p414-TEF1p-Cas9-CYC1t质粒图谱载体图谱和p414-TEF1p-Cas9-CYC1t载体序列质粒序列多克隆位点信息
p414-TEF1p-Cas9-CYC1t质粒载体简介
p414-TEF1p-Cas9-CYC1t质粒序列载体序列
LOCUS p414-TEF1p-Cas9-CYC1t 9524 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS p414-TEF1p-Cas9-CYC1t SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9524) AUTHORS Dicarlo JE, Norville JE, Mali P, Rios X, Aach J, Church GM. TITLE Genome engineering in Saccharomyces cerevisiae using CRISPR-Cas systems. JOURNAL Nucleic Acids Res PUBMED 23460208 REFERENCE 2 (bases 1 to 9524) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9524 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(10..28) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" misc_feature 69..572 /label=CEN/ARS /note="S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence" promoter 609..713 /gene="bla" /label=AmpR promoter CDS 714..1574 /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 complement(932..951) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1745..2333 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2234..2253 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2487..2504 /label=L4440 /note="L4440 vector, forward primer" promoter 2657..2687 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2695..2711 /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)." primer_bind 2700..2722 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 2719..2735 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 2719..2735 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 2754..2774 /label=T3 /note="T3 promoter, forward primer" promoter 2756..2774 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 2792..3190 /gene="S. cerevisiae TEF1" /label=TEF1 promoter /note="promoter for EF-1-alpha" primer_bind 3194..3210 /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind 3194..3210 /label=pBluescriptSK /note="For pBluescript vector" CDS 3222..7325 /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 7338..7358 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" terminator 7374..7621 /gene="S. cerevisiae CYC1" /label=CYC1 terminator /note="transcription terminator for CYC1" primer_bind complement(7390..7408) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" primer_bind complement(7639..7658) /label=T7 /note="T7 promoter, forward primer" promoter complement(7640..7658) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(7668..7685) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(7668..7684) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7677..7699) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 7829..8284 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(7916..7935) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 8126..8147 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(8383..9057) /codon_start=1 /gene="S. cerevisiae TRP1" /product="phosphoribosylanthranilate isomerase, required for tryptophan biosynthesis" /label=TRP1 /note="yeast auxotrophic marker" /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA KK" promoter complement(9058..9338) /gene="S. cerevisiae TRP1" /label=TRP1 promoter primer_bind complement(9355..9374) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 9474..9496 /label=pGEX 3' /note="pGEX vectors, reverse primer" ORIGIN 1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt 61 cttagacgga tcgcttgcct gtaacttaca cgcgcctcgt atcttttaat gatggaataa 121 tttgggaatt tactctgtgt ttatttattt ttatgttttg tatttggatt ttagaaagta 181 aataaagaag gtagaagagt tacggaatga agaaaaaaaa ataaacaaag gtttaaaaaa 241 tttcaacaaa aagcgtactt tacatatata tttattagac aagaaaagca gattaaatag 301 atatacattc gattaacgat aagtaaaatg taaaatcaca ggattttcgt gtgtggtctt 361 ctacacagac aagatgaaac aattcggcat taatacctga gagcaggaag agcaagataa 421 aaggtagtat ttgttggcga tccccctaga gtcttttaca tcttcggaaa acaaaaacta 481 ttttttcttt aatttctttt tttactttct atttttaatt tatatattta tattaaaaaa 541 tttaaattat aattattttt atagcacgtg atgaaaagga cccaggtggc acttttcggg 601 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc 661 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta 721 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg 781 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg 841 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac 901 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg 961 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt 1021 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg 1081 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac 1141 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt 1201 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag 1261 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc 1321 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc 1381 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta 1441 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg 1501 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga 1561 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac 1621 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa 1681 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat 1741 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc 1801 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg 1861 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc 1921 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg 1981 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg 2041 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa 2101 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg 2161 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga 2221 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct 2281 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca 2341 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc 2401 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg 2461 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc 2521 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca 2581 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tacctcactc 2641 attaggcacc ccaggcttta cactttatgc ttccggctcc tatgttgtgt ggaattgtga 2701 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta 2761 accctcacta aagggaacaa aagctggagc tcatagcttc aaaatgtttc tactcctttt 2821 ttactcttcc agattttctc ggactccgcg catcgccgta ccacttcaaa acacccaagc 2881 acagcatact aaatttcccc tctttcttcc tctagggtgt cgttaattac ccgtactaaa 2941 ggtttggaaa agaaaaaaga gaccgcctcg tttctttttc ttcgtcgaaa aaggcaataa 3001 aaatttttat cacgtttctt tttcttgaaa attttttttt tgattttttt ctctttcgat 3061 gacctcccat tgatatttaa gttaataaac ggtcttcaat ttctcaagtt tcagtttcat 3121 ttttcttgtt ctattacaac tttttttact tcttgctcat tagaaagaaa gcatagcaat 3181 ctaatctaag ttttctagaa ctagtggatc ccccgggaaa aatggacaag aagtactcca 3241 ttgggctcga tatcggcaca aacagcgtcg gttgggccgt cattacggac gagtacaagg 3301 tgccgagcaa aaaattcaaa gttctgggca ataccgatcg ccacagcata aagaagaacc 3361 tcattggcgc cctcctgttc gactccgggg agacggccga agccacgcgg ctcaaaagaa 3421 cagcacggcg cagatatacc cgcagaaaga atcggatctg ctacctgcag gagatcttta 3481 gtaatgagat ggctaaggtg gatgactctt tcttccatag gctggaggag tcctttttgg 3541 tggaggagga taaaaagcac gagcgccacc caatctttgg caatatcgtg gacgaggtgg 3601 cgtaccatga aaagtaccca accatatatc atctgaggaa gaagcttgta gacagtactg 3661 ataaggctga cttgcggttg atctatctcg cgctggcgca tatgatcaaa tttcggggac 3721 acttcctcat cgagggggac ctgaacccag acaacagcga tgtcgacaaa ctctttatcc 3781 aactggttca gacttacaat cagcttttcg aagagaaccc gatcaacgca tccggagttg 3841 acgccaaagc aatcctgagc gctaggctgt ccaaatcccg gcggctcgaa aacctcatcg 3901 cacagctccc tggggagaag aagaacggcc tgtttggtaa tcttatcgcc ctgtcactcg 3961 ggctgacccc caactttaaa tctaacttcg acctggccga agatgccaag cttcaactga 4021 gcaaagacac ctacgatgat gatctcgaca atctgctggc ccagatcggc gaccagtacg 4081 cagacctttt tttggcggca aagaacctgt cagacgccat tctgctgagt gatattctgc 4141 gagtgaacac ggagatcacc aaagctccgc tgagcgctag tatgatcaag cgctatgatg 4201 agcaccacca agacttgact ttgctgaagg cccttgtcag acagcaactg cctgagaagt 4261 acaaggaaat tttcttcgat cagtctaaaa atggctacgc cggatacatt gacggcggag 4321 caagccagga ggaattttac aaatttatta agcccatctt ggaaaaaatg gacggcaccg 4381 aggagctgct ggtaaagctt aacagagaag atctgttgcg caaacagcgc actttcgaca 4441 atggaagcat cccccaccag attcacctgg gcgaactgca cgctatcctc aggcggcaag 4501 aggatttcta cccctttttg aaagataaca gggaaaagat tgagaaaatc ctcacatttc 4561 ggatacccta ctatgtaggc cccctcgccc ggggaaattc cagattcgcg tggatgactc 4621 gcaaatcaga agagaccatc actccctgga acttcgagga agtcgtggat aagggggcct 4681 ctgcccagtc cttcatcgaa aggatgacta actttgataa aaatctgcct aacgaaaagg 4741 tgcttcctaa acactctctg ctgtacgagt acttcacagt ttataacgag ctcaccaagg 4801 tcaaatacgt cacagaaggg atgagaaagc cagcattcct gtctggagag cagaagaaag 4861 ctatcgtgga cctcctcttc aagacgaacc ggaaagttac cgtgaaacag ctcaaagaag 4921 actatttcaa aaagattgaa tgtttcgact ctgttgaaat cagcggagtg gaggatcgct 4981 tcaacgcatc cctgggaacg tatcacgatc tcctgaaaat cattaaagac aaggacttcc 5041 tggacaatga ggagaacgag gacattcttg aggacattgt cctcaccctt acgttgtttg 5101 aagataggga gatgattgaa gaacgcttga aaacttacgc tcatctcttc gacgacaaag 5161 tcatgaaaca gctcaagagg cgccgatata caggatgggg gcggctgtca agaaaactga 5221 tcaatgggat ccgagacaag cagagtggaa agacaatcct ggattttctt aagtccgatg 5281 gatttgccaa ccggaacttc atgcagttga tccatgatga ctctctcacc tttaaggagg 5341 acatccagaa agcacaagtt tctggccagg gggacagtct tcacgagcac atcgctaatc 5401 ttgcaggtag cccagctatc aaaaagggaa tactgcagac cgttaaggtc gtggatgaac 5461 tcgtcaaagt aatgggaagg cataagcccg agaatatcgt tatcgagatg gcccgagaga 5521 accaaactac ccagaaggga cagaagaaca gtagggaaag gatgaagagg attgaagagg 5581 gtataaaaga actggggtcc caaatcctta aggaacaccc agttgaaaac acccagcttc 5641 agaatgagaa gctctacctg tactacctgc agaacggcag ggacatgtac gtggatcagg 5701 aactggacat caatcggctc tccgactacg acgtggatca tatcgtgccc cagtcttttc 5761 tcaaagatga ttctattgat aataaagtgt tgacaagatc cgataaaaat agagggaaga 5821 gtgataacgt cccctcagaa gaagttgtca agaaaatgaa aaattattgg cggcagctgc 5881 tgaacgccaa actgatcaca caacggaagt tcgataatct gactaaggct gaacgaggtg 5941 gcctgtctga gttggataaa gccggcttca tcaaaaggca gcttgttgag acacgccaga 6001 tcaccaagca cgtggcccaa attctcgatt cacgcatgaa caccaagtac gatgaaaatg 6061 acaaactgat tcgagaggtg aaagttatta ctctgaagtc taagctggtc tcagatttca 6121 gaaaggactt tcagttttat aaggtgagag agatcaacaa ttaccaccat gcgcatgatg 6181 cctacctgaa tgcagtggta ggcactgcac ttatcaaaaa atatcccaag cttgaatctg 6241 aatttgttta cggagactat aaagtgtacg atgttaggaa aatgatcgca aagtctgagc 6301 aggaaatagg caaggccacc gctaagtact tcttttacag caatattatg aattttttca 6361 agaccgagat tacactggcc aatggagaga ttcggaagcg accacttatc gaaacaaacg 6421 gagaaacagg agaaatcgtg tgggacaagg gtagggattt cgcgacagtc cggaaggtcc 6481 tgtccatgcc gcaggtgaac atcgttaaaa agaccgaagt acagaccgga ggcttctcca 6541 aggaaagtat cctcccgaaa aggaacagcg acaagctgat cgcacgcaaa aaagattggg 6601 accccaagaa atacggcgga ttcgattctc ctacagtcgc ttacagtgta ctggttgtgg 6661 ccaaagtgga gaaagggaag tctaaaaaac tcaaaagcgt caaggaactg ctgggcatca 6721 caatcatgga gcgatcaagc ttcgaaaaaa accccatcga ctttctcgag gcgaaaggat 6781 ataaagaggt caaaaaagac ctcatcatta agcttcccaa gtactctctc tttgagcttg 6841 aaaacggccg gaaacgaatg ctcgctagtg cgggcgagct gcagaaaggt aacgagctgg 6901 cactgccctc taaatacgtt aatttcttgt atctggccag ccactatgaa aagctcaaag 6961 ggtctcccga agataatgag cagaagcagc tgttcgtgga acaacacaaa cactaccttg 7021 atgagatcat cgagcaaata agcgaattct ccaaaagagt gatcctcgcc gacgctaacc 7081 tcgataaggt gctttctgct tacaataagc acagggataa gcccatcagg gagcaggcag 7141 aaaacattat ccacttgttt actctgacca acttgggcgc gcctgcagcc ttcaagtact 7201 tcgacaccac catagacaga aagcggtaca cctctacaaa ggaggtcctg gacgccacac 7261 tgattcatca gtcaattacg gggctctatg aaacaagaat cgacctctct cagctcggtg 7321 gagacagcag ggctgacccc aagaagaaga ggaaggtgtg atctcttctc gagtcatgta 7381 attagttatg tcacgcttac attcacgccc tccccccaca tccgctctaa ccgaaaagga 7441 aggagttaga caacctgaag tctaggtccc tatttatttt tttatagtta tgttagtatt 7501 aagaacgtta tttatatttc aaatttttct tttttttctg tacagacgcg tgtacgcatg 7561 taacattata ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg ctttaatttg 7621 cggccggtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt 7681 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat 7741 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag 7801 ttgcgcagcc tgaatggcga atggcgcgac gcgccctgta gcggcgcatt aagcgcggcg 7861 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct 7921 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat 7981 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt 8041 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg 8101 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac 8161 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta 8221 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca 8281 atttcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcataggca 8341 agtgcacaaa caatacttaa ataaatacta ctcagtaata acctatttct tagcattttt 8401 gacgaaattt gctattttgt tagagtcttt tacaccattt gtctccacac ctccgcttac 8461 atcaacacca ataacgccat ttaatctaag cgcatcacca acattttctg gcgtcagtcc 8521 accagctaac ataaaatgta agctttcggg gctctcttgc cttccaaccc agtcagaaat 8581 cgagttccaa tccaaaagtt cacctgtccc acctgcttct gaatcaaaca agggaataaa 8641 cgaatgaggt ttctgtgaag ctgcactgag tagtatgttg cagtcttttg gaaatacgag 8701 tcttttaata actggcaaac cgaggaactc ttggtattct tgccacgact catctccatg 8761 cagttggacg atatcaatgc cgtaatcatt gaccagagcc aaaacatcct ccttaggttg 8821 attacgaaac acgccaacca agtatttcgg agtgcctgaa ctatttttat atgcttttac 8881 aagacttgaa attttccttg caataaccgg gtcaattgtt ctctttctat tgggcacaca 8941 tataataccc agcaagtcag catcggaatc tagagcacat tctgcggcct ctgtgctctg 9001 caagccgcaa actttcacca atggaccaga actacctgtg aaattaataa cagacatact 9061 ccaagctgcc tttgtgtgct taatcacgta tactcacgtg ctcaatagtc accaatgccc 9121 tccctcttgg ccctctcctt ttcttttttc gaccgaatta attcttaatc ggcaaaaaaa 9181 gaaaagctcc ggatcaagat tgtacgtaag gtgacaagct atttttcaat aaagaatatc 9241 ttccactact gccatctggc gtcataactg caaagtacac atatattacg atgctgtcta 9301 ttaaatgctt cctatattat atatatagta atgtcgttta tggtgcactc tcagtacaat 9361 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc 9421 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag 9481 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcga //