质粒名称: | BB3cH_pGAP_23*_pLAT1_Cas9 |
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出品公司: | Addgene |
目录编号: | 104907 |
存储实验室: | Brigitte Gasser |
载体骨架基本信息 |
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载体名称: | Episomal ARS/CEN Backbone |
载体出品公司: | Clontech |
空载体大小: | 4038 bp |
完整质粒大小: | 9323 bp |
载体类型: | Yeast Expression, CRISPR |
筛选标记: | Hygromycin |
载体抗性: | Hygromycin |
生长温度: | 37 ℃ |
宿主菌: | DH5a |
拷贝数: | 高拷贝 |
插入基因信息 |
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插入基因名称: | HH - FS23 linker - HDV |
克隆方法: | 限制性内切酶 |
5’克隆位点: | BbsI (not destroyed) |
3’克隆位点: | BbsI (not destroyed) |
5’测序引物 | GAP fw |
插入基因名称: | human codon-optimized hcas9 sequence fused to a nuclear localization signal (NLS) / hcas9 |
物种来源: | H. sapiens (human); Streptococcus pyogenes, Pichia pastoris |
插入大小: | 4140bp |
文献引用方法 |
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材料和方法部分: | BB3cH_pGAP_23*_pLAT1_Cas9 was a gift from Brigitte Gasser (Addgene plasmid # 104907 ; http://n2t.net/addgene:104907 ; RRID:Addgene_104907) |
参考文献部分: | CRISPR/Cas9-Mediated Homology-Directed Genome Editing in Pichia pastoris. Gassler T, Heistinger L, Mattanovich D, Gasser B, Prielhofer R. Methods Mol Biol. 2019;1923:211-225. doi: 10.1007/978-1-4939-9024-5_9. 10.1007/978-1-4939-9024-5_9 PubMed 30737742 |
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BB3cH_pGAP_23*_pLAT1_Cas9载体质粒基本信息
BB3cH_pGAP_23*_pLAT1_Cas9质粒图谱载体图谱和BB3cH_pGAP_23*_pLAT1_Cas9载体序列质粒序列多克隆位点信息
BB3cH_pGAP_23*_pLAT1_Cas9质粒载体简介
BB3cH_pGAP_23*_pLAT1_Cas9质粒序列载体序列
LOCUS BB3cH_pGAP_23*_pLAT1_Cas9 9323 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION hCas9 under control of LAT1 for direct cloning of HH-sgRNA-HDV PCR products for episomal expression in P. pastoris and selection on Hyg. ACCESSION . VERSION . KEYWORDS BB3cH_pGAP_23*_pLAT1_Cas9 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9323) AUTHORS Gassler T, Heistinger L, Mattanovich D, Gasser B, Prielhofer R. TITLE CRISPR/Cas9-mediated homology directed genome editing in Pichia pastoris JOURNAL (2018) In Press REFERENCE 2 (bases 1 to 9323) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..9323 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin complement(62..650) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(142..161) /label=pBR322ori-F /note="pBR322 origin, forward primer" promoter 737..1213 /label=GAP promoter /note="promoter for Pichia pastoris glyceraldehyde-3-phosphate dehydrogenase" CDS 2757..6860 /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 6873..6893 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" terminator 6965..7212 /gene="S. cerevisiae CYC1" /label=CYC1 terminator /note="transcription terminator for CYC1" primer_bind complement(6981..6999) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" gene 7232..8807 /label=hphMX6 /note="yeast selectable marker conferring hygromycin resistance" promoter 7232..7575 /label=TEF promoter /note="Ashbya gossypii TEF promoter" CDS 7576..8604 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA KE" terminator 8610..8807 /label=TEF terminator /note="Ashbya gossypii TEF terminator" misc_feature 8819..9322 /label=CEN/ARS /note="S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence" ORIGIN 1 cgctgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 61 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 121 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 181 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 241 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 301 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 361 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 421 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 481 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 541 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 601 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 661 tgatcttttc tacggggtgg agcgcaaaaa accccgcccc tgacagggcg gggttttttc 721 gcgatcggag gatccttttt tgtagaaatg tcttggtgtc ctcgtccaat caggtagcca 781 tctctgaaat atctggctcc gttgcaactc cgaacgacct gctggcaacg taaaattctc 841 cggggtaaaa cttaaatgtg gagtaatgga accagaaacg tctcttccct tctctctcct 901 tccaccgccc gttaccgtcc ctaggaaatt ttactctgct ggagagcttc ttctacggcc 961 cccttgcagc aatgctcttc ccagcattac gttgcgggta aaacggaggt cgtgtacccg 1021 acctagcagc ccagggatgg aaaagtcccg gccgtcgctg gcaataatag cgggcggacg 1081 catgtcatga gattattgga aaccaccaga atcgaatata aaaggcgaac acctttccca 1141 attttggttt ctcctgaccc aaagacttta aatttaattt atttgtccct atttcaatca 1201 attgaacaac tatcaaaaca ccatggcgtc ttcggatccc tcgaggaaga cgcgctttta 1261 gtgtacatct gataatatag tactaccacg tatgataatg tagagaatag gcttccttgt 1321 cgagtgtgtt tgcagttttc ttgagtttca aggtttaaat gctggtatat tagttcatcg 1381 aaggtttcag ccaatagcac cttaaatcaa tcaaactaat tcgactctta cgaaagagcc 1441 tactgtgttt agtatcgaag tcgtttacct ttcatgttga atagcttcct ctctgaccct 1501 aacatttcaa gatcctccta aagttacccg gattgtgaaa ttctaatgat ccacctgccc 1561 aatgcatttt ttctttattc agtttacctt ttttacctaa tatacgagct tgttaaagta 1621 agtggcactg caatactagg cttattgttg atattatgat gaatcgtttt cacaaacttg 1681 atttcctgtg aactcaccat gtactaagga aaaaaacatg catcaccatc tgaatatttg 1741 accgctccgg ggagggaaga atatgaacca cgattgaaag actttgggtt tgccgattct 1801 aaaacactta ctgaacagaa aagaagtgaa ttattagaga gtttatgcaa cggagatcat 1861 gagcttagaa agaacgtagg ctgggcaaca acatccattt ctgcctgcga tatttccagt 1921 gcaatgctaa agccacataa tataggcaat tacaacttga atgagcaggc tcatgatgtc 1981 accatggaac tgatagataa agtcatccaa aaaggtgtca atttacggca cgtctacgta 2041 gatactgtcg gacctccagc atcataccaa aagaaactgc aacaaaggtt tcccgcactg 2101 ctgatcaccg tatctaaaaa agcagatgcc atctatccat gtgtttcaag tgcttcagtt 2161 tgcgccaagg tcactagaga cgtctgtttg agaacctcaa atggggagga gaacctatta 2221 gattgtgggt ctggatatcc ctcggaccca aatactaaga aatggctagc ccagtctata 2281 catccattgt ttggctggag tcgtgatgtc aggttttcgt ggcagacggc caaagattgt 2341 cttgagagac acaactcaat caaagttgtg tttggcgaag aggagcctac cccggttaat 2401 ttcggtgatc tggggttgag caaacagaag cccgacttta aggatttaga aatcagtacc 2461 agttggtggg gacattaatt ggtggaggag agccaatatt ctcaactgtt cgtaggacca 2521 taacccccca ctcgcaccag aaatactcat tggcgctgac aactagtttg agccgcttca 2581 attgcggcaa gtccccagtt ttgtttggta ctcccctctc gcctcgcgtg tttttgttcc 2641 tccgaattgt gcatcaccct gtcacatccg tacaccatcg gaacactctg ctcctcctcc 2701 tttctcctct taattgaacg tatatctaat taccctaata acaagtatct tcaaccatgg 2761 acaagaagta ctccattggg ctcgatatcg gcacaaacag cgtcggttgg gccgtcatta 2821 cggacgagta caaggtgccg agcaaaaaat tcaaagttct gggcaatacc gatcgccaca 2881 gcataaagaa gaacctcatt ggcgccctcc tgttcgactc cggggagacg gccgaagcca 2941 cgcggctcaa aagaacagca cggcgcagat atacccgcag aaagaatcgg atctgctacc 3001 tgcaggagat ctttagtaat gagatggcta aggtggatga ctctttcttc cataggctgg 3061 aggagtcctt tttggtggag gaggataaaa agcacgagcg ccacccaatc tttggcaata 3121 tcgtggacga ggtggcgtac catgaaaagt acccaaccat atatcatctg aggaagaagc 3181 ttgtagacag tactgataag gctgacttgc ggttgatcta tctcgcgctg gcgcatatga 3241 tcaaatttcg gggacacttc ctcatcgagg gggacctgaa cccagacaac agcgatgtcg 3301 acaaactctt tatccaactg gttcagactt acaatcagct tttcgaagag aacccgatca 3361 acgcatccgg agttgacgcc aaagcaatcc tgagcgctag gctgtccaaa tcccggcggc 3421 tcgaaaacct catcgcacag ctccctgggg agaagaagaa cggcctgttt ggtaatctta 3481 tcgccctgtc actcgggctg acccccaact ttaaatctaa cttcgacctg gccgaagatg 3541 ccaagcttca actgagcaaa gacacctacg atgatgatct cgacaatctg ctggcccaga 3601 tcggcgacca gtacgcagac ctttttttgg cggcaaagaa cctgtcagac gccattctgc 3661 tgagtgatat tctgcgagtg aacacggaga tcaccaaagc tccgctgagc gctagtatga 3721 tcaagcgcta tgatgagcac caccaagact tgactttgct gaaggccctt gtcagacagc 3781 aactgcctga gaagtacaag gaaattttct tcgatcagtc taaaaatggc tacgccggat 3841 acattgacgg cggagcaagc caggaggaat tttacaaatt tattaagccc atcttggaaa 3901 aaatggacgg caccgaggag ctgctggtaa agcttaacag agaagatctg ttgcgcaaac 3961 agcgcacttt cgacaatgga agcatccccc accagattca cctgggcgaa ctgcacgcta 4021 tcctcaggcg gcaagaggat ttctacccct ttttgaaaga taacagggaa aagattgaga 4081 aaatcctcac atttcggata ccctactatg taggccccct cgcccgggga aattccagat 4141 tcgcgtggat gactcgcaaa tcagaagaga ctatcactcc ctggaacttc gaggaagtcg 4201 tggataaggg ggcctctgcc cagtccttca tcgaaaggat gactaacttt gataaaaatc 4261 tgcctaacga aaaggtgctt cctaaacact ctctgctgta cgagtacttc acagtttata 4321 acgagctcac caaggtcaaa tacgtcacag aagggatgag aaagccagca ttcctgtctg 4381 gagagcagaa gaaagctatc gtggacctcc tcttcaagac gaaccggaaa gttaccgtga 4441 aacagctcaa agaggactat ttcaaaaaga ttgaatgttt cgactctgtt gaaatcagcg 4501 gagtggagga tcgcttcaac gcatccctgg gaacgtatca cgatctcctg aaaatcatta 4561 aagacaagga cttcctggac aatgaggaga acgaggacat tcttgaggac attgtcctca 4621 cccttacgtt gtttgaagat agggagatga ttgaagaacg cttgaaaact tacgctcatc 4681 tcttcgacga caaagtcatg aaacagctca agaggcgccg atatacagga tgggggcggc 4741 tgtcaagaaa actgatcaat gggatccgag acaagcagag tggaaagaca atcctggatt 4801 ttcttaagtc cgatggattt gccaaccgga acttcatgca gttgatccat gatgactctc 4861 tcacctttaa ggaggacatc cagaaagcac aagtttctgg ccagggggac agccttcacg 4921 agcacatcgc taatcttgca ggtagcccag ctatcaaaaa gggaatactg cagaccgtta 4981 aggtcgtgga tgaactcgtc aaagtaatgg gaaggcataa gcccgagaat atcgttatcg 5041 agatggcccg agagaaccaa actacccaga agggacagaa gaacagtagg gaaaggatga 5101 agaggattga agagggtata aaagaactgg ggtcccaaat ccttaaggaa cacccagttg 5161 aaaacaccca gcttcagaat gagaagctct acctgtacta cctgcagaac ggcagggaca 5221 tgtacgtgga tcaggaactg gacatcaatc ggctctccga ctacgacgtg gatcatatcg 5281 tgccccagtc ttttctcaaa gatgattcta ttgataataa agtgttgaca agatccgata 5341 aaaatagagg gaagagtgat aacgtcccct cagaagaagt tgtcaagaaa atgaaaaatt 5401 attggcggca gctgctgaac gccaaactga tcacacaacg gaagttcgat aatctgacta 5461 aggctgaacg aggtggcctg tctgagttgg ataaagccgg cttcatcaaa aggcagcttg 5521 ttgagacacg ccagatcacc aagcacgtgg cccaaattct cgattcacgc atgaacacca 5581 agtacgatga aaatgacaaa ctgattcgag aggtgaaagt tattactctg aagtctaagc 5641 tcgtctcaga tttcagaaag gactttcagt tttataaggt gagagagatc aacaattacc 5701 accatgcgca tgatgcctac ctgaatgcag tggtaggcac tgcacttatc aaaaaatatc 5761 ccaagcttga atctgaattt gtttacggag actataaagt gtacgatgtt aggaaaatga 5821 tcgcaaagtc tgagcaggaa ataggcaagg ccaccgctaa gtacttcttt tacagcaata 5881 ttatgaattt tttcaagacc gagattacac tggccaatgg agagattcgg aagcgaccac 5941 ttatcgaaac aaacggagaa acaggagaaa tcgtgtggga caagggtagg gatttcgcga 6001 cagtccggaa ggtcctgtcc atgccgcagg tgaacatcgt taaaaagacc gaagtacaga 6061 ccggaggctt ctccaaggaa agtatcctcc cgaaaaggaa cagcgacaag ctgatcgcac 6121 gcaaaaaaga ttgggacccc aagaaatacg gcggattcga ttctcctaca gtcgcttaca 6181 gtgtactggt tgtggccaaa gtggagaaag ggaagtctaa aaaactcaaa agcgtcaagg 6241 aactgctggg catcacaatc atggagcgat caagcttcga aaaaaacccc atcgactttc 6301 tcgaggcgaa aggatataaa gaggtcaaaa aagacctcat cattaagctt cccaagtact 6361 ctctctttga gcttgaaaac ggccggaaac gaatgctcgc tagtgcgggc gagctgcaga 6421 aaggtaacga gctggcactg ccctctaaat acgttaattt cttgtatctg gccagccact 6481 atgaaaagct caaagggtcc cccgaagata atgagcagaa gcagctgttc gtggaacaac 6541 acaaacacta ccttgatgag atcatcgagc aaataagcga attctccaaa agagtgatcc 6601 tcgccgacgc taacctcgat aaggtgcttt ctgcttacaa taagcacagg gataagccca 6661 tcagggagca ggcagaaaac attatccact tgtttactct gaccaacttg ggcgcgcctg 6721 cagccttcaa gtacttcgac accaccatag acagaaagcg gtacacctct acaaaggagg 6781 tcctggacgc cacactgatt catcagtcaa ttacggggct ctatgaaaca agaatcgacc 6841 tctctcagct cggtggagac agcagggctg accccaagaa gaagaggaag gtgtgagctt 6901 cgtccgacgg cggcccgacg ggtccgaggc ctcggagatc cgtccccctt ttcctttgtc 6961 gatatcatgt aattagttat gtcacgctta cattcacgcc ctccccccac atccgctcta 7021 accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt ttttatagtt 7081 atgttagtat taagaacgtt atttatattt caaatttttc ttttttttct gtacagacgc 7141 gtgtacgcat gtaacattat actgaaaacc ttgcttgaga aggttttggg acgctcgaag 7201 gctttaattt gcaagctggc gctaattcat ggacatggag gcccagaata ccctccttga 7261 cagtcttgac gtgcgcagct caggggcatg atgtgactgt cgcccgtaca tttagcccat 7321 acatccccat gtataatcat ttgcatccat acattttgat ggccgcacgg cgcgaagcaa 7381 aaattacggc tcctcgctgc agacgtgcga gcagggaaac gctcccctca cagacgcgtt 7441 gaattgtccc cacgccgcgc ccctgtagag aaatataaaa ggttaggatt tgccactgag 7501 gttcttcttt catatacttc cttttaaaat cttgctagga tacagttctc acatcacatc 7561 cgaacataaa caaccatggg taaaaagcct gaactcaccg cgacgtctgt cgagaagttt 7621 ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct 7681 cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc 7741 gatggtttct acaaagatcg ttatgtttat cggcactttg catcggccgc gctcccgatt 7801 ccggaagtgc ttgacattgg ggaattcagc gagagcctga cctattgcat ctcccgccgt 7861 gcacagggtg tcacgttgca agacctccct gaaaccgaac tgcccgctgt tctgcagccg 7921 gtcgcggagg ccatggatgc gatcgctgcg gccgatctta gccagacgag cgggttcggc 7981 ccattcggac cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt 8041 gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc 8101 gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc 8161 gtgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc 8221 attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc 8281 tggaggccgt ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg 8341 gagcttgcag gatcgccgcg gctccgggcg tatatgctcc gcattggtct tgaccaactc 8401 tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac 8461 gcaatcgtcc gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg 8521 gccgtctgga ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc 8581 actcgtccga gggcaaagga ataatcagta ctgacaataa aaagattctt gttttcaaga 8641 acttgtcatt tgtatagttt ttttatattg tagttgttct attttaatca aatgttagcg 8701 tgatttatat tttttttcgc ctcgacatca tctgcccaga tgcgaagtta agtgcgcaga 8761 aagtaatatc atgcgtcaat cgtatgtgaa tgctggtcgc tatactggct tccttttcat 8821 cacgtgctat aaaaataatt ataatttaaa ttttttaata taaatatata aattaaaaat 8881 agaaagtaaa aaaagaaatt aaagaaaaaa tagtttttgt tttccgaaga tgtaaaagac 8941 tctaggggga tcgccaacaa atactacctt ttatcttgct cttcctgctc tcaggtatta 9001 atgccgaatt gtttcatctt gtctgtgtag aacaccacac acgaaaatcc tgtgatttta 9061 cattttactt atcgttaatc gaatgtatat ctatttaatc tgcttttctt gtctaataaa 9121 tatatatgta aagtacgctt tttgttgaaa ttttttaaac ctttgtttat ttttttttct 9181 tcattccgta actcttctac cttctttatt tactttctaa aatccaaata caaaacataa 9241 aaataaataa acacagagta aattcccaaa ttattccatc attaaaagat acgaggcgcg 9301 tgtaagttac aggcaagcga tcg //