质粒名称: | pdCas9-PBE |
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出品公司: | 爱迪基因 |
目录编号: | 98161 |
存储实验室: | Caixia Gao |
载体骨架基本信息 |
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载体名称: | pJIT163-Ubi-GFP |
载体出品公司: | -- |
载体抗性: | 氨苄青霉素 |
空载体大小: | -- |
完整质粒大小: | -- |
载体修饰: | -- |
载体类型: | 植物表达 |
筛选标记: | -- |
高拷贝/低拷贝: | -- |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | NLS-APOBEC1-XTEN-dCas9-UGI-NLS |
物种来源: | -- |
插入大小: | -- |
基因突变: | -- |
GenBank编号 | -- |
启动子 | Ubi |
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: | Bsp1407I (未破坏) |
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: | pdCas9-PBE was a gift from Caixia Gao (Addgene plasmid # 98161 ; http://n2t.net/addgene:98161 ; RRID:Addgene_98161) |
参考文献部分: | Precise base editing in rice, wheat and maize with a Cas9-cytidine deaminase fusion. Zong Y, Wang Y, Li C, Zhang R, Chen K, Ran Y, Qiu JL, Wang D, Gao C. Nat Biotechnol. 2017 May;35(5):438-440. doi: 10.1038/nbt.3811. Epub 2017 Feb 27. 10.1038/nbt.3811 PubMed 28244994 |
买家导航
pdCas9PBE载体基本信息
pdCas9PBE载体图谱质粒图谱和多克隆位点信息
pdCas9PBE载体简介
pdCas9PBE载体序列
LOCUS pdCas9-PBE 10201 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION Expresses CRISPR base editor in protoplasts. ACCESSION . VERSION . KEYWORDS pdCas9-PBE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10201) AUTHORS Zong Y, Wang Y, Li C, Zhang R, Chen K, Ran Y, Qiu JL, Wang D, Gao C TITLE Precise base editing in rice, wheat and maize with a Cas9-cytidine deaminase fusion. JOURNAL Nat Biotechnol. 2017 May;35(5):438-440. doi: 10.1038/nbt.3811. Epub 2017 Feb 27. PUBMED 28244994 REFERENCE 2 (bases 1 to 10201) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10201 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 2020..2040 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 2041..2724 /codon_start=1 /product="cytidine deaminase (C to U editing enzyme) from rat" /label=APOBEC-1 /note="can use ssDNA as a substrate (Komor et al., 2016)" /translation="SSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGG RHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLS RYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNE AHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHIL WATGLK" CDS 2776..6876 /codon_start=1 /product="catalytically dead mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=dCas9 /note="RNA-guided DNA-binding protein that lacks endonuclease activity due to the D10A mutation in the RuvC catalytic domain and the H840A mutation in the HNH catalytic domain" /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD ATLIHQSITGLYETRIDLSQLGGD" CDS 6877..6924 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" CDS 6946..7194 /codon_start=1 /product="uracil-DNA glycosylase inhibitor from a Bacillus subtilis bacteriophage (Mol et al., 1995)" /label=UGI /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML" CDS 7207..7227 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" polyA_signal 7301..7477 /label=CaMV poly(A) signal /note="cauliflower mosaic virus polyadenylation signal" primer_bind complement(8031..8047) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(8031..8047) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(8044..8066) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 8055..8071 /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(8079..8109) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8124..8145 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(8262..8279) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(8433..9021) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(8513..8532) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(9192..10052) /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 9815..9834 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(10053..10157) /gene="bla" /label=AmpR promoter ORIGIN 1 gagctcggta cctgacccgg tcgtgcccct ctctagagat aatgagcatt gcatgtctaa 61 gttataaaaa attaccacat attttttttg tcacacttgt ttgaagtgca gtttatctat 121 ctttatacat atatttaaac tttactctac gaataatata atctatagta ctacaataat 181 atcagtgttt tagagaatca tataaatgaa cagttagaca tggtctaaag gacaattgag 241 tattttgaca acaggactct acagttttat ctttttagtg tgcatgtgtt ctcctttttt 301 tttgcaaata gcttcaccta tataatactt catccatttt attagtacat ccatttaggg 361 tttagggtta atggttttta tagactaatt tttttagtac atctatttta ttctatttta 421 gcctctaaat taagaaaact aaaactctat tttagttttt ttatttaata atttagatat 481 aaaatagaat aaaataaagt gactaaaaat taaacaaata ccctttaaga aattaaaaaa 541 actaaggaaa catttttctt gtttcgagta gataatgcca gcctgttaaa cgccgtcgac 601 gagtctaacg gacaccaacc agcgaaccag cagcgtcgcg tcgggccaag cgaagcagac 661 ggcacggcat ctctgtcgct gcctctggac ccctctcgat cgagagttcc gctccaccgt 721 tggacttgct ccgctgtcgg catccagaaa ttgcgtggcg gagcggcaga cgtgagccgg 781 cacggcaggc ggcctcctcc tcctctcacg gcaccggcag ctacggggga ttcctttccc 841 accgctcctt cgctttccct tcctcgcccg ccgtaataaa tagacacccc ctccacaccc 901 tctttcccca acctcgtgtt gttcggagcg cacacacaca caaccagatc tcccccaaat 961 ccacccgtcg gcacctccgc ttcaaggtac gccgctcgtc ctcccccccc cccctctcta 1021 ccttctctag atcggcgttc cggtccatgg ttagggcccg gtagttctac ttctgttcat 1081 gtttgtgtta gatccgtgtt tgtgttagat ccgtgctgct agcgttcgta cacggatgcg 1141 acctgtacgt cagacacgtt ctgattgcta acttgccagt gtttctcttt ggggaatcct 1201 gggatggctc tagccgttcc gcagacggga tcgatttcat gatttttttt gtttcgttgc 1261 atagggtttg gtttgccctt ttcctttatt tcaatatatg ccgtgcactt gtttgtcggg 1321 tcatcttttc atgctttttt ttgtcttggt tgtgatgatg tggtctggtt gggcggtcgt 1381 tctagatcgg agtagaatta attctgtttc aaactacctg gtggatttat taattttgga 1441 tctgtatgtg tgtgccatac atattcatag ttacgaattg aagatgatgg atggaaatat 1501 cgatctagga taggtataca tgttgatgcg ggttttactg atgcatatac agagatgctt 1561 tttgttcgct tggttgtgat gatgtggtgt ggttgggcgg tcgttcattc gttctagatc 1621 ggagtagaat actgtttcaa actacctggt gtatttatta attttggaac tgtatgtgtg 1681 tgtcatacat cttcatagtt acgagtttaa gatggatgga aatatcgatc taggataggt 1741 atacatgttg atgtgggttt tactgatgca tatacatgat ggcatatgca gcatctattc 1801 atatgctcta accttgagta cctatctatt ataataaaca agtatgtttt ataattattt 1861 tgatcttgat atacttggat gatggcatat gcagcagcta tatgtggatt tttttagccc 1921 tgccttcata cgctatttat ttgcttggta ctgtttcttt tgtcgatgct caccctgttg 1981 tttggtgtta cttctgcaaa gcttgtcgac ggatccatgc caaagaagaa gaggaaggtt 2041 tcatcggaga ccggccctgt tgctgttgac cccaccctgc ggcggagaat cgagccacac 2101 gagttcgagg tgttcttcga cccaagggag ctccgcaagg agacgtgcct cctgtacgag 2161 atcaactggg gcggcaggca ctccatctgg aggcacacca gccaaaacac caacaagcac 2221 gtggaggtca acttcatcga gaagttcacc accgagaggt acttctgccc aaacacccgc 2281 tgctccatca cctggttcct gtcctggagc ccatgcggcg agtgctccag ggccatcacc 2341 gagttcctca gccgctaccc acacgtcacc ctgttcatct acatcgccag gctctaccac 2401 cacgccgacc caaggaacag gcagggcctc cgcgacctga tctccagcgg cgtgaccatc 2461 caaatcatga ccgagcagga gtccggctac tgctggagga acttcgtcaa ctactcccca 2521 agcaacgagg cccactggcc aaggtaccca cacctctggg tgcgcctcta cgtgctcgag 2581 ctgtactgca tcatcctcgg cctgccacca tgcctcaaca tcctgaggcg caagcaacca 2641 cagctgacct tcttcaccat cgccctccaa agctgccact accagaggct cccaccacac 2701 atcctgtggg ctaccggcct caagtccggc agcgagacgc caggcacctc cgagagcgct 2761 acgcctgaac ttaaggacaa gaagtactcg atcggcctcg ccatcgggac gaactcagtt 2821 ggctgggccg tgatcaccga cgagtacaag gtgccctcta agaagttcaa ggtcctgggg 2881 aacaccgacc gccattccat caagaagaac ctcatcggcg ctctcctgtt cgacagcggg 2941 gagaccgctg aggctacgag gctcaagaga accgctaggc gccggtacac gagaaggaag 3001 aacaggatct gctacctcca agagattttc tccaacgaga tggccaaggt tgacgattca 3061 ttcttccacc gcctggagga gtctttcctc gtggaggagg ataagaagca cgagcggcat 3121 cccatcttcg gcaacatcgt ggacgaggtt gcctaccacg agaagtaccc tacgatctac 3181 catctgcgga agaagctcgt ggactccacc gataaggcgg acctcagact gatctacctc 3241 gctctggccc acatgatcaa gttccgcggc catttcctga tcgaggggga tctcaaccca 3301 gacaacagcg atgttgacaa gctgttcatc caactcgtgc agacctacaa ccaactcttc 3361 gaggagaacc cgatcaacgc ctctggcgtg gacgcgaagg ctatcctgtc cgcgaggctc 3421 tcgaagtcca ggaggctgga gaacctgatc gctcagctcc caggcgagaa gaagaacggc 3481 ctgttcggga acctcatcgc tctcagcctg gggctcaccc cgaacttcaa gtcgaacttc 3541 gatctcgctg aggacgccaa gctgcaactc tccaaggaca cctacgacga tgacctcgat 3601 aacctcctgg cccagatcgg cgatcaatac gcggacctgt tcctcgctgc caagaacctg 3661 tcggacgcca tcctcctgtc agatatcctc cgcgtgaaca ccgagatcac gaaggctcca 3721 ctctctgcct ccatgatcaa gcgctacgac gagcaccatc aggatctgac cctcctgaag 3781 gcgctggtcc gccaacagct cccggagaag tacaaggaga ttttcttcga tcagtcgaag 3841 aacggctacg ctgggtacat cgacggcggg gcctcacaag aggagttcta caagttcatc 3901 aagccaatcc tggagaagat ggacggcacg gaggagctcc tggtgaagct caacagggag 3961 gacctcctgc ggaagcagag aaccttcgat aacggcagca tcccccacca aatccatctc 4021 ggggagctgc acgccatcct gagaaggcaa gaggacttct accctttcct caaggataac 4081 cgggagaaga tcgagaagat cctgaccttc agaatcccat actacgtcgg ccctctcgcg 4141 cgggggaact caagattcgc ttggatgacc cgcaagtctg aggagaccat cacgccgtgg 4201 aacttcgagg aggtggtgga caagggcgct agcgctcagt cgttcatcga gaggatgacc 4261 aacttcgaca agaacctgcc caacgagaag gtgctcccta agcactcgct cctgtacgag 4321 tacttcaccg tctacaacga gctcacgaag gtgaagtacg tcaccgaggg catgcgcaag 4381 ccagcgttcc tgtccgggga gcagaagaag gctatcgtgg acctcctgtt caagaccaac 4441 cggaaggtca cggttaagca actcaaggag gactacttca agaagatcga gtgcttcgat 4501 tcggtcgaga tcagcggcgt tgaggaccgc ttcaacgcca gcctcgggac ctaccacgat 4561 ctcctgaaga tcatcaagga taaggacttc ctggacaacg aggagaacga ggatatcctg 4621 gaggacatcg tgctgaccct cacgctgttc gaggacaggg agatgatcga ggagcgcctg 4681 aagacgtacg cccatctctt cgatgacaag gtcatgaagc aactcaagcg ccggagatac 4741 accggctggg ggaggctgtc ccgcaagctc atcaacggca tccgggacaa gcagtccggg 4801 aagaccatcc tcgacttcct gaagagcgat ggcttcgcca acaggaactt catgcaactg 4861 atccacgatg acagcctcac cttcaaggag gatatccaaa aggctcaagt gagcggccag 4921 ggggactcgc tgcacgagca tatcgcgaac ctcgctggct cccccgcgat caagaagggc 4981 atcctccaga ccgtgaaggt tgtggacgag ctcgtgaagg tcatgggccg gcacaagcct 5041 gagaacatcg tcatcgagat ggccagagag aaccaaacca cgcagaaggg gcaaaagaac 5101 tctagggagc gcatgaagcg catcgaggag ggcatcaagg agctggggtc ccaaatcctc 5161 aaggagcacc cagtggagaa cacccaactg cagaacgaga agctctacct gtactacctc 5221 cagaacggca gggatatgta cgtggaccaa gagctggata tcaaccgcct cagcgattac 5281 gacgtcgatg ctatcgttcc ccagtctttc ctgaaggatg actccatcga caacaaggtc 5341 ctcaccaggt cggacaagaa ccgcggcaag tcagataacg ttccatctga ggaggtcgtt 5401 aagaagatga agaactactg gaggcagctc ctgaacgcca agctgatcac gcaaaggaag 5461 ttcgacaacc tcaccaaggc tgagagaggc gggctctcag agctggacaa ggccggcttc 5521 atcaagcggc agctggtcga gaccagacaa atcacgaagc acgttgcgca aatcctcgac 5581 tctcggatga acacgaagta cgatgagaac gacaagctga tcagggaggt taaggtgatc 5641 accctgaagt ctaagctcgt ctccgacttc aggaaggatt tccagttcta caaggttcgc 5701 gagatcaaca actaccacca tgcccatgac gcttacctca acgctgtggt cggcaccgct 5761 ctgatcaaga agtacccaaa gctggagtcc gagttcgtgt acggggacta caaggtttac 5821 gatgtgcgca agatgatcgc caagtcggag caagagatcg gcaaggctac cgccaagtac 5881 ttcttctact caaacatcat gaacttcttc aagaccgaga tcacgctggc caacggcgag 5941 atccggaaga gaccgctcat cgagaccaac ggcgagacgg gggagatcgt gtgggacaag 6001 ggcagggatt tcgcgaccgt ccgcaaggtt ctctccatgc cccaggtgaa catcgtcaag 6061 aagaccgagg tccaaacggg cgggttctca aaggagtcta tcctgcctaa gcggaacagc 6121 gacaagctca tcgccagaaa gaaggactgg gacccaaaga agtacggcgg gttcgacagc 6181 cctaccgtgg cctactcggt cctggttgtg gcgaaggttg agaagggcaa gtccaagaag 6241 ctcaagagcg tgaaggagct cctggggatc accatcatgg agaggtccag cttcgagaag 6301 aacccaatcg acttcctgga ggccaagggc tacaaggagg tgaagaagga cctgatcatc 6361 aagctcccga agtactctct cttcgagctg gagaacggca ggaagagaat gctggcttcc 6421 gctggcgagc tccagaaggg gaacgagctc gcgctgccaa gcaagtacgt gaacttcctc 6481 tacctggctt cccactacga gaagctcaag ggcagcccgg aggacaacga gcaaaagcag 6541 ctgttcgtcg agcagcacaa gcattacctc gacgagatca tcgagcaaat ctccgagttc 6601 agcaagcgcg tgatcctcgc cgacgcgaac ctggataagg tcctctccgc ctacaacaag 6661 caccgggaca agcccatcag agagcaagcg gagaacatca tccatctctt caccctgacg 6721 aacctcggcg ctcctgctgc tttcaagtac ttcgacacca cgatcgatcg gaagagatac 6781 acctccacga aggaggtcct ggacgcgacc ctcatccacc agtcgatcac cggcctgtac 6841 gagacgagga tcgacctctc acaactcggc ggggataaga gacccgcagc aaccaagaag 6901 gcagggcaag caaagaagaa gaagacgcgt gactccggcg gcagcaccaa cctgtccgac 6961 atcatcgaga aggagacggg caagcaactc gtgatccagg agagcatcct catgctgcca 7021 gaggaggtgg aggaggtcat cggcaacaag ccagagtccg acatcctggt gcacaccgcc 7081 tacgacgagt ccaccgacga gaacgtcatg ctcctgacca gcgacgcccc agagtacaag 7141 ccatgggccc tcgtcatcca ggacagcaac ggggagaaca agatcaagat gctgtcgggg 7201 gggagcccaa agaagaagcg gaaggtgtag tgtacaagta acccgggaat tcggtacgct 7261 gaaatcacca gtctctctct acaaatctat ctctctctat tttctccata aataatgtgt 7321 gagtagtttc ccgataaggg aaattagggt tcttataggg tttcgctcat gtgttgagca 7381 tataagaaac ccttagtatg tatttgtatt tgtaaaatac ttctatcaat aaaatttcta 7441 attcctaaaa ccaaaatcca gtactaaaat ccagatctcc taaagtccct atagatcttt 7501 gtcgtgaata taaaccagac acgagacgac taaacctgga gcccagacgc cgttcgaagc 7561 tagaagtacc gcttaggcag gaggccgtta gggaaaagat gctaaggcag ggttggttac 7621 gttgactccc ccgtaggttt ggtttaaata tgatgaagtg gacggaagga aggaggaaga 7681 caaggaagga taaggttgca ggccctgtgc aaggtaagaa gatggaaatt tgatagaggt 7741 acgctactat acttatacta tacgctaagg gaatgcttgt atttataccc tataccccct 7801 aataacccct tatcaattta agaaataatc cgcataagcc cccgcttaaa aattggtatc 7861 agagccatga ataggtctat gaccaaaact caagaggata aaacctcacc aaaatacgaa 7921 agagttctta actctaaaga taaaagatct ttcaagatca aaactagttc cctcacaccg 7981 gtgacgggga tcgcatgcga tatctcgaga tctagcttgg cgtaatcatg gtcatagctg 8041 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata 8101 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca 8161 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc 8221 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg 8281 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta 8341 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc 8401 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 8461 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 8521 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 8581 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 8641 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 8701 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 8761 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 8821 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta 8881 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 8941 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg 9001 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag 9061 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc 9121 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact 9181 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt 9241 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta 9301 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta 9361 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc 9421 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat 9481 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt 9541 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg 9601 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca 9661 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta 9721 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg 9781 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact 9841 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg 9901 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt 9961 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga 10021 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc 10081 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa 10141 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgccagtgc caagctaatt 10201 c //