质粒名称: | pcDNA3.1-hAsCpf1(TYCV) (pY210) |
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出品公司: | Addgene |
目录编号: | 89351 |
存储实验室: | Feng Zhang |
载体骨架基本信息 |
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载体名称: | pcDNA3.1 |
载体抗性: | 氨苄青霉素 |
空载体大小: | 5402 bp |
完整质粒大小: | 9509 bp |
载体类型: | 哺乳细胞表达,CRISPR |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | Acidaminococcus sp. Cpf1 (RR variant) |
物种来源: | Synthetic |
插入大小: | 4107 bp |
基因突变: | S542R/K607R |
启动子 | CMV |
标签/融合蛋白: |
NLS(插入件上的N端子)
NLS(插入件上的C端子)
3xHA(插入件上的C端子)
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克隆方法: | Gibson克隆 |
5' 测序引物 | CGCAAATGGGCGGTAGGCGTG |
文献引用方法 |
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材料和方法部分: | pcDNA3.1-hAsCpf1(TYCV) (pY210) was a gift from Feng Zhang (Addgene plasmid # 89351 ; http://n2t.net/addgene:89351 ; RRID:Addgene_89351) |
参考文献部分: | Engineered Cpf1 variants with altered PAM specificities. Gao L, Cox DBT, Yan WX, Manteiga JC, Schneider MW, Yamano T, Nishimasu H, Nureki O, Crosetto N, Zhang F. Nat Biotechnol. 2017 Jun 5. doi: 10.1038/nbt.3900. 10.1038/nbt.3900 PubMed 28581492 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pcDNA3.1-hAsCpf1(TYCV) (pY210) 9509 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION Expresses humanized AsCpf1 variant that recognizes TYCV PAMs. ACCESSION . VERSION . KEYWORDS pcDNA3.1-hAsCpf1(TYCV) (pY210) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9509) AUTHORS Gao L, Cox DBT, Yan WX, Manteiga JC, Schneider MW, Yamano T, Nishimasu H, Nureki O, Crosetto N, Zhang F TITLE Engineered Cpf1 variants with altered PAM specificities. JOURNAL Nat Biotechnol. 2017 Jun 5. doi: 10.1038/nbt.3900. PUBMED 28581492 REFERENCE 2 (bases 1 to 9509) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9509 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 213..233 /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin complement(242..670) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(352..373) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 564..583 /label=F1ori-R /note="F1 origin, reverse primer" polyA_signal 716..940 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind 929..946 /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" CDS complement(1020..1100) /codon_start=1 /product="three tandem HA epitope tags" /label=3xHA /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA" CDS complement(1107..1154) /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" CDS complement(1155..5072) /codon_start=1 /product="CRISPR-associated protein from Acidaminococcus sp. BV3L6 (Zetsche et al., 2015)" /label=AsCpf1 /translation="TQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARNDH YKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRNA IHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFDK FTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHFE NVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVLN LAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLLR NENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTGK ITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTLK KQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKAR NYATKKPYSVEKFKLNFQMPTLARGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYKA LSFEPTEKTSEGFDKMYYDYFPDAAKMIPRCSTQLKAVTAHFQTHTTPILLSNNFIEPL EITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSIDL SSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKGH HGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLKD QKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFFH VPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQRS LNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAVV VLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQF TSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYDV KTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIENH RFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQMR NSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKESK DLKLQNGISNQDWLAYIQELRN" regulatory 1426..1435 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS complement(5097..5117) /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" regulatory 5120..5129 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" primer_bind complement(5164..5183) /label=T7 /note="T7 promoter, forward primer" promoter complement(5165..5183) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" promoter complement(5228..5431) /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind complement(5257..5277) /label=CMV-F /note="Human CMV immediate early promoter, forward primer" enhancer 5432..5811 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" primer_bind 5983..6002 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 6077..6181 /gene="bla" /label=AmpR promoter CDS 6182..7042 /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(6400..6419) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7213..7801 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7702..7721 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 7955..7972 /label=L4440 /note="L4440 vector, forward primer" protein_bind 8089..8110 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 8125..8155 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8163..8179 /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 8168..8190 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 8187..8203 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 8187..8203 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" polyA_signal 8252..8373 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(8263..8282) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 8317..8336 /label=SV40pA-R /note="SV40 polyA, reverse primer" CDS complement(8547..9341) /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(8658..8677) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 9268..9287 /label=Neo-R /note="Neomycin resistance gene, reverse primer" promoter complement(9408..228) /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 9422..48 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(9476..9495) /label=SV40pro-F /note="SV40 promoter/origin, forward primer" ORIGIN 1 gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag 61 gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg 121 ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc tttgcatact 181 tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat tccacagaat 241 taattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc 301 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg 361 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat 421 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc 481 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag 541 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg 601 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta 661 cagggcgcgt ggggataccc cctagagccc cagctggttc tttccgcctc agaagccata 721 gagcccaccg catccccagc atgcctgcta ttgtcttccc aatcctcccc cttgctgtcc 781 tgccccaccc caccccccag aatagaatga cacctactca gacaatgcga tgcaatttcc 841 tcattttatt aggaaaggac agtgggagtg gcaccttcca gggtcaagga aggcacgggg 901 gaggggcaaa caacagatgg ctggcaacta gaaggcacag tcgaggctga tcagcgggtt 961 taaacgggcc ctctagactc gagcggccgc cactgtgctg gatatctgca gaattcttag 1021 gcatagtcgg ggacatcata tgggtatgca taatcaggca cgtcgtaggg ataagcgtaa 1081 tctggaacat cgtatgggta ggatcccttt ttcttttttg cctggccggc ctttttcgtg 1141 gccgccggcc ttttgttgcg cagctcctgg atgtaggcca gccagtcctg attggagatg 1201 ccgttctgca gcttcagatc cttgctctcc ttcaggtgat tcagcagcag ctggcccttc 1261 agggcgatgt ggtaggcgcc attggcatcg gcgtccatgg gccactctgg gttctgaaac 1321 cgggagtcga agcacacgcc attcagatcg cgcacggggc tgttgatata gtcctcgcct 1381 gtggcggcat tggagttccg catctgcagc acgctgcgga tcagggccac catggtgtcg 1441 atggcgtgag aatcgtcatt ctccagcagc tttggcagga tgttggagcc atccctgaac 1501 acgatgccct tctcctccag cagggcgatc agctcgttgg caggatacag gtcccggtat 1561 ctgccggtga atctgtgatt ctcgatcact ggcacgattc tcttgccggc gatgaaaggg 1621 gtgcccttgg cgtcaaactg tgtctcgttc ttctcgaaca cgatatccca tgcaggcata 1681 aagccgggca ggcccctctg gaaggacaga tttctgttca tcttaaagtg caggatgaag 1741 tcgccggttt tcacgtcgta gtgcagaaag tcgaagccct ccaggaagtg cttgcggctc 1801 tcgtgattct tgatggtttt ccacacgaag gggtccacga agccggtcag gggatcgatc 1861 ttagatgtat atggggcagg cacgtaaaac aggaagccag actgggtgcc catcttggca 1921 aaggaggtga actggtctgt cagctggtat gggttcagca cgcctcccac tttctctgct 1981 ggatagtcct tcagcaccag gcaattcagc ttatcgatca gcatcttctc gaactgctgg 2041 tacacggcct tctcggcgat gccggtcctc ttgctcttaa agccgaaatt caggttctcc 2101 agcaccacca cggcctggta gtggatcatc aggtccacga tctcgtggat gacctggctc 2161 agatagccct gcttcagatc cttgattgtg cccaccacag accaggcctg ccttgctgcc 2221 accctctcct tctccctgtt gtccagcttc ttctggtaat caaactgctg gatggtgttc 2281 aggctccgct gctccaggat cttgccggtg gagtcgatca ctgtgatata gatcaggttt 2341 ctctcgcccc gatcgatgcc gatgataggt gtctcggggt gctccttcag gtaggcattc 2401 accctctggt tgaacttaga tggggaattg gcggcctgat agttcagtgt gataggcacg 2461 tggaaaaaga acttgtcgct ggtaaagcgc ctatccttga tgatctcgtg agacacctcc 2521 ttggtgatca cgttgggcag cagggccctg gcctcatcag acaggtcgtg ggacagtctg 2581 tgattcacat agtcgtacag ctcctggtac agggtgtcgg ggattggggt tttctgatcc 2641 ttcagcttct tgttcagcat cttctctccc agccggtgtg ccatcctctt catcctggac 2701 ttagggcggt agaacagctc ggcctggcca ttcagcttga tgcttgtctt ggccaggttc 2761 tctggagaaa acagaccggt ccaatacagt gtgtgcagat taggcttgcc gtggtggccc 2821 ttggcaaagt ccttgttata gatctggaac aggtacagct tgcctgtctc cacggcatcc 2881 atgatctcct tctcggcgat tctctggaag ctgatgtggt acagcagggg attcagctcg 2941 gcatagtact cgcccaggtc cttatactga gaggatggcc gcaggctaga cagatcgata 3001 gaggttgtct tggtatactt ggacagaaaa tcccttgtga agtcgatcca cttgcacagg 3061 gcctctctgt agcccttctg gtcgccggtt ttcttggcgt acgctgtctg aaacttcttt 3121 ggctccttct caggattgtt caggtcgtag atctcctttg tgatctccag aggctcgatg 3181 aaattgttgg acagcaggat gggggttgtg tgggtctgaa agtgggcggt caccgccttc 3241 agctgggtgc tgcaccgtgg gatcatcttg gcggcatccg gaaagtagtc atagtacatc 3301 ttatcaaagc cctcgctggt tttctctgtg ggctcgaagc tgagggcctt atacctgccc 3361 ttctgctttg gcatgatgcc cagatagtac aggccgttct tcacaaacag gatggcgcca 3421 ttgttcttct ccttattcac gtcccagccc cgggccagtg taggcatctg aaagttcagc 3481 ttgaacttct ccacggagta gggcttcttg gtggcataat ttctggcctt gttgtagaag 3541 ctcagagaag gctccatctc cagcttgatg ccggtcagcc gggcagagaa ctcggggtcc 3601 acctcgttgg actcatccac ggcaaaccag tccagcaggt ggtacaggcc cagcaggctg 3661 tccagctgag acttcaggat ctccttctcc tcctgcttct tcagggttgt aggcagtggc 3721 tgatccaggg cggcgtgtgc gtgggacagg atctcgctgg ttttctgctt gaaggcctcg 3781 ctcagctcct tgcctgcggc agagatgatc tcctgcaggt tgatatcctc gtgcttcagg 3841 ctgcgctgca ccttctcctt ggcagacttg gtgatcttgc ctgtcagctc ggagattctc 3901 cgctcataca gggcattcct cagtgtatcc cagtggtcgc acagggcgct gctgattgtc 3961 tccagcttct tgtggctgat gaagatgtgt gtcaggtcga tgctgttcag ctcgttaaac 4021 agggcctcgg ctgtctccag cacgttctcg tttctcagca gtgtcttgta cttgcagaag 4081 gactggatca cttcctcgtc gctcttaaac tcctccagga tgaaagacag ggtgttccta 4141 tcggacagga tctgcttaaa cagggggatg aatctgtgtg gcagggaggc gatgatgtgg 4201 gctgtctcat cattcttctg gatggccaga ttcagcacct cgttcaggcc cttgatcttc 4261 tcggtgcctg cctcccgaga gattcctccc agcagctggt tatacaggtc gatctgggtc 4321 tgtgtcagca gctggttata aaaagggaag gaaaacacct cctcgatgga ggtgctcacg 4381 aagatgccga tggccttctt cacgttctca aagtgctccc gcaggctggg cacggcggtg 4441 atcaggcgtg tgaagatgtg acaattctcc ttaaacttgg ggaagttgtc ctgcacgatg 4501 cggtgtggga tggctgtgct gatatcctcg gcgctgaaca cgttcttcct gttctcataa 4561 aagccggaga agtaggttgt aaacttgtcg aagctccgca gcagggcgtt ctcgtgctcg 4621 gttgtggtca cggtgcccag ctgcttcagc accttgccat taaacagctc ggccttgaac 4681 aggcccttgt agatctcggc gtgtctctta ttgatggcat cggtcaggtt gtctgtccgg 4741 ccgatgaagt agtcgtggat ggcattgcga tatgtggcct gctcctcgat cagggcgttc 4801 cttgtctcct cggttttctc ctttctatag gagtcgatgg cggcgctcag gttctcccaa 4861 tccagctgca ccagctgcag gcactggtcg gcataggtct tgtagatccg atcgatgatg 4921 ggcttcagct ccttgtagtg atcattgcgg gccttgtcct cctcgatgaa gccctgctcc 4981 tggatgtgct tcagggtctt gccctgtggg atcagctcaa accgcagtgt cttgctcacc 5041 tgatacaggt tggtaaagcc ctcgaactgt gtggctgctg ggactccgtg gataccgacc 5101 ttccgcttct tctttggggc catggtggca agcttaagtt taaacgctag ccagcttggg 5161 tctccctata gtgagtcgta ttaatttcga taagccagta agcagtgggt tctctagtta 5221 gccagagagc tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca 5281 atggggcgga gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac 5341 tcccattgac gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac 5401 gcccattgat gtactgccaa aaccgcatca ccatggtaat agcgatgact aatacgtaga 5461 tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc 5521 atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg 5581 ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg 5641 gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc 5701 agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa 5761 ctaatgaccc cgtaattgat tactattaat aactagtcaa taatcaatgt caacgcgtat 5821 atctggcccg tacatcgcga agcagcgcaa aacgcctaac cctaagcaga ttcttcatgc 5881 aattgtcggt caagccttgc cttgttgtag cttaaatttt gctcgcgcac tactcagcga 5941 cctccaacac acaagcaggg agcagatact ggcttaacta tgcggcatca gagcagattg 6001 tactgagagt gcaccatagg ggatcgggag atctcccgat ccgtcgacgt caggtggcac 6061 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat 6121 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag 6181 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc 6241 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc 6301 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc 6361 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc 6421 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt 6481 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt 6541 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat 6601 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct 6661 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat 6721 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc 6781 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg 6841 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc 6901 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta 6961 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc 7021 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga 7081 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat 7141 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat 7201 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa 7261 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa 7321 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt 7381 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt 7441 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata 7501 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt 7561 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac 7621 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga 7681 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg 7741 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa 7801 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat 7861 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc 7921 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga 7981 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg 8041 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta 8101 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg 8161 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct 8221 ctagctagag gtcgacggta tacagacatg ataagataca ttgatgagtt tggacaaacc 8281 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta 8341 tttgtaacca ttataagctg caataaacaa gttggggtgg gcgaagaact ccagcatgag 8401 atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct 8461 ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc 8521 ggtcatttcg aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg 8581 atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg 8641 ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc 8701 acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc 8761 ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gcgcgccttg 8821 agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga 8881 tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg 8941 tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg 9001 gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc 9061 aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg 9121 cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct gcagttcatt cagggcaccg 9181 gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg 9241 gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa 9301 gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct 9361 gtctcttgat cagatccgaa aatggatata caagctcccg ggagcttttt gcaaaagcct 9421 aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggcaga ggcggcctcg 9481 gcctctgcat aaataaaaaa aattagtca //