质粒名称: | pHR-FKBP:mCherry-Rab5a |
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出品公司: | addgene |
目录编号: | 72901 |
存储实验室: | Ivan Yudushkin |
载体骨架基本信息 |
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载体名称: | pHR |
空载体大小: | 10644bp |
完整质粒大小: | 10714bp |
载体类型: | 哺乳细胞表达 |
筛选标记: | 新霉素(G418) |
载体抗性: | 氨苄霉素 |
生长温度: | 37 ℃ |
宿主菌: | NEB Stable |
拷贝数: | 高拷贝 |
插入基因信息 |
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插入基因名称: | Rab5a |
物种来源: | H. sapiens (human) |
基因突变: | fragment a.a. 2-215 |
GenBank编号 | NM_004162.4 |
启动子: | CMV |
克隆方法: | 限制性内切酶 |
5’克隆位点: | MluI(未破坏) |
3’克隆位点: | Not (未破坏) |
5’测序引物 | gcttcccgagctctataaaagagc |
3’测序引物 | ccagaggttgattatcgataagc |
文献引用方法 |
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材料和方法部分: | pHR-FKBP:mCherry-Rab5a was a gift from Ivan Yudushkin (Addgene plasmid # 72901 ; http://n2t.net/addgene:72901 ; RRID:Addgene_72901) |
参考文献部分: | Localization of mTORC2 activity inside cells. Ebner M, Sinkovics B, Szczygiel M, Ribeiro DW, Yudushkin I. J Cell Biol. 2017 Jan 31. pii: jcb.201610060. doi: 10.1083/jcb.201610060. 10.1083/jcb.201610060 PubMed 28143890 |
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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 pHR-FKBP:mCherry-Rab5a 10714 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Early endosome-targeted mCherry-tagged recruiter construct. ACCESSION . VERSION . KEYWORDS pHR-FKBP:mCherry-Rab5a SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10714) AUTHORS Ebner M, Sinkovics B, Szczygiel M, Ribeiro DW, Yudushkin I TITLE Localization of mTORC2 activity inside cells. JOURNAL J Cell Biol. 2017 Jan 31. pii: jcb.201610060. doi: 10.1083/jcb.201610060. PUBMED 28143890 REFERENCE 2 (bases 1 to 10714) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..10714 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 89..322 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" promoter complement(347..365) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(348..365) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(655..674) /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind complement(888..907) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 1007..1029 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(1067..1085) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 1153..1257 /gene="bla" /label=AmpR promoter CDS 1258..2118 /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(1476..1495) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2289..2877 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2778..2797 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3031..3048 /label=L4440 /note="L4440 vector, forward primer" primer_bind complement(3118..3138) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3123..3452 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3303..3438 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3365..3384 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" intron 4586..4651 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" CDS 4781..4801 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" polyA_signal 5226..5360 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5263..5282) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5317..5336 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" LTR 5528..6161 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 6208..6333 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 6830..7063 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 7248..7292 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" misc_feature 7577..7694 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 7847..8254 /label=SFFV promoter /note="spleen focus-forming virus long terminal repeat (LTR) promoter" regulatory 8288..8297 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 8294..8617 /codon_start=1 /product="human FK506-binding protein FKBP12" /label=FKBP /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL LKLE" CDS 8642..9349 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(8787..8805) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 9055..9074 /label=mCherry-F /note="mCherry, forward primer" regulatory 9359..9368 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 9371..9397 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" misc_feature 10105..10693 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(10158..10178) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(10576..10587) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(10696..10712) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(10697..10713) /label=pBluescriptKS /note="For pBluescript vector" ORIGIN 1 ggaattaatt cgagctcggt acctttaaga ccaatgactt acaaggcagc tgtagatctt 61 agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa 121 gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc 181 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa 241 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag 301 tcagtgtgga aaatctctag cagcatctag aattaattcc gtgtattcta tagtgtcacc 361 taaatcgtat gtgtatgata cataaggtta tgtattaatt gtagccgcgt tctaacgaca 421 atatgtacaa gcctaattgt gtagcatctg gcttactgaa gcagacccta tcatctctct 481 cgtaaactgc cgtcagagtc ggtttggttg gacgaacctt ctgagtttct ggtaacgccg 541 tcccgcaccc ggaaatggtc agcgaaccaa tcagcagggt catcgctagc cagatcctct 601 acgccggacg catcgtggcc ggcatcaccg gcgccacagg tgcggttgct ggcgcctata 661 tcgccgacat caccgatggg gaagatcggg ctcgccactt cgggctcatg agcgcttgtt 721 tcggcgtggg tatggtggca ggccccgtgg ccgggggact gttgggcgcc atctccttgc 781 atgcaccatt ccttgcggcg gcggtgctca acggcctcaa cctactactg ggctgcttcc 841 taatgcagga gtcgcataag ggagagcgtc gaatggtgca ctctcagtac aatctgctct 901 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg 961 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg 1021 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc 1081 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt 1141 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat 1201 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg 1261 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt 1321 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga 1381 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa 1441 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt 1501 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt 1561 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc 1621 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga 1681 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat 1741 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct 1801 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc 1861 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg 1921 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc 1981 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg 2041 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca 2101 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta 2161 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc 2221 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa 2281 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca 2341 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta 2401 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc 2461 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca 2521 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta 2581 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag 2641 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt 2701 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc 2761 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac 2821 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac 2881 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc 2941 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat 3001 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag 3061 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctgtgga 3121 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 3181 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca 3241 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc 3301 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt 3361 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag 3421 gaggcttttt tggaggccta ggcttttgca aaaagcttgg acacaagaca ggcttgcgag 3481 atatgtttga gaataccact ttatcccgcg tcagggagag gcagtgcgta aaaagacgcg 3541 gactcatgtg aaatactggt ttttagtgcg ccagatctct ataatctcgc gcaacctatt 3601 ttcccctcga acacttttta agccgtagat aaacaggctg ggacacttca catgagcgaa 3661 aaatacatcg tcacctggga catgttgcag atccatgcac gtaaactcgc aagccgactg 3721 atgccttctg aacaatggaa aggcattatt gccgtaagcc gtggcggtct gtaccgggtg 3781 cgttactggc gcgtgaactg ggtattcgtc atgtcgatac cgtttgtatt tccagctacg 3841 atcacgacaa ccagcgcgag cttaaagtgc tgaaacgcgc agaaggcgat ggcgaaggct 3901 tcatcgttat tgatgacctg gtggataccg gtggtactgc ggttgcgatt cgtgaaatgt 3961 atccaaaagc gcactttgtc accatcttcg caaaaccggc tggtcgtccg ctggttgatg 4021 actatgttgt tgatatcccg caagatacct ggattgaaca gccgtgggat atgggcgtcg 4081 tattcgtccc gccaatctcc ggtcgctaat cttttcaacg cctggcactg ccgggcgttg 4141 ttctttttaa cttcaggcgg gttacaatag tttccagtaa gtattctgga ggctgcatcc 4201 atgacacagg caaacctgag cgaaaccctg ttcaaacccc gctttaaaca tcctgaaacc 4261 tcgacgctag tccgccgctt taatcacggc gcacaaccgc ctgtgcagtc ggcccttgat 4321 ggtaaaacca tccctcactg gtatcgcatg attaaccgtc tgatgtggat ctggcgcggc 4381 attgacccac gcgaaatcct cgacgtccag gcacgtattg tgatgagcga tgccgaacgt 4441 accgacgatg atttatacga tacggtgatt ggctaccgtg gcggcaactg gatttatgag 4501 tgggccccgg atctttgtga aggaacctta cttctgtggt gtgacataat tggacaaact 4561 acctacagag atttaaagct ctaaggtaaa tataaaattt ttaagtgtat aatgtgttaa 4621 actactgatt ctaattgttt gtgtatttta gattccaacc tatggaactg atgaatggga 4681 gcagtggtgg aatgccttta atgaggaaaa cctgttttgc tcagaagaaa tgccatctag 4741 tgatgatgag gctactgctg actctcaaca ttctactcct ccaaaaaaga agagaaaggt 4801 agaagacccc aaggactttc cttcagaatt gctaagtttt ttgagtcatg ctgtgtttag 4861 taatagaact cttgcttgct ttgctattta caccacaaag gaaaaagctg cactgctata 4921 caagaaaatt atggaaaaat attctgtaac ctttataagt aggcataaca gttataatca 4981 taacatactg ttttttctta ctccacacag gcatagagtg tctgctatta ataactatgc 5041 tcaaaaattg tgtaccttta gctttttaat ttgtaaaggg gttaataagg aatatttgat 5101 gtatagtgcc ttgactagag atcataatca gccataccac atttgtagag gttttacttg 5161 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg 5221 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt 5281 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg 5341 tatcttatca tgtctggatc aactggataa ctcaagctaa ccaaaatcat cccaaacttc 5401 ccaccccata ccctattacc actgccaatt acctgtggtt tcatttactc taaacctgtg 5461 attcctctga attattttca ttttaaagaa attgtatttg ttaaatatgt actacaaact 5521 tagtagttgg aagggctaat tcactcccaa agaagacaag atatccttga tctgtggatc 5581 taccacacac aaggctactt ccctgattag cagaactaca caccagggcc aggggtcaga 5641 tatccactga cctttggatg gtgctacaag ctagtaccag ttgagccaga taaggtagaa 5701 gaggccaata aaggagagaa caccagcttg ttacaccctg tgagcctgca tgggatggat 5761 gacccggaga gagaagtgtt agagtggagg tttgacagcc gcctagcatt tcatcacgtg 5821 gcccgagagc tgcatccgga gtacttcaag aactgctgat atcgagcttg ctacaaggga 5881 ctttccgctg gggactttcc agggaggcgt ggcctgggcg ggactgggga gtggcgagcc 5941 ctcagatcct gcatataagc agctgctttt tgcctgtact gggtctctct ggttagacca 6001 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag 6061 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag 6121 atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac 6181 ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg 6241 cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga 6301 ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg 6361 cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata 6421 gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca 6481 gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa 6541 cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata 6601 aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc 6661 gcacagcaag cggccggccg ctgatcttca gacctggagg aggagatatg agggacaatt 6721 ggagaagtga attatataaa tataaagtag taaaaattga accattagga gtagcaccca 6781 ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt 6841 tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg 6901 tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta 6961 ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa 7021 gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct 7081 ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc 7141 tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca 7201 caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag 7261 aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc 7321 tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt 7381 ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga 7441 cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag 7501 agagagacag agacagatcc attcgattag tgaacggatc tcgacggtat cgccaaatgg 7561 cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa 7621 gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa 7681 aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccagtttgg atcgataagc 7741 ttgatatcga attcctgcag ccccgataaa ataaaagatt ttatttagtc tccagaaaaa 7801 ggggggaatg aaagacccca cctgtaggtt tggcaagcta gctgcagtaa cgccattttg 7861 caaggcatgg aaaaatacca aaccaagaat agagaagttc agatcaaggg cgggtacatg 7921 aaaatagcta acgttgggcc aaacaggata tctgcggtga gcagtttcgg ccccggcccg 7981 gggccaagaa cagatggtca ccgcagtttc ggccccggcc cgaggccaag aacagatggt 8041 ccccagatat ggcccaaccc tcagcagttt cttaagaccc atcagatgtt tccaggctcc 8101 cccaaggacc tgaaatgacc ctgcgcctta tttgaattaa ccaatcagcc tgcttctcgc 8161 ttctgttcgc gcgcttctgc ttcccgagct ctataaaaga gctcacaacc cctcactcgg 8221 cgcgccagtc ctccgacaga ctgagtcgcc cgggggggat ctggagctct cgagaattct 8281 cacgcgtgcc accatgggag tgcaggtgga gactatctcc ccaggagacg ggcgcacctt 8341 ccccaagcgc ggccagacct gcgtggtgca ctacaccggg atgcttgaag atggaaagaa 8401 atttgattcc tcccgggaca gaaacaagcc ctttaagttt atgctaggca agcaggaggt 8461 gatccgaggc tgggaagaag gggttgccca gatgagtgtg ggtcagagag ccaaactgac 8521 tatatctcca gattatgcct atggtgccac tgggcaccca ggcatcatcc caccacatgc 8581 cactctcgtc ttcgatgtgg agcttctaaa actggaaggt gctgcacttc tgcccggttc 8641 tatggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa 8701 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg 8761 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc 8821 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca 8881 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg 8941 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga 9001 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt 9061 aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg 9121 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc 9181 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt 9241 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga 9301 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagc ccggttctgc 9361 cgccatggca tatccatatg acgtcccaga ctatgctggc gctagtcgag gcgcaacaag 9421 acccaacggg ccaaatactg gaaataaaat atgccagttc aaactagtac ttctgggaga 9481 gtctgctgtt ggcaaatcaa gcctagtgct tcgttttgtg aagggccaat ttcatgaatt 9541 tcaagagagt accatagggg ctgcttttct aacccaaact gtgtgtcttg atgatacaac 9601 agtaaagttt gaaatatggg atacagctgg tcaagaacga taccatagct tagcaccaat 9661 gtactacaga ggagcacaag cagccatagt tgtatatgat atcacaaatg aggagtcctt 9721 tgccagagcc aaaaactggg ttaaagaact tcagaggcaa gccagtccta acattgtaat 9781 agctttatca ggaaacaagg ctgatcttgc aaataaaaga gctgtcgatt tccaggaagc 9841 acagtcctat gcagatgaca acagtttatt attcatggag acatcagcta aaacatcgat 9901 gaatgtaaat gaaatattca tggcaatagc taaaaagttg ccaaagaacg aaccacagaa 9961 tccaggagca aattctgcca gaggaagagg agtagacctt actgaaccca cgcagccaac 10021 caggaatcag tgttgtagta actaagcggc cgcgactcta gagtcgacct gcaggcatgc 10081 aagcttgata tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg 10141 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct 10201 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg 10261 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact 10321 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc 10381 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc 10441 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa 10501 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc 10561 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg 10621 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg 10681 gccgcctccc cgcatcgata ccgtcgacct cgag //
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