质粒名称: | pcDNA5-T/O-Ds1-mCherry |
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出品公司: | Addgene |
目录编号: | 112855 |
存储实验室: | David Sprinzak |
载体骨架基本信息 |
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载体名称: | pcDNA5/TO |
载体出品公司: | Thermofisher |
载体类型: | 哺乳细胞表达 |
筛选标记: | 潮霉素 |
载体抗性: | 氨苄霉素 |
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: | DCHS1 |
基因又名: | dachsous cadherin-related 1, DCHS1 (a.k.a. CDH19, CDH25, CDHR6, FIB1, MVP2, PCDH16, VMLDS1) |
物种来源: | H. sapiens (human) |
启动子: | doxycycline inducible promoter |
标签/融合蛋白: | mCherry (C terminal on insert) |
克隆方法: | 限制性内切酶 |
5’克隆位点: | NotI(未破坏) |
3’克隆位点: | AgeI (未破坏) |
5’测序引物 | CMV-F |
文献引用方法 |
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材料和方法部分: | pcDNA5-T/O-Ds1-mCherry was a gift from David Sprinzak (Addgene plasmid # 112855 ; http://n2t.net/addgene:112855 ; RRID:Addgene_112855) |
参考文献部分: | A synthetic planar cell polarity system reveals localized feedback on Fat4-Ds1 complexes. Loza O, Heemskerk I, Gordon-Bar N, Amir-Zilberstein L, Jung Y, Sprinzak D. Elife. 2017 Aug 18;6. pii: 24820. doi: 10.7554/eLife.24820. 24820 [pii] PubMed 28826487 |
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载体基本信息
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载体简介
载体序列
LOCUS pcDNA5-T/O-Ds1-mCherry 16256 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Dox inducible expression in mammalian cells of Ds1 protein fused to mCherry fluorescent protein. ACCESSION . VERSION . KEYWORDS pcDNA5-T/O-Ds1-mCherry SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 16256) AUTHORS Loza O, Heemskerk I, Gordon-Bar N, Amir-Zilberstein L, Jung Y, Sprinzak D TITLE A synthetic planar cell polarity system reveals localized feedback on Fat4-Ds1 complexes. JOURNAL Elife. 2017 Aug 18;6. pii: 24820. doi: 10.7554/eLife.24820. PUBMED 28826487 REFERENCE 2 (bases 1 to 16256) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..16256 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 381..584 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 535..555 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" protein_bind 586..604 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 607..625 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 635..659 /label=LNCX /note="Human CMV promoter, forward primer" CDS 10693..11406 /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 EGRHSTGGMDELYKS" primer_bind complement(10838..10856) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 11106..11125 /label=mCherry-F /note="mCherry, forward primer" primer_bind complement(11444..11461) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 11450..11674 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 11720..12148 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(11807..11826) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 12017..12038 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(12157..12177) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 12162..12491 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 12342..12477 /label=SV40 ori /note="SV40 origin of replication" primer_bind 12404..12423 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 12540..13565 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK E" polyA_signal 13695..13816 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(13732..13751) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 13786..13805 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(13865..13881) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(13865..13881) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(13878..13900) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 13889..13905 /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(13913..13943) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 13958..13979 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(14096..14113) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(14267..14855) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(14347..14366) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(15026..15886) /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 15649..15668 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(15887..15991) /gene="bla" /label=AmpR promoter primer_bind complement(16066..16085) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc 421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt 481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa 541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctccct atcagtgata 601 gagatctccc tatcagtgat agagatcgtc gacgagctcg tttagtgaac cgtcagatcg 661 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc 721 tccggactct agcgtttaaa cttaagcttg gtaccgagct cggatccgcg gccgctcgcc 781 accatgcaga aggagctggg cattgtgcct tcctgccctg gcatgaagag ccccaggccc 841 cacctcctgc taccattgct gctgctgctg ctgctgctgc tgggggctgg ggtgccaggt 901 gcctggggtc aggctgggag cctggacttg cagattgatg aggagcagcc agcgggtaca 961 ctgattggcg acatcagtgc ggggcttccg gcaggcacgg cagctcctct catgtacttc 1021 atctctgccc aagagggcag cggcgtgggc acagacctgg ccattgacga acacagtggg 1081 gtcgtccgta cagcccgtgt cttggaccgt gagcagcggg accgctaccg cttcactgca 1141 gtcactcctg atggtgccac cgtagaagtt acagtgcgag tggctgacat caacgaccat 1201 gctccagcct tcccacaggc tcgggctgcc ctgcaggtac ctgagcatac agcttttggc 1261 acccgctacc cactggagcc tgctcgtgat gcagatgctg ggcgtctggg aacccagggc 1321 tatgcgctat ctggtgatgg ggctggagag accttccggc tggagacacg ccccggtcca 1381 gatgggactc cagtacctga gctggtagtt actggggaac tggaccgaga gaaccgctca 1441 cactatatgc tacagctgga ggcctatgat ggtggttcac ccccccggag ggcccaggcc 1501 ctgctggacg tgacactgct ggacatcaat gaccatgccc cggctttcaa tcagagccgc 1561 taccatgctg tggtgtctga gagcctggcc cctggcagtc ctgtcttgca ggtgttcgca 1621 tctgatgccg atgctggtgt caatggggct gtgacttacg agatcaaccg gaggcagagc 1681 gagggtgatg gacccttctc catcgacgca cacacggggc tgctgcagtt agagcggcca 1741 ctggactttg agcagcggcg ggtccatgaa ctggtggtgc aagcacgaga tggtggggct 1801 caccctgagc tgggctcggc ctttgtgact gtgcatgtgc gagatgccaa tgacaatcag 1861 ccctccatga ctgtcatctt tctcagtgca gatggctccc cccaagtgtc tgaggccgcc 1921 ccacctggac agctcgttgc tcgcatctct gtgtcagacc cagatgatgg tgactttgcc 1981 catgtcaatg tgtccctgga aggtggagag ggccactttg ccctaagcac ccaagacagc 2041 gtcatctatc tggtgtgtgt ggctcggcgg ctggatcgag aggagaggga tgcctataac 2101 ttgagggtta cagccacaga ctcaggctca cctccactgc gggctgaggc tgcctttgtg 2161 ctgcacgtca ctgatgtcaa cgacaatgca cctgcctttg accgccagct ctaccgacct 2221 gagcccctgc ctgaggttgc gctgcctggc agctttgtag tgcgggtgac tgctcgggat 2281 cctgaccaag gcaccaatgg tcaggtcact tatagcctag cccctggcgc ccacacccac 2341 tggttctcca ttgaccccac ctcaggcatt atcactacgg ctgcctcact ggactatgag 2401 ttggaacctc agccacagct gattgtggtg gccacagatg gtggcctgcc ccctctagcc 2461 tcctctgcca cagttagcgt ggccctgcaa gatgtgaatg ataatgagcc ccaattccag 2521 aggactttct acaatgcctc actgcctgag ggcacccagc ctggaacttg cttcctgcag 2581 gtgacagcca cagacgcgga tagtggccca tttggcctcc tctcctattc cttgggtgct 2641 ggacttgggt cctccggatc tcccccattc cgcattgatg cccatagcgg tgatgtgtgc 2701 acaacccgga ccctggaccg tgaccagggg ccctcaagct ttgacttcac agtgacagct 2761 gtggatgggg gaggcctcaa gtccatggta tatgtgaagg tgtttctgtc agacgagaat 2821 gacaaccctc ctcagtttta tccacgggag tatgctgcca gtataagtgc ccagagtcca 2881 ccaggcacag ctgtgctgag gttgcgtgcc catgaccctg accagggatc ccatgggcga 2941 ctctcctacc atatcctggc tggcaacagc cccccacttt ttaccttgga tgagcaatca 3001 gggctgttga cagtagcctg gcccttggcc agacgggcca attctgtggt gcagctggag 3061 attggggctg aggacggagg tggcctacag gcagaaccca gtgcccgagt ggacatcagc 3121 attgtgcctg gaacccccac accacccata tttgagcaac tacagtatgt tttttctgtg 3181 ccagaggatg tggcaccagg caccagtgtg ggcatagtcc aggcacacaa cccaccaggt 3241 cgcttggcac ctgtgaccct ttccctatca ggtggggatc cccgaggact cttctcccta 3301 gatgcggtat caggactgtt gcaaacactt cgccctctgg accgggagct actgggacca 3361 gtgttggagc tggaggtgcg agcaggcagt ggagtgcccc cagctttcgc tgtagctcgg 3421 gtgcgtgtgc tgctggatga tgtgaatgac aactcccctg cctttcctgc acctgaagac 3481 acggtattgc taccaccaaa cactgcccca gggactccca tctatacact gcgggctctt 3541 gaccccgact caggtgttaa cagtcgagtc acctttaccc tgcttgctgg gggtggtgga 3601 gccttcaccg tggaccccac cacaggccat gtacggctta tgaggcctct ggggccctca 3661 ggagggccag cccatgagct ggagctggag gcccgggatg ggggctcccc accacgcacc 3721 agccactttc gactacgggt ggtggtacag gatgtgggaa cccgtgggct ggctccccga 3781 ttcaacagcc ctacctaccg tgtggacctg ccctcaggca ccactgctgg aactcaggtc 3841 ctgcaagtgc aggcccaagc accagatggg ggccctatca cctatcacct tgcagcagag 3901 ggagcaagta gcccctttgg cctggagcca cagagtgggt ggctatgggt gcgggcagca 3961 ctagaccgtg aggcccagga attgtacata ctgaaggtaa tggcagtgtc tgggtccaaa 4021 gctgagttgg ggcagcagac aggcacagcc accgtgaggg tcagcatcct caaccagaat 4081 gaacacagtc cccgcttgtc tgaggatccc accttcctgg ctgtggctga gaaccagccc 4141 ccagggacca gcgtgggccg agtctttgcc actgaccgag actcaggacc caatggacgt 4201 ctgacctaca gcctgcaaca gctgtctgaa gacagcaagg ccttccgcat ccacccccag 4261 actggagaag tgaccacact ccaaaccctg gaccgtgagc agcagagcag ctatcagctc 4321 ctggtgcagg tgcaggatgg agggagccca ccccgcagca ccacaggcac tgtgcatgtt 4381 gcagtgcttg acctcaacga caacagcccc acgttcctgc aggcttcagg agctgctggt 4441 gggggcctcc ctatacaggt accagaccgc gtgcctccag gaacactggt gacgactctg 4501 caggcgaagg atccagatga gggggagaat gggaccatct tgtacacgct aactggtcct 4561 ggctcagagc ttttctctct gcaccctcac tcaggggagc tgctcactgc agctcccctg 4621 atccgagcag agcggcccca ctatgtgctg acactgagtg ctcatgacca aggcagccct 4681 cctcgaagtg ccagcctcca gctgctggtg caggtgcttc cctcagctcg cttggccgag 4741 ccgcccccag atctcgcaga gcgggaccca gcggcaccag tgcctgtcgt gctgacggtg 4801 acagcagctg agggactgcg gcccggctct ctgttgggct cggtggcagc gccagagccc 4861 gcgggtgtgg gtgcactcac ctacacactg gtgggcggtg ccgatcccga gggcaccttc 4921 gcgctggatg cggcctcagg gcgcttgtac ctggcgcggc ccctggactt cgaagctggc 4981 ccgccgtggc gcgcgctcac ggtacgcgct gaggggccgg gaggcgcggg cgcgcggctg 5041 ctgcgagtgc aggtgcaagt gcaggacgag aatgagcatg cgcccgcctt tgcgcgcgac 5101 ccgctggcgc tggcgctgcc agagaacccg gagcccggcg cagcgctgta cactttccgc 5161 gcgtcggacg ccgacggccc cggccccaat agcgacgtgc gctaccgcct gctgcgccag 5221 gagccgcccg tgccggcgct tcgcctggac gcgcgcaccg gggcgctcag cgctccgcgc 5281 ggcctggacc gagagaccac tcccgcgctg ctgctgctgg tggaagccac cgaccggccc 5341 gccaacgcca gccgccgtcg tgcagcgcgc gtttcagcgc gcgtcttcgt cacggatgag 5401 aatgacaacg cgcctgtctt cgcctcgccg tcacgcgtgc gcctcccaga ggaccagccg 5461 cctgggcccg cggccctgca cgtggtagcc cgggacccgg atctgggcga ggctgcacgc 5521 gtgtcctatc ggctggcatc tggcggggac ggccacttcc ggctgcactc aagcactgga 5581 gcgctgtccg tggtgcggcc gttggaccgc gaacaacgag ctgagcacgt actgacagtg 5641 gtggcctcag accacggctc cccgccgcgc tcggccacgc aggtcctgac cgtcagtgtc 5701 gctgacgtca acgacgaggc gcctactttc cagcagcagg agtacagcgt cctcttgcgt 5761 gagaacaacc ctcctggcac atctctgctc accctgcgag caaccgaccc cgacgtgggg 5821 gccaacgggc aagtgactta tggaggcgtc tctagcgaaa gcttttctct ggatcctgac 5881 actggtgttc tcacgactct tcgggccctg gatcgagagg aacaggagga gatcaacctg 5941 acagtgtatg cccaggacag gggctcacct cctcagttaa cgcatgtcac tgttcgagtg 6001 gctgtggagg atgagaatga ccatgcacca acctttggga gtgcccatct ctctctggag 6061 gtgcctgagg gccaggaccc ccagaccctt accatgcttc gggcctctga tccagatgtg 6121 ggagccaatg ggcagttgca gtaccgcatc ctagatgggg acccatcagg agcctttgtc 6181 ctagaccttg cttctggaga gtttggcacc atgcggccac tagacagaga agtggagcca 6241 gctttccagc tgaggataga ggcccgggat ggaggccagc cagctctcag tgccacgctg 6301 cttttgacag tgacagtgct ggatgccaat gaccatgctc cagcctttcc tgtgcctgcc 6361 tactcggtgg aggtgccgga ggatgtgcct gcagggaccc tgctgctgca gctacaggct 6421 catgaccctg atgctggagc taatggccat gtgacctact acctgggcgc cggtacagca 6481 ggagccttcc tgctggagcc cagctctgga gaactgcgca cagctgcagc cttggacaga 6541 gaacagtgtc ccagctacac cttttctgtg agtgcagtgg atggtgcagc tgctgggccc 6601 ctaagcacca cagtgtctgt caccatcatg gtgcgcgatg tcaatgacca tgcacccacc 6661 ttccccacca gtcctctgcg cctacgtctg ccccgcccag gccccagctt cagtacccca 6721 accctggctc tggccacact gagagctgaa gatcgtgatg ctggtgccaa tgcttccatt 6781 ctgtaccggc tggcaggcac accacctcct ggcactactg tggactctta cactggtgaa 6841 atccgcgtgg cccgctctcc tgtagctcta ggcccccgag atcgtgtcct cttcattgtg 6901 gccactgatc ttggccgtcc agctcgctct gccactggtg tgatcattgt tggactgcag 6961 ggggaagctg agcgtggacc ccgctttccc cgggctagca gtgaggctac gattcgtgag 7021 aatgcgcccc cagggactcc tattgtctcc cccagggccg tccatgcagg aggcacaaat 7081 ggacccatca cctacagcat tctcagtggg aatgagaaag ggacattctc catccagcct 7141 agtacaggtg ccatcacagt tcgctcagca gaggggctag acttcgaggt gagtccacgg 7201 ctgcgactgg tgctgcaggc agagagtgga ggagcctttg ccttcactgt gctgaccctg 7261 accctgcaag atgccaacga caatgctccc cgtttcctgc ggccccatta tgtggccttc 7321 cttcctgagt cccggccctt ggaggggccc ctgctgcagg tggaggcgga tgacctggat 7381 caaggctctg gaggacagat ttcctacagt ctggctgcat cccagccggc acgtggattg 7441 ttccacgtag acccaaccac aggcactatc actaccacag ccatcctgga ccgtgagatc 7501 tgggctgaaa cacggttggt gctgatggcc acagacagag ggagcccagc cctggtgggc 7561 tcagctacct tgacggtgat ggtcatcgac accaatgaca atcgccccac catcccccaa 7621 ccctgggagc tccgagtgtc agaagatgcg ttattgggct cagagattgc acaggtaaca 7681 gggaatgatg tggactcagg acccgtgctg tggtatgtgc taagcccatc tgggccccag 7741 gatcccttca gtgttggccg ctatggaggc cgtgtctccc tcacggggcc cctggacttt 7801 gagcagtgtg accgctacca gctgcagctg ctggcacatg atgggcctca tgagggccgt 7861 gccaacctca cagtgcttgt ggaggatgtc aatgacaatg cacctgcctt ctcacagagc 7921 ctctaccagg taatgctgct tgagcacaca cccccaggca gtgccattct ctccgtctct 7981 gccactgatc gggactcagg tgccaacggt cacatttcct accacctggc ttcccctgcc 8041 gatggcttca gtgttgaccc caacaatggg accctgttca caatagtggg aacagtggcc 8101 ttgggccatg acgggtcagg agcagtggat gtggtgctgg aagcacgaga ccacggggct 8161 ccaggccggg cagcacgagc cacagtgcac gtgcagctgc aggaccagaa cgaccacgcc 8221 ccgagcttca cattgtcaca ctaccgtgtg gctgtgactg aagacctgcc ccctggctcc 8281 actctgctca ccctggaggc tacagatgct gatggaagcc gcagccatgc cgctgtggac 8341 tacagcatca tcagtggcaa ctggggccga gtcttccagc tggaacccag gctggctgag 8401 gctggggaga gtgctggacc aggcccccgg gcactgggct gcctggtgtt gcttgaacct 8461 ctagactttg aaagcctgac acagtacaat ctaacagtgg ctgcagctga ccgtgggcag 8521 ccaccccaaa gctcagtcgt gccagtcact gtcactgtac tagatgtcaa tgacaaccca 8581 cctgtcttta cccgagcatc ctaccgtgtg acagtacctg aggacacacc tgttggagct 8641 gagctgctgc atgtagaggc ctctgacgct gaccctggcc ctcatggcct cgtgcgtttc 8701 actgtcagct caggcgaccc atcagggctc tttgagctgg atgagagctc aggcaccttg 8761 cgactggccc atgccctgga ctgtgagacc caggctcgac atcagcttgt agtacaggct 8821 gctgaccctg ctggtgcaca ctttgctttg gcaccagtga caattgaggt ccaggatgtg 8881 aatgatcatg gcccagcctt cccactgaac ttactcagca ccagcgtggc cgagaatcag 8941 cctccaggca ctctcgtgac cactctgcat gcaatcgacg gggatgctgg ggcttttggg 9001 aggctccgtt acagcctgtt ggaggctggg ccaggacctg agggccgtga ggcatttgca 9061 ctgaacagct caacagggga gttgcgtgcg cgagtgccct ttgactatga gcacacagaa 9121 agcttccggc tgctggtggg tgctgctgat gctgggaatc tctcagcctc tgtcaccgtg 9181 tcggtgctag tgactggaga ggatgagtat gaccctgtat ttctggcacc agctttccac 9241 ttccaagtgc ccgaaggtgc ccggcgtggc cacagcttgg gtcacgtgca ggccacagat 9301 gaggatgggg gtgccgatgg cctggttctg tattcccttg ccacctcttc cccctatttt 9361 ggtattaacc agactacagg agccctgtac ctgcgggtgg acagtcgggc accaggcagc 9421 ggaacagcca cctctggggg tgggggccgg acccggcggg aagcaccacg ggagctgagg 9481 ctggaggtga tagcacgggg gcctctgcct ggttcccgga gtgccacagt gcctgtgacc 9541 gtggatatca cccacaccgc actgggcctg gcacctgacc tcaacctgct attagtaggg 9601 gccgtggcag cctccttggg agttgtggtg gtgcttgcac tggcagccct ggtcctagga 9661 cttgttcggg cccgtagccg caaggctgag gcagcccctg gcccaatgtc acaggcagca 9721 cccctagcca gtgactcact gcagaaactg ggccgggagc cacctagtcc accaccctct 9781 gagcacctct atcaccagac tcttcccagc tatggtgggc caggagctgg aggaccctac 9841 ccccgtggtg gctccttgga cccttcacat tcaagtggcc gaggatcagc agaggctgca 9901 gaggatgatg agatccgcat gatcaatgag ttcccccgtg tggccagtgt ggcctcctct 9961 ctggctgccc gtggccctga ctcaggcatc cagcaggatg cagatggtct gagtgacaca 10021 tcctgcgaac cacctgcccc tgacacctgg tataagggcc gaaaggcagg gctgctgctg 10081 ccaggtgcag gagccactct ctacagagag gaggggcccc cagccactgc cacagccttc 10141 ctggggggct gtggcctgag ccctgcaccc actggggact atggcttccc agcagatggc 10201 aagccatgtg tggcaggtgc gctgacagcc attgtggccg gcgaggagga gctccgtggc 10261 agctataact gggactacct gctgagctgg tgccctcagt tccaaccact ggccagtgtc 10321 ttcacagaga tcgctcggct caaggatgaa gctcggccat gtcccccagc tccccgtatc 10381 gacccaccac ccctcatcac tgccgtggcc cacccaggag ccaagtctgt gccccccaag 10441 ccagcaaaca cagctgcagc ccgggccatc ttcccaccag cttctcaccg ctcccccatc 10501 agccatgaag gctccctgtc ctcagctgcc atgtccccca gcttctcacc ctctctgtct 10561 cctctggctg ctcgctcacc cgttgtctca ccatttgggg tggcccaggg tccctcagcc 10621 tcagcactca gcgcagagtc tggcctggag ccacctgatg acacggagct gcacatcacc 10681 ggtggctctg gcatggtgag caagggcgag gaggataaca tggccatcat caaggagttc 10741 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc 10801 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt 10861 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc 10921 tacgtgaagc accccgccga cattcccgac tacttgaagc tgtccttccc cgagggcttc 10981 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc 11041 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc 11101 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac 11161 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc 11221 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc 11281 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg 11341 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag 11401 tcttgactcg agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct 11461 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc 11521 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt 11581 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat 11641 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg 11701 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg 11761 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc 11821 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc 11881 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt 11941 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag 12001 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg 12061 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag 12121 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg 12181 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag 12241 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc 12301 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc 12361 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg 12421 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag 12481 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtga 12541 tgaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca 12601 gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg 12661 taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc 12721 gttatgttta tcggcacttt gcatcggccg cgctcccgat tccggaagtg cttgacattg 12781 gggaattcag cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc 12841 aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg 12901 cgatcgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa 12961 tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc 13021 actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc 13081 tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct 13141 ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga 13201 tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt 13261 gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc 13321 ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg 13381 gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag 13441 ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct 13501 gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg 13561 aatagcacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg 13621 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt 13681 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca 13741 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac 13801 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat 13861 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac 13921 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa 13981 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat 14041 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc 14101 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg 14161 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag 14221 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc 14281 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag 14341 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga 14401 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc 14461 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg 14521 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt 14581 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca 14641 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca 14701 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag 14761 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca 14821 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg 14881 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa 14941 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta 15001 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag 15061 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga 15121 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac 15181 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc 15241 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta 15301 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac 15361 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat 15421 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa 15481 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg 15541 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag 15601 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc 15661 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct 15721 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat 15781 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg 15841 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc 15901 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta 15961 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg 16021 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga 16081 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg 16141 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct 16201 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgtt //
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