质粒名称: | p5xUAS-Flag-NPM1wt |
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出品公司: | addgene |
目录编号: | 131822 |
存储实验室: | Mark Minden |
载体骨架基本信息 |
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载体名称: | 5xUAS 4-OHTinducible system |
完整质粒大小: | 10194bp |
载体类型: | Lentiviral |
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: | NPM1 |
物种来源: | H. sapiens (human) |
标签/融合蛋白: | Flag (N terminal on insert) |
文献引用方法 |
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材料和方法部分: | p5xUAS-Flag-NPM1wt was a gift from Mark Minden (Addgene plasmid # 131822 ; http://n2t.net/addgene:131822 ; RRID:Addgene_131822) |
参考文献部分: | NPM1c impedes CTCF functions through cytoplasmic mislocalization in acute myeloid leukemia. Wang AJ, Han Y, Jia N, Chen P, Minden MD. Leukemia. 2020 May;34(5):1278-1290. doi: 10.1038/s41375-019-0681-8. Epub 2019 Dec 12. 10.1038/s41375-019-0681-8 PubMed 31831844 |
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载体序列
LOCUS p5xUAS-Flag-NPM1wt 10194 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION 4-hydroxytamoxifen inducible vector expressing Flag-NPM1wildtype (human) in mammalian cells through transduction. ACCESSION . VERSION . KEYWORDS p5xUAS-Flag-NPM1wt SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10194) TITLE NPM1c Impedes CTCF Functions through Cytoplasmic Mislocalization in Acute Myeloid Leukemia JOURNAL Unpublished REFERENCE 2 (bases 1 to 10194) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..10194 /organism="synthetic DNA construct" /mol_type="other DNA" polyA_signal 501..622 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(538..557) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 592..611 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(671..687) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(671..687) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(684..706) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 695..711 /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(719..749) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 764..785 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(902..919) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1073..1661) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1153..1172) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1832..2692) /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 2455..2474 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2693..2797) /gene="bla" /label=AmpR promoter primer_bind complement(2872..2891) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 3063..3442 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 3444..3642 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 3597..3617 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 3660..3840 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 3887..4012 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 4505..4738 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 4923..4967 /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 5265..5382 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" protein_bind 5447..5541 /label=5X UAS /bound_moiety="GAL4" /note="five tandem copies of the ""ScaI site"" 17-mer CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) that efficiently binds yeast Gal4 (Webster et al., 1988; Pfeiffer et al., 2010)" protein_bind 5466..5560 /label=5X UAS /bound_moiety="GAL4" /note="five tandem copies of the ""ScaI site"" 17-mer CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) that efficiently binds yeast Gal4 (Webster et al., 1988; Pfeiffer et al., 2010)" primer_bind 5606..5622 /label=pBMN 5' /note="MMLV sequence, for inserts in pBMN retroviral vector" regulatory 5666..5675 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 5675..5698 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" promoter 6647..6976 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 6827..6962 /label=SV40 ori /note="SV40 origin of replication" CDS 6986..7585 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(6986..7005) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 7482..7502 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 7711..8299 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(7764..7784) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(8182..8193) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(8302..8318) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(8303..8319) /label=pBluescriptKS /note="For pBluescript vector" LTR 8824..9004 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(9030..9047) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 9036..9260 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 9306..9734 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(9393..9412) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 9603..9624 /label=F1ori-F /note="F1 origin, forward primer" promoter 9748..10077 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 9928..10063 /label=SV40 ori /note="SV40 origin of replication" promoter 10125..10172 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 10191..371 /codon_start=1 /gene="Sh ble from Streptoalloteichus hindustanus" /product="antibiotic-binding protein" /label=BleoR /note="confers resistance to bleomycin, phleomycin, and Zeocin(TM)" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" ORIGIN 1 ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt 61 tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg 121 tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca 181 ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg 241 tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt 301 gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg 361 agcaggactg acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg 421 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc 481 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca 541 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt 601 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg 661 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca 721 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca 781 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc 841 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt 901 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact 961 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag 1021 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata 1081 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc 1141 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg 1201 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc 1261 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg 1321 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc 1381 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga 1441 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg 1501 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa 1561 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg 1621 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt 1681 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat 1741 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct 1801 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta 1861 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa 1921 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac 1981 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa 2041 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag 2101 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg 2161 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag 2221 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg 2281 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc 2341 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat 2401 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata 2461 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa 2521 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca 2581 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc 2641 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc 2701 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg 2761 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac 2821 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg 2881 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga 2941 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa 3001 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg 3061 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag 3121 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc 3181 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg 3241 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca 3301 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc 3361 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt 3421 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata 3481 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt 3541 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca 3601 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agcgcgtttt gcctgtactg 3661 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac 3721 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt 3781 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca 3841 gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag 3901 gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc 3961 aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa 4021 gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa 4081 atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc 4141 tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct 4201 tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt 4261 gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca 4321 aaacaaaagt aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag 4381 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat 4441 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg 4501 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt 4561 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca 4621 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca 4681 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg 4741 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt 4801 ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag 4861 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag 4921 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta 4981 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag 5041 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac 5101 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag 5161 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatcggcac 5221 tgcgtgcgcc aattctgcag acaaatggca gtattcatcc acaattttaa aagaaaaggg 5281 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa 5341 actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac 5401 agcagagatc cagtttggtt aattaatagc ttgcatgctt gcaggtcgga gtactgtcct 5461 ccgagcggag tactgtcctc cgagcggagt actgtcctcc gagcggagta ctgtcctccg 5521 agcggagtac tgtcctccga gcggagtact gtcctccgag cggagactct agcgggtata 5581 taatgagatc cagttatcca tcgcagcttg gatacacgcc gcccacgtga aggctgccga 5641 ccccgggggt ggaccatccg gatccgccac catggattac aaggatgacg acgataagac 5701 cggtatggaa gattcgatgg acatggacat gagccccctg aggccccaga actatctttt 5761 cggttgtgaa ctaaaggccg acaaagatta tcactttaag gtggataatg atgaaaatga 5821 gcaccagtta tctttaagaa cggtcagttt aggggctggt gcaaaggatg agttgcacat 5881 tgttgaagca gaggcaatga attacgaagg cagtccaatt aaagtaacac tggcaacttt 5941 gaaaatgtct gtacagccaa cggtttccct tgggggcttt gaaataacac caccagtggt 6001 cttaaggttg aagtgtggtt cagggccagt gcatattagt ggacagcact tagtagctgt 6061 ggaggaagat gcagagtcag aagatgaaga ggaggaggat gtgaaactct taagtatatc 6121 tggaaagcgg tctgcccctg gaggtggtag caaggttcca cagaaaaaag taaaacttgc 6181 tgctgatgaa gatgatgacg atgatgatga agaggatgat gatgaagatg atgatgatga 6241 tgattttgat gatgaggaag ctgaagaaaa agcgccagtg aagaaatcta tacgagatac 6301 tccagccaaa aatgcacaaa agtcaaatca gaatggaaaa gactcaaaac catcatcaac 6361 accaagatca aaaggacaag aatccttcaa gaaacaggaa aaaactccta aaacaccaaa 6421 aggacctagt tctgtagaag acattaaagc aaaaatgcaa gcaagtatag aaaaaggtgg 6481 ttctcttccc aaagtggaag ccaaattcat caattatgtg aagaattgct tccggatgac 6541 tgaccaagag gctattcaag atctctggca gtggaggaag tctctttaag aaaatagacc 6601 ggttagtcta gagatccgcg gccgcatcga ttcgaccctg tggaatgtgt gtcagttagg 6661 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta 6721 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat 6781 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac 6841 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga 6901 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg 6961 cctaggcttt tgcaaaaagc ttaccatgac cgagtacaag cccacggtgc gcctcgccac 7021 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc 7081 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact 7141 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc 7201 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg 7261 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct 7321 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc 7381 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga 7441 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga 7501 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg 7561 catgacccgc aagcccggtg cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag 7621 gagcgcacga ccccatgcat cgattagtcc aatttgttaa agacaggata acctcgaggg 7681 cgcgccgaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa 7741 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta 7801 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa 7861 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg 7921 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc 7981 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc 8041 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg 8101 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg 8161 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg 8221 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc 8281 ctttgggccg cctccccgca tcgataccgt cgacctcgag acctagaaaa acatggagca 8341 atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga agcacaagag 8401 gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat gacttacaag 8461 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac 8521 tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct 8581 gattggcaga actacacacc agggccaggg atcagatatc cactgacctt tggatggtgc 8641 tacaagctag taccagttga gcaagagaag gtagaagaag ccaatgaagg agagaacacc 8701 cgcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga agtattagag 8761 tggaggtttg acagccgcct agcatttcat cacatggccc gagagctgca tccggactgt 8821 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac 8881 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg 8941 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct 9001 agcagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc 9061 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct 9121 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg 9181 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg 9241 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta 9301 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt 9361 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct 9421 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg 9481 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag 9541 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa 9601 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga 9661 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa 9721 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc 9781 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga 9841 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 9901 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat 9961 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc 10021 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag 10081 ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat 10141 cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc atgg //