质粒名称: | pHAGE-IRES-puro-NLS-dPspCas13b-EGFP-NLS-3xFlag |
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出品公司: | Addgene |
目录编号: | 132397 |
存储实验室: | Ling-Ling Chen |
载体骨架基本信息 |
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载体名称: | pHAGE-EF1a-IRES-puro |
载体出品公司: | -- |
空载体大小: | -- |
完整质粒大小: | -- |
载体修饰: | -- |
载体类型: | 哺乳动物表达,慢病毒 |
筛选标记: | -- |
载体抗性: | 氨苄青霉素 |
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: | dPspCas13b |
物种来源: | Prevotella sp. P5-125 |
插入大小: | -- |
基因突变: | H133A; H1058A |
GenBank编号 | -- |
启动子: | -- |
标签/融合蛋白: | flag(插入件上的N端子) |
克隆方法: | Ligation Independent 克隆 |
5’克隆位点: | -- |
3’克隆位点: | -- |
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: | pHAGE-IRES-puro-NLS-dPspCas13b-EGFP-NLS-3xFlag was a gift from Ling-Ling Chen (Addgene plasmid # 132397 ; http://n2t.net/addgene:132397 ; RRID:Addgene_132397) |
参考文献部分: | dCas13 imaging. Liang-Zhong Yang, Yang Wang, Si-Qi Li, Huang Wu, Gordon G. Carmichael, Ling-Ling Chen. Molecular Cell 10.1016/j.molcel.2019.10.024 |
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 pHAGE-IRES-puro-NLS-dPspCas13b-EGFP-NLS-3xFlag 12449 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION overexpression in human cells. ACCESSION . VERSION . KEYWORDS pHAGE-IRES-puro-NLS-dPspCas13b-EGFP-NLS-3xFlag SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12449) AUTHORS Liang-Zhong Yang, Yang Wang, Si-Qi Li, Huang Wu, Gordon G. Carmichael, Ling-Ling Chen TITLE dCas13 imaging JOURNAL Molecular Cell REFERENCE 2 (bases 1 to 12449) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..12449 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 1..634 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 681..806 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1303..1536 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1721..1765 /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 2058..2175 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2223..3401 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 2454..3392 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 3349..3369 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" CDS 3431..3451 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" regulatory 6746..6755 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 6752..7468 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 6752..7468 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(6797..6818) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(7058..7077) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 7405..7426 /label=EGFP-C /note="EGFP, forward primer" CDS 7496..7543 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" CDS 7562..7627 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" misc_feature 7634..8201 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(7795..7812) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 8022..8041 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 8221..8820 /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(8221..8240) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 8717..8737 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 8839..9427 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(8892..8912) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(9310..9321) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 9502..9735 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" primer_bind complement(9854..9872) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 9940..10044 /gene="bla" /label=AmpR promoter CDS 10045..10905 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(10263..10282) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 11076..11664 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 11565..11584 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 11818..11835 /label=L4440 /note="L4440 vector, forward primer" primer_bind 11927..11946 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" protein_bind 12075..12096 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 12111..12141 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 12149..12165 /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 12154..12176 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 12173..12189 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 12173..12189 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 12239..12258 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" ORIGIN 1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca 61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag 661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg 841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg 901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct 961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat 1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca 1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc 1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag 1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc 1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg 1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc 1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc 1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct 1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa 1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca 1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt 1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt 1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta 1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt 1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct 1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga 1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg 2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa 2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca 2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtcgt 2221 gaggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg 2281 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa 2341 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg 2401 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg 2461 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa 2521 ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt 2581 gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct 2641 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg 2701 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct 2761 ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg 2821 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc 2881 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc 2941 ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac 3001 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga 3061 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc 3121 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg 3181 attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga 3241 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt 3301 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga 3361 cagtggttca aagttttttt cttccatttc aggtgtcgtg aagcggccgc accggtctgc 3421 agctagcatg cccaaaaaga agaggaaagt gaacatcccc gctctggtgg aaaaccagaa 3481 gaagtacttt ggcacctaca gcgtgatggc catgctgaac gctcagaccg tgctggacca 3541 catccagaag gtggccgata ttgagggcga gcagaacgag aacaacgaga atctgtggtt 3601 tcaccccgtg atgagccacc tgtacaacgc caagaacggc tacgacaagc agcccgagaa 3661 aaccatgttc atcatcgagc ggctgcagag ctacttccca ttcctgaaga tcatggccga 3721 gaaccagaga gagtacagca acggcaagta caagcagaac cgcgtggaag tgaacagcaa 3781 cgacatcttc gaggtgctga agcgcgcctt cggcgtgctg aagatgtaca gggacctgac 3841 caacgcctac aagacctacg aggaaaagct gaacgacggc tgcgagttcc tgaccagcac 3901 agagcaacct ctgagcggca tgatcaacaa ctactacaca gtggccctgc ggaacatgaa 3961 cgagagatac ggctacaaga cagaggacct ggccttcatc caggacaagc ggttcaagtt 4021 cgtgaaggac gcctacggca agaaaaagtc ccaagtgaat accggattct tcctgagcct 4081 gcaggactac aacggcgaca cacagaagaa gctgcacctg agcggagtgg gaatcgccct 4141 gctgatctgc ctgttcctgg acaagcagta catcaacatc tttctgagca ggctgcccat 4201 cttctccagc tacaatgccc agagcgagga acggcggatc atcatcagat ccttcggcat 4261 caacagcatc aagctgccca aggaccggat ccacagcgag aagtccaaca agagcgtggc 4321 catggatatg ctcaacgaag tgaagcggtg ccccgacgag ctgttcacaa cactgtctgc 4381 cgagaagcag tcccggttca gaatcatcag cgacgaccac aatgaagtgc tgatgaagcg 4441 gagcagcgac agattcgtgc ctctgctgct gcagtatatc gattacggca agctgttcga 4501 ccacatcagg ttccacgtga acatgggcaa gctgagatac ctgctgaagg ccgacaagac 4561 ctgcatcgac ggccagacca gagtcagagt gatcgagcag cccctgaacg gcttcggcag 4621 actggaagag gccgagacaa tgcggaagca agagaacggc accttcggca acagcggcat 4681 ccggatcaga gacttcgaga acatgaagcg ggacgacgcc aatcctgcca actatcccta 4741 catcgtggac acctacacac actacatcct ggaaaacaac aaggtcgaga tgtttatcaa 4801 cgacaaagag gacagcgccc cactgctgcc cgtgatcgag gatgatagat acgtggtcaa 4861 gacaatcccc agctgccgga tgagcaccct ggaaattcca gccatggcct tccacatgtt 4921 tctgttcggc agcaagaaaa ccgagaagct gatcgtggac gtgcacaacc ggtacaagag 4981 actgttccag gccatgcaga aagaagaagt gaccgccgag aatatcgcca gcttcggaat 5041 cgccgagagc gacctgcctc agaagatcct ggatctgatc agcggcaatg cccacggcaa 5101 ggatgtggac gccttcatca gactgaccgt ggacgacatg ctgaccgaca ccgagcggag 5161 aatcaagaga ttcaaggacg accggaagtc cattcggagc gccgacaaca agatgggaaa 5221 gagaggcttc aagcagatct ccacaggcaa gctggccgac ttcctggcca aggacatcgt 5281 gctgtttcag cccagcgtga acgatggcga gaacaagatc accggcctga actaccggat 5341 catgcagagc gccattgccg tgtacgatag cggcgacgat tacgaggcca agcagcagtt 5401 caagctgatg ttcgagaagg cccggctgat cggcaagggc acaacagagc ctcatccatt 5461 tctgtacaag gtgttcgccc gcagcatccc cgccaatgcc gtcgagttct acgagcgcta 5521 cctgatcgag cggaagttct acctgaccgg cctgtccaac gagatcaaga aaggcaacag 5581 agtggatgtg cccttcatcc ggcgggacca gaacaagtgg aaaacacccg ccatgaagac 5641 cctgggcaga atctacagcg aggatctgcc cgtggaactg cccagacaga tgttcgacaa 5701 tgagatcaag tcccacctga agtccctgcc acagatggaa ggcatcgact tcaacaatgc 5761 caacgtgacc tatctgatcg ccgagtacat gaagagagtg ctggacgacg acttccagac 5821 cttctaccag tggaaccgca actaccggta catggacatg cttaagggcg agtacgacag 5881 aaagggctcc ctgcagcact gcttcaccag cgtggaagag agagaaggcc tctggaaaga 5941 gcgggcctcc agaacagagc ggtacagaaa gcaggccagc aacaagatcc gcagcaaccg 6001 gcagatgaga aacgccagca gcgaagagat cgagacaatc ctggataagc ggctgagcaa 6061 cagccggaac gagtaccaga aaagcgagaa agtgatccgg cgctacagag tgcaggatgc 6121 cctgctgttt ctgctggcca aaaagaccct gaccgaactg gccgatttcg acggcgagag 6181 gttcaaactg aaagaaatca tgcccgacgc cgagaaggga atcctgagcg agatcatgcc 6241 catgagcttc accttcgaga aaggcggcaa gaagtacacc atcaccagcg agggcatgaa 6301 gctgaagaac tacggcgact tctttgtgct ggctagcgac aagaggatcg gcaacctgct 6361 ggaactcgtg ggcagcgaca tcgtgtccaa agaggatatc atggaagagt tcaacaaata 6421 cgaccagtgc aggcccgaga tcagctccat cgtgttcaac ctggaaaagt gggccttcga 6481 cacatacccc gagctgtctg ccagagtgga ccgggaagag aaggtggact tcaagagcat 6541 cctgaaaatc ctgctgaaca acaagaacat caacaaagag cagagcgaca tcctgcggaa 6601 gatccggaac gccttcgatg ccaacaatta ccccgacaaa ggcgtggtgg aaatcaaggc 6661 cctgcctgag atcgccatga gcatcaagaa ggcctttggg gagtacgcca tcatgaagga 6721 gggagctcta gaggcgctac cggtcgccac catggtgagc aagggcgagg agctgttcac 6781 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt 6841 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac 6901 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca 6961 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc 7021 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg 7081 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga 7141 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa 7201 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca 7261 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg 7321 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa 7381 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat 7441 cactctcggc atggacgagc tgtacaagtc gaattctaag ccaggattcc atatgaagcg 7501 acctgccgcc acaaagaagg ctggacaggc taagaagaag aaacaacgcg tgcagctaag 7561 cgactacaaa gaccatgacg gtgattataa agatcatgac atcgattaca aggatgacga 7621 tgacaagtaa ggatccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag 7681 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga 7741 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg 7801 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt 7861 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca 7921 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc 7981 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat 8041 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc 8101 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga 8161 accacgggga cgtggttttc ctttgaaaaa cacgatgata aggatccacc ggagcttacc 8221 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta 8281 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac 8341 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac 8401 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag 8461 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt 8521 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag 8581 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc 8641 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg 8701 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc 8761 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga 8821 cgcccatcga tagatcctaa tcaacctctg gattacaaaa tttgtgaaag attgactggt 8881 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat 8941 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg 9001 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt 9061 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact 9121 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc 9181 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg 9241 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc 9301 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg 9361 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc 9421 tccccgcctg agatccttta agaccaatga cttacaaggc agctgtagat cttagccact 9481 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc 9541 tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct 9601 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt 9661 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt 9721 ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa 9781 agaaatgaat atcagagagt gagaggcccg ggttaattaa ggaaagggct agatcattct 9841 tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg 9901 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta 9961 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt 10021 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc 10081 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa 10141 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt 10201 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt 10261 ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc 10321 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg 10381 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg 10441 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac 10501 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca 10561 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta 10621 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat 10681 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa 10741 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag 10801 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat 10861 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt 10921 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg 10981 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga 11041 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta 11101 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa 11161 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact 11221 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca 11281 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt 11341 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg 11401 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag 11461 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta 11521 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat 11581 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg 11641 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc 11701 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac 11761 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc 11821 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt 11881 tggccgattc attaatgcag caagctcatg gctgactaat tttttttatt tatgcagagg 11941 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc 12001 taggcttttg caaaaagctc cccgtggcac gacaggtttc ccgactggaa agcgggcagt 12061 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt 12121 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac 12181 agctatgaca tgattacgaa tttcacaaat aaagcatttt tttcactgca ttctagttgt 12241 ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcaactggat aactcaagct 12301 aaccaaaatc atcccaaact tcccacccca taccctatta ccactgccaa ttacctgtgg 12361 tttcatttac tctaaacctg tgattcctct gaattatttt cattttaaag aaattgtatt 12421 tgttaaatat gtactacaaa cttagtagt //
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