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产品名称：pCDNA-dCas9-VP64
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产品展商：XYbscience
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简单介绍

pCDNA-dCas9-VP64质粒是CRISPR/Cas9系列质粒，复制子是f1 ori、ori和SV40 ori，质粒大小是9812bp，带有Ampicillin氨苄青霉素抗性，pCDNA-dCas9-VP64可转化进Stbl3中培养，培养条件是LB培养基，37℃。

产品描述

pCDNA-dCas9-VP64

产品信息
基本信息
质粒简介
质粒图谱
质粒序列

产品信息

产品货号	产品名称	产品规格	优惠价
XY0118	pCDNA-dCas9-VP64	5μg	¥请询价

使用说明

信裕质粒平台的各批次质粒菌株发货前均经过严格的多重验证，如存在质量问题，请在收到产品的三个月内通知我司。收到质粒后请短暂离心，加入20μl ddH2O溶解质粒，取2μl转化至对应感受态中，挑取单克隆重新提取质粒后使用。如需获取其他详细信息请登录我司官网查询。

基本信息

启动子:	CMV promoter
复制子:	F1 ori，ori，SV40 ori
终止子:	bGH poly(A) signal
质粒分类:	哺乳细胞质粒；哺乳编辑质粒；哺乳Cas9质粒
质粒大小:	9812bp
质粒标签:	N-3×Flag,N-NLS,C-VP64,C-HA
原核抗性:	氨苄青霉素Amp（100μg/ml）
筛选标记:	新霉素Neo/G418
克隆菌株:	Stbl3等大肠杆菌
培养条件:	37℃，LB，有氧
表达宿主:	293T等哺乳细胞
培养条件:	37℃，5%CO2
诱导方式:	无需诱导
5'测序引物:	CMV-F(CGCAAATGGGCGGTAGGCGTG)
3'测序引物:	BGH-R（TAGAAGGCACAGTCGAGG）

质粒简介

pCDNA-dCas9-VP64质粒是CRISPR/Cas9系列质粒，复制子是f1 ori、ori和SV40 ori，质粒大小是9812bp，带有Ampicillin氨苄青霉素抗性，可转化进Stbl3中培养，培养条件是LB培养基，37℃。

As evidence that the dCas9-VP64–gRNA system can activate gene expression in other cell types, we transfected expression plasmids for dCas9-VP64 and the four gRNAs targeting Ascl1 into mouse embryonic fibroblasts. Because the gRNA target sites are conserved in the human ASCL1 and mouse Ascl1 promoters, we also observed activation of Ascl1 expression 4 d after transfection in MEFs treated with plasmids encoding dCas9-VP64 and the four gRNAs。gene activation by dCas9-VP64 and combinations of gRNAs can be very specific, this is not necessarily a surrogate for DNA-binding, and additional studies are necessary to define the role of the gRNA target sequence in determining genome-wide Cas9 activity in mammalian cells. The RNA-seq results for both IL1RN and HBG1-HBG2 showed moderate downregulation of the IL32 gene (false discovery rate < 0.03) in the samples treated with dCas9-VP64 and gRNA expression plasmids compared to control samples treated with only an empty expression plasmid. Because both the IL1RNtargetedandHBG1-HBG2–targeted samples were similarlyaffected, it is unlikely that this is the result of off-target dCas9-VP64 activity related to the identity of the target sequences. Weconfirmed these results by qRT-PCR for IL32, which showed thatthis downregulation is a general response to dCas9-VP64, evenin the absence of gRNAs.

质粒图谱

质粒序列

LOCUS       Exported                9812 bp ds-DNA     circular SYN 29-JUN-2017
DEFINITION synthetic circular DNA
KEYWORDS    pcDNA-dCas9-VP64
SOURCE      synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE   1 (bases 1 to 9812)
TITLE     Direct Submission
JOURNAL   Exported Thursday, June 29, 2017 from MiaoLingPlasmid
            http://www.miaolingbio.com
FEATURES             Location/Qualifiers
     source          1..9812
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        235..614
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        863..881
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             907..972
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /note="3xFLAG"
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             979..999
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T
                     antigen"
                     /note="SV40 NLS"
                     /translation="PKKKRKV"
     CDS             1009..5112
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9
                     endonuclease from the Streptococcus pyogenes Type II
                     CRISPR/Cas system"
                     /note="dCas9"
                     /note="RNA-guided DNA-binding protein that lacks
                     endonuclease activity due to the D10 mutation in the
                     RuvC-like domain and the H840A mutation in the HNH domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5137..5157
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T
                     antigen"
                     /note="SV40 NLS"
                     /translation="PKKKRKV"
     CDS             5167..5316
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /note="VP64"
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             5323..5349
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /note="HA"
                     /translation="YPYDVPDYA"
     polyA_signal    5412..5636
                     /note="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5682..6110
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6124..6453
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      6304..6439
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     CDS             6520..7314
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="NeoR/KanR"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    7488..7609
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7658..7674)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    7682..7698
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /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(7706..7736)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    7751..7772
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(8060..8645)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(8816..9676)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(9677..9781)
                     /gene="bla"
                     /note="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gccatggact acaaagacca tgacggtgat tataaagatc atgacatcga ttacaaggat
      961 gacgatgaca agatggcccc caagaagaag aggaaggtgg gccgcggaat ggataagaaa
     1021 tactcaatag gcttagctat cggcacaaat agcgtcggat gggcggtgat cactgatgaa
     1081 tataaggttc cgtctaaaaa gttcaaggtt ctgggaaata cagaccgcca cagtatcaaa
     1141 aaaaatctta taggggctct tttatttgac agtggagaga cagcggaagc gactcgtctc
     1201 aaacggacag ctcgtagaag gtatacacgt cggaagaatc gtatttgtta tctacaggag
     1261 attttttcaa atgagatggc gaaagtagat gatagtttct ttcatcgact tgaagagtct
     1321 tttttggtgg aagaagacaa gaagcatgaa cgtcatccta tttttggaaa tatagtagat
     1381 gaagttgctt atcatgagaa atatccaact atctatcatc tgcgaaaaaa attggtagat
     1441 tctactgata aagcggattt gcgcttaatc tatttggcct tagcgcatat gattaagttt
     1501 cgtggtcatt ttttgattga gggagattta aatcctgata atagtgatgt ggacaaacta
     1561 tttatccagt tggtacaaac ctacaatcaa ttatttgaag aaaaccctat taacgcaagt
     1621 ggagtagatg ctaaagcgat tctttctgca cgattgagta aatcaagacg attagaaaat
     1681 ctcattgctc agctccccgg tgagaagaaa aatggcttat ttgggaatct cattgctttg
     1741 tcattgggtt tgacccctaa ttttaaatca aattttgatt tggcagaaga tgctaaatta
     1801 cagctttcaa aagatactta cgatgatgat ttagataatt tattggcgca aattggagat
     1861 caatatgctg atttgttttt ggcagctaag aatttatcag atgctatttt actttcagat
     1921 atcctaagag taaatactga aataactaag gctcccctat cagcttcaat gattaaacgc
     1981 tacgatgaac atcatcaaga cttgactctt ttaaaagctt tagttcgaca acaacttcca
    2041 gaaaagtata aagaaatctt ttttgatcaa tcaaaaaacg gatatgcagg ttatattgat
     2101 gggggagcta gccaagaaga attttataaa tttatcaaac caattttaga aaaaatggat
     2161 ggtactgagg aattattggt gaaactaaat cgtgaagatt tgctgcgcaa gcaacggacc
     2221 tttgacaacg gctctattcc ccatcaaatt cacttgggtg agctgcatgc tattttgaga
     2281 agacaagaag acttttatcc atttttaaaa gacaatcgtg agaagattga aaaaatcttg
     2341 acttttcgaa ttccttatta tgttggtcca ttggcgcgtg gcaatagtcg ttttgcatgg
     2401 atgactcgga agtctgaaga aacaattacc ccatggaatt ttgaagaagt tgtcgataaa
     2461 ggtgcttcag ctcaatcatt tattgaacgc atgacaaact ttgataaaaa tcttccaaat
     2521 gaaaaagtac taccaaaaca tagtttgctt tatgagtatt ttacggttta taacgaattg
     2581 acaaaggtca aatatgttac tgaaggaatg cgaaaaccag catttctttc aggtgaacag
     2641 aagaaagcca ttgttgattt actcttcaaa acaaatcgaa aagtaaccgt taagcaatta
     2701 aaagaagatt atttcaaaaa aatagaatgt tttgatagtg ttgaaatttc aggagttgaa
     2761 gatagattta atgcttcatt aggtacctac catgatttgc taaaaattat taaagataaa
     2821 gattttttgg ataatgaaga aaatgaagat atcttagagg atattgtttt aacattgacc
     2881 ttatttgaag atagggagat gattgaggaa agacttaaaa catatgctca cctctttgat
     2941 gataaggtga tgaaacagct taaacgtcgc cgttatactg gttggggacg tttgtctcga
     3001 aaattgatta atggtattag ggataagcaa tctggcaaaa caatattaga ttttttgaaa
     3061 tcagatggtt ttgccaatcg caattttatg cagctgatcc atgatgatag tttgacattt
     3121 aaagaagaca ttcaaaaagc acaagtgtct ggacaaggcg atagtttaca tgaacatatt
     3181 gcaaatttag ctggtagccc tgctattaaa aaaggtattt tacagactgt aaaagttgtt
     3241 gatgaattgg tcaaagtaat ggggcggcat aagccagaaa atatcgttat tgaaatggca
     3301 cgtgaaaatc agacaactca aaagggccag aaaaattcgc gagagcgtat gaaacgaatc
     3361 gaagaaggta tcaaagaatt aggaagtcag attcttaaag agcatcctgt tgaaaatact
     3421 caattgcaaa atgaaaagct ctatctctat tatctccaaa atggaagaga catgtatgtg
     3481 gaccaagaat tagatattaa tcgtttaagt gattatgatg tcgatgccat tgttccacaa
     3541 agtttcctta aagacgattc aatagacaat aaggtcttaa cgcgttctga taaaaatcgt
     3601 ggtaaatcgg ataacgttcc aagtgaagaa gtagtcaaaa agatgaaaaa ctattggaga
     3661 caacttctaa acgccaagtt aatcactcaa cgtaagtttg ataatttaac gaaagctgaa
     3721 cgtggaggtt tgagtgaact tgataaagct ggttttatca aacgccaatt ggttgaaact
     3781 cgccaaatca ctaagcatgt ggcacaaatt ttggatagtc gcatgaatac taaatacgat
     3841 gaaaatgata aacttattcg agaggttaaa gtgattacct taaaatctaa attagtttct
     3901 gacttccgaa aagatttcca attctataaa gtacgtgaga ttaacaatta ccatcatgcc
     3961 catgatgcgt atctaaatgc cgtcgttgga actgctttga ttaagaaata tccaaaactt
     4021 gaatcggagt ttgtctatgg tgattataaa gtttatgatg ttcgtaaaat gattgctaag
     4081 tctgagcaag aaataggcaa agcaaccgca aaatatttct tttactctaa tatcatgaac
     4141 ttcttcaaaa cagaaattac acttgcaaat ggagagattc gcaaacgccc tctaatcgaa
     4201 actaatgggg aaactggaga aattgtctgg gataaagggc gagattttgc cacagtgcgc
     4261 aaagtattgt ccatgcccca agtcaatatt gtcaagaaaa cagaagtaca gacaggcgga
     4321 ttctccaagg agtcaatttt accaaaaaga aattcggaca agcttattgc tcgtaaaaaa
     4381 gactgggatc caaaaaaata tggtggtttt gatagtccaa cggtagctta ttcagtccta
     4441 gtggttgcta aggtggaaaa agggaaatcg aagaagttaa aatccgttaa agagttacta
     4501 gggatcacaa ttatggaaag aagttccttt gaaaaaaatc cgattgactt tttagaagct
     4561 aaaggatata aggaagttaa aaaagactta atcattaaac tacctaaata tagtcttttt
     4621 gagttagaaa acggtcgtaa acggatgctg gctagtgccg gagaattaca aaaaggaaat
     4681 gagctggctc tgccaagcaa atatgtgaat tttttatatt tagctagtca ttatgaaaag
     4741 ttgaagggta gtccagaaga taacgaacaa aaacaattgt ttgtggagca gcataagcat
     4801 tatttagatg agattattga gcaaatcagt gaattttcta agcgtgttat tttagcagat
     4861 gccaatttag ataaagttct tagtgcatat aacaaacata gagacaaacc aatacgtgaa
     4921 caagcagaaa atattattca tttatttacg ttgacgaatc ttggagctcc cgctgctttt
     4981 aaatattttg atacaacaat tgatcgtaaa cgatatacgt ctacaaaaga agttttagat
     5041 gccactctta tccatcaatc catcactggt ctttatgaaa cacgcattga tttgagtcag
     5101 ctaggaggtg acccaattgc cggatccaag gctagcccga aaaagaaacg caaagttggg
     5161 cgcgccgacg cgctggacga tttcgatctc gacatgctgg gttctgatgc cctcgatgac
     5221 tttgacctgg atatgttggg aagcgacgca ttggatgact ttgatctgga catgctcggc
     5281 tccgatgctc tggacgattt cgatctcgat atgttaatta actacccgta cgacgttccg
     5341 gactacgctt cttgaatcgg taggaattcg cggccgtcta gacttaagtt taaaccgctg
     5401 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     5461 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     5521 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     5581 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     5641 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     5701 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     5761 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     5821 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
    5881 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     5941 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     6001 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     6061 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     6121 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     6181 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     6241 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     6301 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     6361 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     6421 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     6481 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     6541 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     6601 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     6661 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     6721 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     6781 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     6841 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     6901 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     6961 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     7021 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     7081 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     7141 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     7201 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     7261 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     7321 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     7381 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     7441 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     7501 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     7561 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     7621 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     7681 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     7741 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     7801 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     7861 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     7921 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     7981 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     8041 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     8101 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     8161 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     8221 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     8281 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     8341 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     8401 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     8461 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     8521 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     8581 ccaccgctgg tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     8641 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     8701 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     8761 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     8821 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     8881 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     8941 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     9001 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     9061 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     9121 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     9181 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     9241 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     9301 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     9361 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     9421 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     9481 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     9541 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     9601 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     9661 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     9721 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     9781 gcacatttcc ccgaaaagtg ccacctgacg tc
//

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