CAPTURE2_pLVX-EF1a-dCas9-CBio-IRES-zsGreen1

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联系方式:I47-825O-882O

相关技术服务:质粒构建    基因合成    质粒大提

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载体基本信息

质粒名称: CAPTURE2_pLVX-EF1a-dCas9-CBio-IRES-zsGreen1 
出品公司: 爱迪基因
目录编号: 138418
存储实验室: Jian Xu

载体骨架基本信息
载体名称: pLVX-EF1a-zsGreen1
载体出品公司: Takara Bio
载体抗性: 氨苄青霉素
空载体大小: --
完整质粒大小: --
载体修饰: --
载体类型: 慢病毒载体
筛选标记: zsGreen1
高拷贝/低拷贝: --
生长条件: 37 ℃

插入基因信息
插入基因名称: dCas9
物种来源: Streptococcus pyogenes
插入大小: 4401 bp
基因突变: --
GenBank编号 --
启动子 EF1a
标签/融合蛋白: BioTAP-tag(插接件上的C端子)
克隆方法: 限制性内切酶
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
材料和方法部分: CAPTURE2_pLVX-EF1a-dCas9-CBio-IRES-zsGreen1 was a gift from Jian Xu (Addgene plasmid # 138418 ; http://n2t.net/addgene:138418 ; RRID:Addgene_138418)
参考文献部分: Multiplexed capture of spatial configuration and temporal dynamics of locus-specific 3D chromatin by biotinylated dCas9. Liu X, Chen Y, Zhang Y, Liu Y, Liu N, Botten GA, Cao H, Orkin SH, Zhang MQ, Xu J. Genome Biol. 2020 Mar 5;21(1):59. doi: 10.1186/s13059-020-01973-w. 10.1186/s13059-020-01973-w PubMed 32138752

载体图谱质粒图谱和多克隆位点信息

CAPTURE2_pLVX-EF1a-dCas9-CBio-IRES-zsGreen1质粒图谱

载体简介

载体序列

LOCUS       CAPTURE2_pLVX-EF1a-dCas9-CBio-IRES-zsGreen1               13286 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  CAPTURE2.0 vector containing dCas9-CBio, IRES and zsGreen1.
ACCESSION   .
VERSION     .
KEYWORDS    CAPTURE2_pLVX-EF1a-dCas9-CBio-IRES-zsGreen1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13286)
  AUTHORS   Liu X, Chen Y, Zhang Y, Liu Y, Liu N, Botten GA, Cao H, Orkin SH, 
            Zhang MQ, Xu J
  TITLE     Multiplexed capture of spatial configuration and temporal dynamics 
            of locus-specific 3D chromatin by biotinylated dCas9.
  JOURNAL   Genome Biol. 2020 Mar 5;21(1):59. doi: 10.1186/s13059-020-01973-w.
  PUBMED    32138752
REFERENCE   2  (bases 1 to 13286)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..13286
                     /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    2028..2143
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     promoter        2338..3519
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2568..3510
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3467..3487
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      3558..3567
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3573..3593
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3594..7694
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7704..7724
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    7972..8544
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    7995..8546
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(8139..8156)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             8546..9241
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein"
                     /label=ZsGreen1
                     /note="mammalian codon-optimized"
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
     misc_feature    9255..9843
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9308..9328)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9726..9737)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10050..10683
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10811..10827)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10811..10827)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10824..10846)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10835..10851
                     /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(10859..10889)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10904..10925
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11042..11059)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11213..11801)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11293..11312)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(11972..12832)
                     /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     12595..12614
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12833..12937)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    12985..13119
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13022..13041)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     13076..13095
                     /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 tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgatgagtaa ttcatacaaa aggactcgcc cctgccttgg
     2221 ggaatcccag ggaccgtcgt taaactccca ctaacgtaga acccagagat cgctgcgttc
     2281 ccgccccctc acccgcccgc tctcgtcatc actgaggtgg agaagagcat gcgtgaggct
     2341 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag
     2401 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg
     2461 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag
     2521 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt
     2581 gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt
     2641 ccacgcccct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
     2701 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
     2761 cttgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
     2821 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
     2881 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
     2941 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
     3001 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
     3061 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     3121 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     3181 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     3241 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     3301 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     3361 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     3421 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     3481 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gatctatttc cggtgaattc
     3541 ctcgagacta gttctaggcc accatggcta gccccaaaaa gaagaggaaa gtggacaaga
     3601 agtattctat cggactggcc atcgggacta atagcgtcgg gtgggccgtg atcactgacg
     3661 agtacaaggt gccctctaag aagttcaagg tgctcgggaa caccgaccgg cattccatca
     3721 agaaaaatct gatcggagct ctcctctttg attcagggga gaccgctgaa gcaacccgcc
     3781 tcaagcggac tgctagacgg cggtacacca ggaggaagaa ccggatttgt taccttcaag
     3841 agatattctc caacgaaatg gcaaaggtcg acgacagctt cttccatagg ctggaagaat
     3901 cattcctcgt ggaagaggat aagaagcatg aacggcatcc catcttcggt aatatcgtcg
     3961 acgaggtggc ctatcacgag aaatacccaa ccatctacca tcttcgcaaa aagctggtgg
     4021 actcaaccga caaggcagac ctccggctta tctacctggc cctggcccac atgatcaagt
     4081 tcagaggcca cttcctgatc gagggcgacc tcaatcctga caatagcgat gtggataaac
     4141 tgttcatcca gctggtgcag acttacaacc agctctttga agagaacccc atcaatgcaa
     4201 gcggagtcga tgccaaggcc attctgtcag cccggctgtc aaagagccgc agacttgaga
     4261 atcttatcgc tcagctgccg ggtgaaaaga aaaatggact gttcgggaac ctgattgctc
     4321 tttcacttgg gctgactccc aatttcaagt ctaatttcga cctggcagag gatgccaagc
     4381 tgcaactgtc caaggacacc tatgatgacg atctcgacaa cctcctggcc cagatcggtg
     4441 accaatacgc cgaccttttc cttgctgcta agaatctttc tgacgccatc ctgctgtctg
     4501 acattctccg cgtgaacact gaaatcacca aggcccctct ttcagcttca atgattaagc
     4561 ggtatgatga gcaccaccag gacctgaccc tgcttaaggc actcgtccgg cagcagcttc
     4621 cggagaagta caaggaaatc ttctttgacc agtcaaagaa tggatacgcc ggctacatcg
     4681 acggaggtgc ctcccaagag gaattttata agtttatcaa acctatcctt gagaagatgg
     4741 acggcaccga agagctcctc gtgaaactga atcgggagga tctgctgcgg aagcagcgca
     4801 ctttcgacaa tgggagcatt ccccaccaga tccatcttgg ggagcttcac gccatccttc
     4861 ggcgccaaga ggacttctac ccctttctta aggacaacag ggagaagatt gagaaaattc
     4921 tcactttccg catcccctac tacgtgggac ccctcgccag aggaaatagc cggtttgctt
     4981 ggatgaccag aaagtcagaa gaaactatca ctccctggaa cttcgaagag gtggtggaca
     5041 agggagccag cgctcagtca ttcatcgaac ggatgactaa cttcgataag aacctcccca
     5101 atgagaaggt cctgccgaaa cattccctgc tctacgagta ctttaccgtg tacaacgagc
     5161 tgaccaaggt gaaatatgtc accgaaggga tgaggaagcc cgcattcctg tcaggcgaac
     5221 aaaagaaggc aattgtggac cttctgttca agaccaatag aaaggtgacc gtgaagcagc
     5281 tgaaggagga ctatttcaag aaaattgaat gcttcgactc tgtggagatt agcggggtcg
     5341 aagatcggtt caacgcaagc ctgggtacct accatgatct gcttaagatc atcaaggaca
     5401 aggattttct ggacaatgag gagaacgagg acatccttga ggacattgtc ctgactctca
     5461 ctctgttcga ggaccgggaa atgatcgagg agaggcttaa gacctacgcc catctgttcg
     5521 acgataaagt gatgaagcaa cttaaacgga gaagatatac cggatgggga cgccttagcc
     5581 gcaaactcat caacggaatc cgggacaaac agagcggaaa gaccattctt gatttcctta
     5641 agagcgacgg attcgctaat cgcaacttca tgcaacttat ccatgatgat tccctgacct
     5701 ttaaggagga catccagaag gcccaagtgt ctggacaagg tgactcactg cacgagcata
     5761 tcgcaaatct ggctggttca cccgctatta agaagggtat tctccagacc gtgaaagtcg
     5821 tggacgagct ggtcaaggtg atgggtcgcc ataaaccaga gaacattgtc atcgagatgg
     5881 ccagggaaaa ccagactacc cagaagggac agaagaacag cagggagcgg atgaaaagaa
     5941 ttgaggaagg gattaaggag ctcgggtcac agatccttaa agagcacccg gtggaaaaca
     6001 cccagcttca gaatgagaag ctctatctgt actaccttca aaatggacgc gatatgtatg
     6061 tggaccaaga gcttgatatc aacaggctct cagactacga cgtggacgcc atcgtccctc
     6121 agagcttcct caaagacgac tcaattgaca ataaggtgct gactcgctca gacaagaacc
     6181 ggggaaagtc agataacgtg ccctcagagg aagtcgtgaa aaagatgaag aactattggc
     6241 gccagcttct gaacgcaaag ctgatcactc agcggaagtt cgacaatctc actaaggctg
     6301 agaggggcgg actgagcgaa ctggacaaag caggattcat taaacggcaa cttgtggaga
     6361 ctcggcagat tactaaacat gtcgcccaaa tccttgactc acgcatgaat accaagtacg
     6421 acgaaaacga caaacttatc cgcgaggtga aggtgattac cctgaagtcc aagctggtca
     6481 gcgatttcag aaaggacttt caattctaca aagtgcggga gatcaataac tatcatcatg
     6541 ctcatgacgc atatctgaat gccgtggtgg gaaccgccct gatcaagaag tacccaaagc
     6601 tggaaagcga gttcgtgtac ggagactaca aggtctacga cgtgcgcaag atgattgcca
     6661 aatctgagca ggagatcgga aaggccaccg caaagtactt cttctacagc aacatcatga
     6721 atttcttcaa gaccgaaatc acccttgcaa acggtgagat ccggaagagg ccgctcatcg
     6781 agactaatgg ggagactggc gaaatcgtgt gggacaaggg cagagatttc gctaccgtgc
     6841 gcaaagtgct ttctatgcct caagtgaaca tcgtgaagaa aaccgaggtg caaaccggag
     6901 gcttttctaa ggaatcaatc ctccccaagc gcaactccga caagctcatt gcaaggaaga
     6961 aggattggga ccctaagaag tacggcggat tcgattcacc aactgtggct tattctgtcc
     7021 tggtcgtggc taaggtggaa aaaggaaagt ctaagaagct caagagcgtg aaggaactgc
     7081 tgggtatcac cattatggag cgcagctcct tcgagaagaa cccaattgac tttctcgaag
     7141 ccaaaggtta caaggaagtc aagaaggacc ttatcatcaa gctcccaaag tatagcctgt
     7201 tcgaactgga gaatgggcgg aagcggatgc tcgcctccgc tggcgaactt cagaagggta
     7261 atgagctggc tctcccctcc aagtacgtga atttcctcta ccttgcaagc cattacgaga
     7321 agctgaaggg gagccccgag gacaacgagc aaaagcaact gtttgtggag cagcataagc
     7381 attatctgga cgagatcatt gagcagattt ccgagttttc taaacgcgtc attctcgctg
     7441 atgccaacct cgataaagtc cttagcgcat acaataagca cagagacaaa ccaattcggg
     7501 agcaggctga gaatatcatc cacctgttca ccctcaccaa tcttggtgcc cctgccgcat
     7561 tcaagtactt cgacaccacc atcgaccgga aacgctatac ctccaccaaa gaagtgctgg
     7621 acgccaccct catccaccag agcatcaccg gactttacga aactcggatt gacctctcac
     7681 agctcggagg ggatgaggga gctcccaaga aaaagcgcaa ggtaggtagt tccgccggaa
     7741 aggccggtga aggtgaaatc cctgcccctc ttgctggtac cgtttctaag atactggtaa
     7801 aagaaggtga cactgttaaa gctggtcaaa cagttctggt gctggaggct atgaaaatgg
     7861 agacagaaat taacgctcct actgacggaa aagttgaaaa ggtgttagtt aaggaaagag
     7921 atgctgttca aggtggtcaa ggtctaatca agatcggcgt ttaagatccc gcccctctcc
     7981 ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt
     8041 ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg
     8101 ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg
     8161 tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc
     8221 tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca
     8281 aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag
     8341 ttggatagtt gtggaaagag tcaaatggct cacctcaagc gtattcaaca aggggctgaa
     8401 ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt
     8461 tacatgtgtt tagtcgaggt taaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt
     8521 ttcctttgaa aaacacgatg ataatatggc ccagtccaag cacggcctga ccaaggagat
     8581 gaccatgaag taccgcatgg agggctgcgt ggacggccac aagttcgtga tcaccggcga
     8641 gggcatcggc taccccttca agggcaagca ggccatcaac ctgtgcgtgg tggagggcgg
     8701 ccccttgccc ttcgccgagg acatcttgtc cgccgccttc atgtacggca accgcgtgtt
     8761 caccgagtac ccccaggaca tcgtcgacta cttcaagaac tcctgccccg ccggctacac
     8821 ctgggaccgc tccttcctgt tcgaggacgg cgccgtgtgc atctgcaacg ccgacatcac
     8881 cgtgagcgtg gaggagaact gcatgtacca cgagtccaag ttctacggcg tgaacttccc
     8941 cgccgacggc cccgtgatga agaagatgac cgacaactgg gagccctcct gcgagaagat
     9001 catccccgtg cccaagcagg gcatcttgaa gggcgacgtg agcatgtacc tgctgctgaa
     9061 ggacggtggc cgcttgcgct gccagttcga caccgtgtac aaggccaagt ccgtgccccg
     9121 caagatgccc gactggcact tcatccagca caagctgacc cgcgaggacc gcagcgacgc
     9181 caagaaccag aagtggcacc tgaccgagca cgccatcgcc tccggctccg ccttgccctg
     9241 aacgcgtctg gaacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     9301 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     9361 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     9421 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     9481 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     9541 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     9601 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct
     9661 ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     9721 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     9781 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     9841 cgcctggaat taattctgca gtcgagacct agaaaaacat ggagcaatca caagtagcaa
     9901 tacagcagct accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg
     9961 ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct
    10021 tagccacttt ttaaaagaaa agaggggact ggaagggcta attcactccc aacgaagaca
    10081 agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt agcagaacta
    10141 cacaccaggg ccaggggtca gatatccact gacctttgga tggtgctaca agctagtacc
    10201 agttgagcca gataaggtag aagaggccaa taaaggagag aacaccagct tgttacaccc
    10261 tgtgagcctg catgggatgg atgacccgga gagagaagtg ttagagtgga ggtttgacag
    10321 ccgcctagca tttcatcacg tggcccgaga gctgcatccg gagtacttca agaactgctg
    10381 atatcgagct tgctacaagg gactttccgc tggggacttt ccagggaggc gtggcctggg
    10441 cgggactggg gagtggcgag ccctcagatc ctgcatataa gcagctgctt tttgcctgta
    10501 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
    10561 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
    10621 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
    10681 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat
    10741 cagagagtga gaggccttga cattgctagc gtttaccgtc gacctctagc tagagcttgg
    10801 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
    10861 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
    10921 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
    10981 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
    11041 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
    11101 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
    11161 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
    11221 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
    11281 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
    11341 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
    11401 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
    11461 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
    11521 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
    11581 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
    11641 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
    11701 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
    11761 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
    11821 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
    11881 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
    11941 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
    12001 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
    12061 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
    12121 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
    12181 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
    12241 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
    12301 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
    12361 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
    12421 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
    12481 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
    12541 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
    12601 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
    12661 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
    12721 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
    12781 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
    12841 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
    12901 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
    12961 ctgacgtcga cggatcggga gatcaacttg tttattgcag cttataatgg ttacaaataa
    13021 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
    13081 ttgtccaaac tcatcaatgt atcttatcat gtctggatca actggataac tcaagctaac
    13141 caaaatcatc ccaaacttcc caccccatac cctattacca ctgccaatta cctgtggttt
    13201 catttactct aaacctgtga ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt
    13261 taaatatgta ctacaaactt agtagt
//

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