pHAGE-TO-dCas9

价格:1000元

联系方式:I47-825O-882O

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质粒基本信息

质粒名称: pHAGE-TO-dCas9 
出品公司: 爱迪基因
目录编号: 75381
存储实验室: Thoru Pederson

载体骨架基本信息
载体名称: pHAGE-DEST
载体抗性: 氨苄霉素
空载体大小: 6000 bp
完整质粒大小: 10829 bp
载体类型: 哺乳细胞表达,Lentiviral
高拷贝/低拷贝: 高拷贝
生长条件: 37 ℃

插入基因信息
插入基因名称: Sp dCas9
物种来源: Synthetic
插入大小: 4101 bp
启动子 CMV-TO
标签/融合蛋白: no
克隆方法: 限制性内切酶
5' 克隆位点: EcoRI(未破坏)
3' 克隆位点: XbaI(未破坏)
5' 测序引物 cgaaggaatagaagaagaaggtgga
3' 测序引物 CCAAAGGGAGATCCGACTCG

文献引用方法
材料和方法部分: pHAGE-TO-dCas9 was a gift from Thoru Pederson (爱迪基因 plasmid # 75381)
参考文献部分: Multiplexed labeling of genomic loci with dCas9 and engineered sgRNAs using CRISPRainbow. Ma H, Tu LC, Naseri A, Huisman M, Zhang S, Grunwald D, Pederson T. Nat Biotechnol. 2016 Apr 18. doi: 10.1038/nbt.3526. 10.1038/nbt.3526 PubMed 27088723

质粒图谱信息

pHAGE-TO-dCas9质粒图谱

质粒简介

质粒序列

LOCUS       pHAGE-TO-dCas9               10835 bp ds-DNA     circular SYN 10-JAN-2018
DEFINITION  dCas9.
ACCESSION   .
VERSION     .
KEYWORDS    pHAGE-TO-dCas9
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10835)
  AUTHORS   Ma H, Tu LC, Naseri A, Huisman M, Zhang S, Grunwald D, Pederson T
  TITLE     Multiplexed labeling of genomic loci with dCas9 and engineered 
            sgRNAs using CRISPRainbow.
  JOURNAL   Nat Biotechnol. 2016 Apr 18. doi: 10.1038/nbt.3526.
  PUBMED    27088723
COMMENT     This file is created by Vector NTI
            http://www.biofeng.com/
COMMENT     ORIGDB|GenBank
COMMENT     VNTAUTHORNAME|biofeng.com|
COMMENT     VNTNAME|pHAGE-TO-dCas9|
FEATURES             Location/Qualifiers
     source          1..10835
                     /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."
     misc_feature    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2299..2602
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2603..2806
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2757..2777
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2808..2826
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2829..2847
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2857..2881
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2974..2983
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2986..3006
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3025..7125
                     /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             7144..7164
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    7225..7813
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7278..7298)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7696..7707)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7888..8121
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8240..8258)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8326..8430
                     /gene="bla"
                     /label=AmpR promoter
     CDS             8431..9291
                     /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(8649..8668)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9462..10050
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9951..9970
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10204..10221
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     10313..10332
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     protein_bind    10461..10482
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        10497..10527
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10535..10551
                     /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     10540..10562
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10559..10575
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10559..10575
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10625..10644
                     /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 ttgacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagcgat
     2221 gctagctgat gcgggcccta gttattaata gtaatcaatt acggggtcat tagttcatag
     2281 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     2341 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     2401 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     2461 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     2521 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     2581 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
     2641 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     2701 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     2761 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctcc ctatcagtga
     2821 tagagatctc cctatcagtg atagagatcg tcgacgagct cgtttagtga accgtcagat
     2881 cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg accgatccag
     2941 cctaccggtt aaggtgaacg cgtgcaggct ggcgccacca tggcccctaa aaagaaacgc
     3001 aaggtcgaat tcggaagcgg aagcgacaag aagtactcca ttgggctcgc tatcggtacc
     3061 aacagcgtcg gctgggccgt cattacggac gagtacaagg tgccgagcaa aaaattcaaa
     3121 gttctgggca ataccgatcg ccacagcata aagaagaacc tcattggagc cctcctgttc
     3181 gactccgggg agacggccga agccacgcgg ctcaaaagaa cagcacggcg cagatatacc
     3241 cgcagaaaga atcggatctg ctacctgcag gagatcttta gtaatgagat ggctaaggtg
     3301 gatgactctt tcttccatag gctggaggag tcctttttgg tggaggagga taaaaagcac
     3361 gagcgccacc caatctttgg caatatcgtg gacgaggtgg cgtaccatga aaagtaccca
     3421 accatatatc atctgaggaa gaagctggta gacagtactg ataaggctga cttgcggttg
     3481 atctatctcg cgctggcgca catgatcaaa tttcggggac acttcctcat cgagggggac
     3541 ctgaacccag acaacagcga tgtcgacaaa ctctttatcc aactggttca gacttacaat
     3601 cagcttttcg aggagaaccc gatcaacgca tccggcgttg acgccaaagc aatcctgagc
     3661 gctaggctgt ccaaatcccg gcggctcgaa aacctcatcg cacagctccc tggggagaag
     3721 aagaacggcc tgtttggtaa tcttatcgcc ctgtcactcg ggctgacccc caactttaaa
     3781 tctaacttcg acctggccga agatgccaag ctgcaactga gcaaagacac ctacgatgat
     3841 gatctcgaca atctgctggc ccagatcggc gaccagtacg cagacctttt tttggcggca
     3901 aagaacctgt cagacgccat tctgctgagt gatattctgc gagtgaacac ggagatcacc
     3961 aaagctccgc tgagcgctag tatgatcaag cgctatgatg agcaccacca agacttgact
     4021 ttgctgaagg cccttgtcag acagcaactg cctgagaagt acaaggaaat tttcttcgat
     4081 cagtctaaaa atggctacgc cggatacatt gacggcggag caagccagga ggaattttac
     4141 aaatttatta agcccatctt ggaaaaaatg gacggcaccg aggagctgct ggtaaagctg
     4201 aacagagaag atctgttgcg caaacagcgc actttcgaca atggaagcat cccccaccag
     4261 attcacctgg gcgaactgca cgctatcctc aggcggcaag aggatttcta cccctttttg
     4321 aaagataaca gggaaaagat tgagaaaatc ctcacatttc ggatacccta ctatgtaggc
     4381 cccctcgctc ggggaaattc cagattcgcg tggatgactc gcaaatcaga agagaccatc
     4441 actccctgga acttcgagga agtcgtggat aagggggcct ctgcccagtc cttcatcgaa
     4501 aggatgacta actttgataa aaatctgcct aacgaaaagg tgcttcctaa acactctctg
     4561 ctgtacgagt acttcacagt ttataacgag ctcaccaagg tcaaatacgt cacagaaggg
     4621 atgagaaagc cagcattcct gtctggagag cagaagaaag ctatcgtgga cctcctcttc
     4681 aagacgaacc ggaaagttac cgtgaaacag ctcaaagaag actatttcaa aaagattgaa
     4741 tgtttcgact ctgttgaaat cagcggagtg gaggatcgct tcaacgcatc cctgggaacg
     4801 tatcacgatc tcctgaaaat cattaaagac aaggacttcc tggacaatga ggagaacgag
     4861 gacattcttg aggacattgt cctcaccctt acgttgtttg aagataggga gatgattgaa
     4921 gaacgcttga aaacttacgc tcatctcttc gacgacaaag tcatgaaaca gctcaagaga
     4981 cgccgatata caggatgggg gcggctgtca agaaaactga tcaatggcat ccgagacaag
     5041 cagagtggaa agacaatcct ggattttctt aagtccgatg gatttgccaa ccggaacttc
     5101 atgcagttga tccatgatga ctctctcacc tttaaggagg acatccagaa agcacaagtt
     5161 tctggccagg gggacagtct tcacgagcac atcgctaatc ttgcaggtag cccagctatc
     5221 aaaaagggaa tactgcagac cgttaaggtc gtggatgaac tcgtcaaagt aatgggaagg
     5281 cataagcccg agaatatcgt tatcgagatg gcccgagaga accaaactac ccagaaggga
     5341 cagaagaaca gtagggaaag gatgaagagg attgaagagg gtataaaaga actggggtcc
     5401 caaatcctta aggaacaccc agttgaaaac acccagcttc agaatgagaa gctctacctg
     5461 tactacctgc agaacggcag ggacatgtac gtggatcagg aactggacat caaccggttg
     5521 tccgactacg acgtggatgc tatcgtgccc caaagctttc tcaaagatga ttctattgat
     5581 aataaagtgt tgacaagatc cgataaaaat agagggaaga gtgataacgt cccctcagaa
     5641 gaagttgtca agaaaatgaa aaattattgg cggcagctgc tgaacgccaa actgatcaca
     5701 caacggaagt tcgataatct gactaaggct gaacgaggtg gcctgtctga gttggataaa
     5761 gccggcttca tcaaaaggca gcttgttgag acacgccaga tcaccaagca cgtggcccaa
     5821 attctcgatt cacgcatgaa caccaagtac gatgaaaatg acaaactgat tcgagaggtg
     5881 aaagttatta ctctgaagtc taagctggtc tcagatttca gaaaggactt tcagttttat
     5941 aaggtgagag agatcaacaa ttaccaccat gcgcatgatg cctacctgaa tgcagtggta
     6001 ggcactgcac ttatcaaaaa atatcccaag ctggaatctg aatttgttta cggagactat
     6061 aaagtgtacg atgttaggaa aatgatcgca aagtctgagc aggaaatagg caaggccacc
     6121 gctaagtact tcttttacag caatattatg aattttttca agaccgagat tacactggcc
     6181 aatggagaga ttcggaagcg accacttatc gaaacaaacg gagaaacagg agaaatcgtg
     6241 tgggacaagg gtagggattt cgcgacagtc cgcaaggtcc tgtccatgcc gcaggtgaac
     6301 atcgttaaaa agaccgaagt acagaccgga ggcttctcca aggaaagtat cctcccgaaa
     6361 aggaacagcg acaagctgat cgcacgcaaa aaagattggg accccaagaa atacggcgga
     6421 ttcgattctc ctacagtcgc ttacagtgta ctggttgtgg ccaaagtgga gaaagggaag
     6481 tctaaaaaac tcaaaagcgt caaggaactg ctgggcatca caatcatgga gcgatccagc
     6541 ttcgagaaaa accccatcga ctttctcgaa gcgaaaggat ataaagaggt caaaaaagac
     6601 ctcatcatta agctgcccaa gtactctctc tttgagcttg aaaacggccg gaaacgaatg
     6661 ctcgctagtg cgggcgagct gcagaaaggt aacgagctgg cactgccctc taaatacgtt
     6721 aatttcttgt atctggccag ccactatgaa aagctcaaag ggtctcccga agataatgag
     6781 cagaagcagc tgttcgtgga acaacacaaa cactaccttg atgagatcat cgagcaaata
     6841 agcgagttct ccaaaagagt gatcctcgcc gacgctaacc tcgataaggt gctttctgct
     6901 tacaataagc acagggataa gcccatcagg gagcaggcag aaaacattat ccacttgttt
     6961 actctgacca acttgggcgc gcctgcagcc ttcaagtact tcgacaccac catagacaga
     7021 aagcggtaca cctctacaaa ggaggtcctg gacgccacac tgattcatca gtcaattacg
     7081 gggctctatg aaacaagaat cgacctctct cagctcggtg gagacggctc tactagtggc
     7141 tctcccaaga agaagaggaa ggtaggatcc ggatctggcg gccgctctgg catcgatggg
     7201 agcctcgagt agggtaatct agataatcaa cctctggatt acaaaatttg tgaaagattg
     7261 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     7321 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     7381 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     7441 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     7501 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     7561 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     7621 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     7681 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     7741 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     7801 gccgcctccc cgcctgagat cctttaagac caatgactta caaggcagct gtagatctta
     7861 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag
     7921 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     7981 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     8041 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     8101 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac
     8161 ttgcaaagaa atgaatatca gagagtgaga ggcccgggtt aattaaggaa agggctagat
     8221 cattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     8281 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     8341 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     8401 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     8461 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     8521 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     8581 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     8641 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     8701 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     8761 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     8821 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     8881 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     8941 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     9001 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     9061 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     9121 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     9181 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     9241 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     9301 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     9361 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     9421 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     9481 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     9541 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     9601 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     9661 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     9721 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     9781 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     9841 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     9901 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     9961 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
    10021 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
    10081 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
    10141 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
    10201 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
    10261 gcgcgttggc cgattcatta atgcagcaag ctcatggctg actaattttt tttatttatg
    10321 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
    10381 gaggcctagg cttttgcaaa aagctccccg tggcacgaca ggtttcccga ctggaaagcg
    10441 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
    10501 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
    10561 ggaaacagct atgacatgat tacgaatttc acaaataaag catttttttc actgcattct
    10621 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcaa ctggataact
    10681 caagctaacc aaaatcatcc caaacttccc accccatacc ctattaccac tgccaattac
    10741 ctgtggtttc atttactcta aacctgtgat tcctctgaat tattttcatt ttaaagaaat
    10801 tgtatttgtt aaatatgtac tacaaactta gtagt
//

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