pET-Cas9-NLS-6xHis

价格:1000元

联系方式:I47-825O-882O

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载体基本信息

出品公司: Addgene
载体名称: pET-Cas9-NLS-6xHis
质粒类型: CRISPR/Cas9载体;细菌基因敲除载体
高拷贝/低拷贝: 高拷贝
插入位点: 限制性内切酶,多克隆位点
启动子: lac/T7
载体大小: 9265 bp
5' 测序引物及序列: T7 forward:5’TAATACGACTCACTATAGGG 3’
3' 测序引物及序列: T7 Terminal:5’GCTAGTTATTGCTCAGCGG 3’
载体标签: NLS 、 His tag (C-ter)
载体抗性: Ampicillin
克隆菌株: Mach1
宿主细胞(系): 细菌,如大肠杆菌
备注:  
产品目录号: #62933
稳定性: 稳表达
组成型/诱导型: 诱导型
病毒/非病毒: 非病毒

载体质粒图谱和多克隆位点信息

pET-Cas9-NLS-6xHis 载体图谱



载体简介

载体序列

LOCUS       Exported                9265 bp ds-DNA     circular SYN 26-JUL-2016
DEFINITION  Bacterial vector for regulated expression of S. pyogenes Cas9 with a
            C-terminal nuclear localization signal and 6xHis tag.
ACCESSION   .
VERSION     .
KEYWORDS    pET-Cas9-NLS-6xHis
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9265)
  AUTHORS   Zuris JA, Thompson DB, Shu Y, Guilinger JP, Bessen JL, Hu JH, Maeder
            ML, Joung JK, Chen ZY, Liu DR.
  TITLE     Cationic lipid-mediated delivery of proteins enables efficient 
            protein-based genome editing in vitro and in vivo.
  JOURNAL   Nat. Biotechnol. 2015;33:73-80.
  PUBMED    25357182
REFERENCE   2  (bases 1 to 9265)
  AUTHORS   Liu Lab / Addgene #62933
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, July 26, 2016 from SnapGene Viewer 3.1.4
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9265
                     /organism="synthetic DNA construct"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
     protein_bind    3..27
                     /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)."
     RBS             42..64
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             71..4174
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /note="Cas9"
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             4175..4195
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /note="SV40 NLS"
                     /translation="PKKKRKV"
     CDS             4205..4222
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     terminator      4307..4354
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4391..4846
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(4961..5821)
                     /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(5822..5914)
                     /gene="bla"
                     /note="AmpR promoter"
     rep_origin      5995..6583
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7013..7204)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /note="rop"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(7776..8858)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /note="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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(8859..8936)
                     /gene="lacI"
                     /note="lacI promoter"
     promoter        9249..2
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
       61 gagatatacc atggataaga aatactcaat aggcttagat atcggcacaa atagcgtcgg
      121 atgggcggtg atcactgatg aatataaggt tccgtctaaa aagttcaagg ttctgggaaa
      181 tacagaccgc cacagtatca aaaaaaatct tataggggct cttttatttg acagtggaga
      241 gacagcggaa gcgactcgtc tcaaacggac agctcgtaga aggtatacac gtcggaagaa
      301 tcgtatttgt tatctacagg agattttttc aaatgagatg gcgaaagtag atgatagttt
      361 ctttcatcga cttgaagagt cttttttggt ggaagaagac aagaagcatg aacgtcatcc
      421 tatttttgga aatatagtag atgaagttgc ttatcatgag aaatatccaa ctatctatca
      481 tctgcgaaaa aaattggtag attctactga taaagcggat ttgcgcttaa tctatttggc
      541 cttagcgcat atgattaagt ttcgtggtca ttttttgatt gagggagatt taaatcctga
      601 taatagtgat gtggacaaac tatttatcca gttggtacaa acctacaatc aattatttga
      661 agaaaaccct attaacgcaa gtggagtaga tgctaaagcg attctttctg cacgattgag
      721 taaatcaaga cgattagaaa atctcattgc tcagctcccc ggtgagaaga aaaatggctt
      781 atttgggaat ctcattgctt tgtcattggg tttgacccct aattttaaat caaattttga
      841 tttggcagaa gatgctaaat tacagctttc aaaagatact tacgatgatg atttagataa
      901 tttattggcg caaattggag atcaatatgc tgatttgttt ttggcagcta agaatttatc
      961 agatgctatt ttactttcag atatcctaag agtaaatact gaaataacta aggctcccct
     1021 atcagcttca atgattaaac gctacgatga acatcatcaa gacttgactc ttttaaaagc
     1081 tttagttcga caacaacttc cagaaaagta taaagaaatc ttttttgatc aatcaaaaaa
     1141 cggatatgca ggttatattg atgggggagc tagccaagaa gaattttata aatttatcaa
     1201 accaatttta gaaaaaatgg atggtactga ggaattattg gtgaaactaa atcgtgaaga
     1261 tttgctgcgc aagcaacgga cctttgacaa cggctctatt ccccatcaaa ttcacttggg
     1321 tgagctgcat gctattttga gaagacaaga agacttttat ccatttttaa aagacaatcg
     1381 tgagaagatt gaaaaaatct tgacttttcg aattccttat tatgttggtc cattggcgcg
     1441 tggcaatagt cgttttgcat ggatgactcg gaagtctgaa gaaacaatta ccccatggaa
     1501 ttttgaagaa gttgtcgata aaggtgcttc agctcaatca tttattgaac gcatgacaaa
     1561 ctttgataaa aatcttccaa atgaaaaagt actaccaaaa catagtttgc tttatgagta
     1621 ttttacggtt tataacgaat tgacaaaggt caaatatgtt actgaaggaa tgcgaaaacc
     1681 agcatttctt tcaggtgaac agaagaaagc cattgttgat ttactcttca aaacaaatcg
     1741 aaaagtaacc gttaagcaat taaaagaaga ttatttcaaa aaaatagaat gttttgatag
     1801 tgttgaaatt tcaggagttg aagatagatt taatgcttca ttaggtacct accatgattt
     1861 gctaaaaatt attaaagata aagatttttt ggataatgaa gaaaatgaag atatcttaga
     1921 ggatattgtt ttaacattga ccttatttga agatagggag atgattgagg aaagacttaa
     1981 aacatatgct cacctctttg atgataaggt gatgaaacag cttaaacgtc gccgttatac
     2041 tggttgggga cgtttgtctc gaaaattgat taatggtatt agggataagc aatctggcaa
     2101 aacaatatta gattttttga aatcagatgg ttttgccaat cgcaatttta tgcagctgat
     2161 ccatgatgat agtttgacat ttaaagaaga cattcaaaaa gcacaagtgt ctggacaagg
     2221 cgatagttta catgaacata ttgcaaattt agctggtagc cctgctatta aaaaaggtat
     2281 tttacagact gtaaaagttg ttgatgaatt ggtcaaagta atggggcggc ataagccaga
     2341 aaatatcgtt attgaaatgg cacgtgaaaa tcagacaact caaaagggcc agaaaaattc
     2401 gcgagagcgt atgaaacgaa tcgaagaagg tatcaaagaa ttaggaagtc agattcttaa
     2461 agagcatcct gttgaaaata ctcaattgca aaatgaaaag ctctatctct attatctcca
     2521 aaatggaaga gacatgtatg tggaccaaga attagatatt aatcgtttaa gtgattatga
     2581 tgtcgatcac attgttccac aaagtttcct taaagacgat tcaatagaca ataaggtctt
     2641 aacgcgttct gataaaaatc gtggtaaatc ggataacgtt ccaagtgaag aagtagtcaa
     2701 aaagatgaaa aactattgga gacaacttct aaacgccaag ttaatcactc aacgtaagtt
     2761 tgataattta acgaaagctg aacgtggagg tttgagtgaa cttgataaag ctggttttat
     2821 caaacgccaa ttggttgaaa ctcgccaaat cactaagcat gtggcacaaa ttttggatag
     2881 tcgcatgaat actaaatacg atgaaaatga taaacttatt cgagaggtta aagtgattac
     2941 cttaaaatct aaattagttt ctgacttccg aaaagatttc caattctata aagtacgtga
     3001 gattaacaat taccatcatg cccatgatgc gtatctaaat gccgtcgttg gaactgcttt
     3061 gattaagaaa tatccaaaac ttgaatcgga gtttgtctat ggtgattata aagtttatga
     3121 tgttcgtaaa atgattgcta agtctgagca agaaataggc aaagcaaccg caaaatattt
     3181 cttttactct aatatcatga acttcttcaa aacagaaatt acacttgcaa atggagagat
     3241 tcgcaaacgc cctctaatcg aaactaatgg ggaaactgga gaaattgtct gggataaagg
     3301 gcgagatttt gccacagtgc gcaaagtatt gtccatgccc caagtcaata ttgtcaagaa
     3361 aacagaagta cagacaggcg gattctccaa ggagtcaatt ttaccaaaaa gaaattcgga
     3421 caagcttatt gctcgtaaaa aagactggga tccaaaaaaa tatggtggtt ttgatagtcc
     3481 aacggtagct tattcagtcc tagtggttgc taaggtggaa aaagggaaat cgaagaagtt
     3541 aaaatccgtt aaagagttac tagggatcac aattatggaa agaagttcct ttgaaaaaaa
     3601 tccgattgac tttttagaag ctaaaggata taaggaagtt aaaaaagact taatcattaa
     3661 actacctaaa tatagtcttt ttgagttaga aaacggtcgt aaacggatgc tggctagtgc
     3721 cggagaatta caaaaaggaa atgagctggc tctgccaagc aaatatgtga attttttata
     3781 tttagctagt cattatgaaa agttgaaggg tagtccagaa gataacgaac aaaaacaatt
     3841 gtttgtggag cagcataagc attatttaga tgagattatt gagcaaatca gtgaattttc
     3901 taagcgtgtt attttagcag atgccaattt agataaagtt cttagtgcat ataacaaaca
     3961 tagagacaaa ccaatacgtg aacaagcaga aaatattatt catttattta cgttgacgaa
     4021 tcttggagct cccgctgctt ttaaatattt tgatacaaca attgatcgta aacgatatac
     4081 gtctacaaaa gaagttttag atgccactct tatccatcaa tccatcactg gtctttatga
     4141 aacacgcatt gatttgagtc agctaggagg tgaccccaag aagaagagga aggtgatgga
     4201 taagcatcac caccaccatc actaatgata atttgaacgc cagcacatgg actcgtctac
     4261 tagcgcagct taattaacct aggctgctgc caccgctgag caataactag cataacccct
     4321 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatt
     4381 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     4441 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     4501 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     4561 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     4621 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     4681 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     4741 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     4801 caaaaattta acgcgaattt taacaaaata ttaacgttta caatttctgg cggcacgatg
     4861 gcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4921 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4981 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5041 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5101 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5161 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5221 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5281 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5341 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5401 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5461 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5521 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5581 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5641 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5701 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5761 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5821 tactcttcct ttttcaatca tgattgaagc atttatcagg gttattgtct catgagcgga
     5881 tacatatttg aatgtattta gaaaaataaa caaataggtc atgaccaaaa tcccttaacg
     5941 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     6001 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     6061 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6121 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     6181 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6241 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6301 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6361 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6421 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6481 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6541 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6601 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     6661 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     6721 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
     6781 ttttctcctt acgcatctgt gcggtatttc acaccgcata tatggtgcac tctcagtaca
     6841 atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg
     6901 tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     6961 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     7021 tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt
     7081 gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa
     7141 gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg
     7201 tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac
     7261 gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt
     7321 gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc
     7381 aatgccagcg cttcgttaat acagatgtag gtgttccaca gggtagccag cagcatcctg
     7441 cgatgcagat ccggaacata atggtgcagg gcgctgactt ccgcgtttcc agactttacg
     7501 aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc
     7561 agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaaccagta aggcaacccc
     7621 gccagcctag ccgggtcctc aacgacagga gcacgatcat gctagtcatg ccccgcgccc
     7681 accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct
     7741 aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     7801 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     7861 ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc
     7921 accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga
     7981 aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg
     8041 tatcccacta ccgagatgtc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt
     8101 gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc
     8161 agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct
     8221 atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc
     8281 gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga
     8341 tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc
     8401 tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg
     8461 gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga
     8521 ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg
     8581 ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc
     8641 agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt
     8701 gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt
     8761 ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg
     8821 gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc
     8881 tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg
     8941 atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag
     9001 gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc
     9061 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg
     9121 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
     9181 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatcg atctcgatcc
     9241 cgcgaaatta atacgactca ctata
//

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