pBbdCas9S_arr2

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pBbdCas9Sarr2载体基本信息

质粒名称: pBbdCas9S_arr2
出品公司: Addgene
目录编号: 149639
存储实验室: Christine Jacobs-Wagner

载体骨架基本信息
载体名称: pBbdCas9S
载体出品公司: CJW lab
空载体大小: --
完整质粒大小: --
载体修饰: dCas9, lacI, arr2
载体类型: 细菌表达,CRISPR
筛选标记: 利福平
载体抗性: 大观霉素
生长温度: 30 ℃
宿主菌: NEB 5-alpha F' Iq
拷贝数: 高拷贝

插入基因信息
插入基因名称: --
物种来源: --
插入大小: --
基因突变: --
GenBank编号 --
启动子: PflaB, PpQE30
标签/融合蛋白: --
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: pBbdCas9S_arr2 was a gift from Christine Jacobs-Wagner (Addgene plasmid # 149639 ; http://n2t.net/addgene:149639 ; RRID:Addgene_149639)
参考文献部分: A CRISPR interference platform for selective downregulation of gene expression in Borrelia burgdorferi. Takacs CN, Scott M, Chang Y, Kloos ZA, Irnov I, Rosa PA, Liu J, Jacobs-Wagner C. Appl Environ Microbiol. 2020 Nov 30. pii: AEM.02519-20. doi: 10.1128/AEM.02519-20. 10.1128/AEM.02519-20 PubMed 33257311

pBbdCas9Sarr2载体图谱质粒图谱和多克隆位点信息

pBbdCas9S_arr2质粒图谱

pBbdCas9Sarr2载体简介

pBbdCas9Sarr2载体序列

LOCUS       pBbdCas9S_arr2               11315 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  parental, all-in-one CRISPRi vector for B. burgdorferi, parental for
            gRNA cloning.
ACCESSION   .
VERSION     .
KEYWORDS    pBbdCas9S_arr2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11315)
  AUTHORS   Takacs CN, Scott M, Chang Y, Kloos ZA, Irnov I, Rosa PA, Liu J, 
            Jacobs-Wagner C
  TITLE     A CRISPR interference platform for selective downregulation of gene 
            expression in Borrelia burgdorferi.
  JOURNAL   Appl Environ Microbiol. 2020 Nov 30. pii: AEM.02519-20. doi: 
            10.1128/AEM.02519-20.
  PUBMED    33257311
REFERENCE   2  (bases 1 to 11315)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..11315
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1603..1647
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    1623..1639
                     /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)."
     protein_bind    1655..1671
                     /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)."
     RBS             1690..1701
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1708..5814
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TILERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5984..6775
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      6898..7486
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7387..7406
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaaaatttcc tggtaatcca acacaatctt atagaacttg tgaaccttta agaattgttg
       61 gtgttgttga agattgggaa ggacatcctg ttgaattaat aagaggaatg ttagattcat
      121 tagaagattt aaaacgtcgt ggtttacatg ttattgaaga ttaacctggc gcgccaaata
      181 atgcccgtgt ccctgttgaa tagggacacg gttttttaac aattttttta ttcctaggcc
      241 ttagtggccg gtacccgggg atccttatta ctggccgctt tctagcctgg atacttgtct
      301 ggctagttgc attagagagt cagctagagc cctagggcta gctgtttgtg tattcggagc
      361 cagtgtagtt tttcttttta ctaacgatac gggtaataat tgattgcctt ttacagcctg
      421 accttgagac aattgcaaaa gtctgtccac agaggtttga cctaataatc taaaatcttg
      481 tttgatagta gtaagtggag gtatgtagca actagaatct tctgtatcat cataacccac
      541 aactgaaata tcagccccaa ctctaagtcc actttcagta atagctcgca tagctcctaa
      601 agccatttga tcattagcaa ctaacatagc agtagggaca atgccttcat ttaacatttg
      661 cattgtttgc tgaaatcctg acatagcaga ccaatctcct tctctttctg ctattggttg
      721 tatttgattt cttgttaaat acttgtgcca tcctgctaat ctcaatctgg cacttactga
      781 acttaaggga cctgcaagta gtgcaatttg ttgatggcct aatgcaacaa ggtgttcaac
      841 tcctaacctt gttccatctt cgtgagaaaa aattatagag tttattgggg tttggtccga
      901 cacatccaaa aacaatgctg gaacattagt acatgctgct tcaactgcaa tcgcatcttg
      961 gtcatcaaga ggataattaa ttataagtcc actaacacgt tgagctaata aattatgtac
     1021 tgctgcctta caagcctcta cacctgatct ctcaaccatt gatacaacaa cgcttgcacc
     1081 aagttgatca gctctcgatt taattgcggc tacaatttga ctaggcgcat gtaatgcaag
     1141 tgatgatgtg gcaactccaa ttaaaagtga ttgtttcccc gcaagttgct gagcaacgcg
     1201 atttggtata taattcaact ctgccatagc tgcttcaacc ttctctcttg tttttgcaga
     1261 tacatgactt gcttggttca caactctcga aaccgtttgg tagctaaccc cggcatattc
     1321 agcaacatca tacaaagtaa caggtttcat ggatcattcc tccatgataa aatttaaatt
     1381 tctgactttg gcaaatcctt ttgcctgtta aaaagaatag cacaaaaatt aaaaaaaaag
     1441 aaacgcaaat agcaatagca ttaaaaataa ttattattat ttatttcttt gattaaaatc
     1501 aaaagccttt aatgtatttt cttgatcttt catgcgctta acggtaaaat ccaaggtatt
     1561 aagttcacct tccacttttc tctctctatc ttctctagaa aatcataaaa aatttatttg
     1621 ctttgtgagc ggataacaat tataatagat tcaattgtga gcggataaca atttcacaca
     1681 gaattcatta aagaggagaa attacatatg gataagaaat actcaatagg cttagctatc
     1741 ggcacaaata gcgtcggatg ggcggtgatc actgatgaat ataaggttcc gtctaaaaag
     1801 ttcaaggttc tgggaaatac agaccgccac agtatcaaaa aaaatcttat aggggctctt
     1861 ttatttgaca gtggagagac agcggaagcg actcgtctca aacggacagc tcgtagaagg
     1921 tatacacgtc ggaagaatcg tatttgttat ctacaggaga ttttttcaaa tgagatggcg
     1981 aaagtagatg atagtttctt tcatcgactt gaagagtctt ttttggtgga agaagacaag
     2041 aagcatgaac gtcatcctat ttttggaaat atagtagatg aagttgctta tcatgagaaa
     2101 tatccaacta tctatcatct gcgaaaaaaa ttggtagatt ctactgataa agcggatttg
     2161 cgcttaatct atttggcctt agcgcatatg attaagtttc gtggtcattt tttgattgag
     2221 ggagatttaa atcctgataa tagtgatgtg gacaaactat ttatccagtt ggtacaaacc
     2281 tacaatcaat tatttgaaga aaaccctatt aacgcaagtg gagtagatgc taaagcgatt
     2341 ctttctgcac gattgagtaa atcaagacga ttagaaaatc tcattgctca gctccccggt
     2401 gagaagaaaa atggcttatt tgggaatctc attgctttgt cattgggttt gacccctaat
     2461 tttaaatcaa attttgattt ggcagaagat gctaaattac agctttcaaa agatacttac
     2521 gatgatgatt tagataattt attggcgcaa attggagatc aatatgctga tttgtttttg
     2581 gcagctaaga atttatcaga tgctatttta ctttcagata tcctaagagt aaatactgaa
     2641 ataactaagg ctcccctatc agcttcaatg attaaacgct acgatgaaca tcatcaagac
     2701 ttgactcttt taaaagcttt agttcgacaa caacttccag aaaagtataa agaaatcttt
     2761 tttgatcaat caaaaaacgg atatgcaggt tatattgatg ggggagctag ccaagaagaa
     2821 ttttataaat ttatcaaacc aattttagaa aaaatggatg gtactgagga attattggtg
     2881 aaactaaatc gtgaagattt gctgcgcaag caacggacct ttgacaacgg ctctattccc
     2941 catcaaattc acttgggtga gctgcatgct attttgagaa gacaagaaga cttttatcca
     3001 tttttaaaag acaatcgtga gaagattgaa aaaatcttga cttttcgaat tccttattat
     3061 gttggtccat tggcgcgtgg caatagtcgt tttgcatgga tgactcggaa gtctgaagaa
     3121 acaattaccc catggaattt tgaagaagtt gtcgataaag gtgcttcagc tcaatcattt
     3181 attgaacgca tgacaaactt tgataaaaat cttccaaatg aaaaagtact accaaaacat
     3241 agtttgcttt atgagtattt tacggtttat aacgaattga caaaggtcaa atatgttact
     3301 gaaggaatgc gaaaaccagc atttctttca ggtgaacaga agaaagccat tgttgattta
     3361 ctcttcaaaa caaatcgaaa agtaaccgtt aagcaattaa aagaagatta tttcaaaaaa
     3421 atagaatgtt ttgatagtgt tgaaatttca ggagttgaag atagatttaa tgcttcatta
     3481 ggtacctacc atgatttgct aaaaattatt aaagataaag attttttgga taatgaagaa
     3541 aatgaagata tcttagagga tattgtttta acattgacct tatttgaaga tagggagatg
     3601 attgaggaaa gacttaaaac atatgctcac ctctttgatg ataaggtgat gaaacagctt
     3661 aaacgtcgcc gttatactgg ttggggacgt ttgtctcgaa aattgattaa tggtattagg
     3721 gataagcaat ctggcaaaac aatattagat tttttgaaat cagatggttt tgccaatcgc
     3781 aattttatgc agctgatcca tgatgatagt ttgacattta aagaagacat tcaaaaagca
     3841 caagtgtctg gacaaggcga tagtttacat gaacatattg caaatttagc tggtagccct
     3901 gctattaaaa aaggtatttt acagactgta aaagttgttg atgaattggt caaagtaatg
     3961 gggcggcata agccagaaaa tatcgttatt gaaatggcac gtgaaaatca gacaactcaa
     4021 aagggccaga aaaattcgcg agagcgtatg aaacgaatcg aagaaggtat caaagaatta
     4081 ggaagtcaga ttcttaaaga gcatcctgtt gaaaatactc aattgcaaaa tgaaaagctc
     4141 tatctctatt atctccaaaa tggaagagac atgtatgtgg accaagaatt agatattaat
     4201 cgtttaagtg attatgatgt cgatgccatt gttccacaaa gtttccttaa agacgattca
     4261 atagacaata aggtcttaac gcgttctgat aaaaatcgtg gtaaatcgga taacgttcca
     4321 agtgaagaag tagtcaaaaa gatgaaaaac tattggagac aacttctaaa cgccaagtta
     4381 atcactcaac gtaagtttga taatttaacg aaagctgaac gtggaggttt gagtgaactt
     4441 gataaagctg gttttatcaa acgccaattg gttgaaactc gccaaatcac taagcatgtg
     4501 gcacaaattt tggatagtcg catgaatact aaatacgatg aaaatgataa acttattcga
     4561 gaggttaaag tgattacctt aaaatctaaa ttagtttctg acttccgaaa agatttccaa
     4621 ttctataaag tacgtgagat taacaattac catcatgccc atgatgcgta tctaaatgcc
     4681 gtcgttggaa ctgctttgat taagaaatat ccaaaacttg aatcggagtt tgtctatggt
     4741 gattataaag tttatgatgt tcgtaaaatg attgctaagt ctgagcaaga aataggcaaa
     4801 gcaaccgcaa aatatttctt ttactctaat atcatgaact tcttcaaaac agaaattaca
     4861 cttgcaaatg gagagattcg caaacgccct ctaatcgaaa ctaatgggga aactggagaa
     4921 attgtctggg ataaagggcg agattttgcc acagtgcgca aagtattgtc catgccccaa
     4981 gtcaatattg tcaagaaaac agaagtccag acaggcggat tctccaagga gtcaatttta
     5041 ccaaaaagaa attcggacaa gcttattgct cgtaaaaaag actgggatcc aaaaaaatat
     5101 ggtggttttg atagtccaac ggtagcttat tcagtcctag tggttgctaa ggtggaaaaa
     5161 gggaaatcga agaagttaaa atccgttaaa gagttactag ggatcacaat tctggaaaga
     5221 agttcctttg aaaaaaatcc gattgacttt ttagaagcta aaggatataa ggaagttaaa
     5281 aaagacttaa tcattaaact acctaaatat agtctttttg agttagaaaa cggtcgtaaa
     5341 cggatgctgg ctagtgccgg agaattacaa aaaggaaatg agctggctct gccaagcaaa
     5401 tatgtgaatt ttttatattt agctagtcat tatgaaaagt tgaagggtag tccagaagat
     5461 aacgaacaaa aacaattgtt tgtggagcag cataagcatt atttagatga gattattgag
     5521 caaatcagtg aattttctaa gcgtgttatt ttagcagatg ccaatttaga taaagttctt
     5581 agtgcatata acaaacatag agacaaacca atacgtgaac aagcagaaaa tattattcat
     5641 ttatttacgt tgacgaatct tggagctccc gctgctttta aatattttga tacaacaatt
     5701 gatcgtaaac gatatacgtc tacaaaagaa gttttagatg ccactcttat ccatcaatcc
     5761 atcactggtc tttatgaaac acgcattgat ttgagtcagc taggaggtga ctaataaaag
     5821 cttgatcaga tctcagcttt tttttgaagt gcctggcagt aagttgaaaa aatttgaaat
     5881 aaacttggac tatgttaata aaaaaataca aattaacgaa caaaaagatt tgaattatta
     5941 tataatttag ttaaaagcaa ttttaaatga gggaggtttc catatgaggg aagcggtgat
     6001 cgccgaagta tcgactcaac tatcagaggt agttggcgtc atcgagcgcc atctcgaacc
     6061 gacgttgctg gccgtacatt tgtacggctc cgcagtggat ggcggcctga agccacacag
     6121 tgatattgat ttgctggtta cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt
     6181 gatcaacgac cttttggaaa cttcggcttc ccctggagag agcgagattc tccgcgctgt
     6241 agaagtcacc attgttgtgc acgacgacat cattccgtgg cgttatccag ctaagcgcga
     6301 actgcaattt ggagaatggc agcgcaatga cattcttgca ggtatcttcg agccagccac
     6361 gatcgacatt gatctggcta tcttgctgac aaaagcaaga gaacatagcg ttgccttggt
     6421 aggtccagcg gcggaggaac tctttgatcc ggttcctgaa caggatctat ttgaggcgct
     6481 aaatgaaacc ttaacgctat ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt
     6541 agtgcttacg ttgtcccgca tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga
     6601 tgtcgctgcc gactgggcaa tggagcgcct gccggcccag tatcagcccg tcatacttga
     6661 agctagacag gcttatcttg gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt
     6721 ggaagaattt gtccactacg tgaaaggcga gatcaccaag gtagtcggca aataatgacg
     6781 tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcatgacca aaatccctta
     6841 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6901 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6961 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     7021 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     7081 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     7141 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     7201 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     7261 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     7321 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     7381 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     7441 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     7501 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcact
     7561 agatccccgg caagttcatc cttagggttt tcatatctct taaaaagtca atttctgctt
     7621 gattaatttc ttgtggatta ttgtttttgc ggtttttcat tttttatctc cttgagtatt
     7681 gaattttttc ttatatataa atatatagca aaaactattt ttgccaactt ttttacaaaa
     7741 aattttttat taaaacactt agggctttgc taaattctct tttaaaagaa cttagtaaag
     7801 ccctatggat ttaagaactg attatactta cgtagtaaaa atactacaga tattaatagt
     7861 atagaccata tcaaaatcat aatcaattta tatttatcta aaattgtagt ccatacaaaa
     7921 tacaaaaaaa ttaggttttt ctaaattcta ttttaaaaga acttagtaac acctaatctc
     7981 taaattactt acaaaattct aaaattagat ttttagttct tcatatttct ttaaaagttt
     8041 tttaagaaaa tctttttgat tttcaaaaat ttcatgcatc ataaaactag taaatttgga
     8101 tttttttttg taaaaatcat aactttcttg ggttttaagt tgaaatctta atggttttat
     8161 tggattttgc ctagattttt ttaacgccgg actttcttta tcttttataa aattataaga
     8221 atttttaata ccattttcta ttaaataatc ttcttttata agcccctctt gcagtgcagt
     8281 tgctaatctt aaatacttat acgcttgagt ttttgctaca tcaaagtctt ttataaaatc
     8341 aagaaaagtt ttataaccat caaatttata atattgatta tcttttattt cttttaaaat
     8401 tttaatattt tccaccctgt ggaatatttc ttttttgaaa ttgtcattta acttttcctt
     8461 tagtgtcttg taataagaaa ttttactttc attaatcatt aaattttgtt caacataagt
     8521 atctatcttt tttgttaaaa ctttaacaac acgttttttt tccataaaaa ttttccttta
     8581 tttttgcaac attttcctta attgatataa ttgttactat tttagatttt ttaatatcca
     8641 cttaagaatt aatccctcac tagaatattt tttattatag aattcatata catgtttttc
     8701 taaatctata aatttatcaa caattgcttt attatagggt gtatctatct tttcttttct
     8761 taataatcgt ctcatattct ttaaataaca acaaacactg ccggttttaa acctaaattc
     8821 tatgtaatat atttttgaaa aagtatatac attttcaacc ccgtttattt catactttac
     8881 aaaaacatta tttattggtt ttctataacc atagtaaatg cctaaaaatt tgtccccgtc
     8941 tttcatagga tatagataaa gtttttgaat ctctgattta tctaataatt tttcaaaaaa
     9001 aattatacac cgcttttgtt tgttactaac tactctatgc aatgccttta atatttttgt
     9061 gtgatatata tttttatcac cttttttttc tttcagcagg aagatacttt cttttggaaa
     9121 tttaattttt tgttcattta ttcttgcagc cttaagtccg attttcatat ttaatcctta
     9181 tatagccttt ctatttaatt cttttaaact caataaactt ttttggtttt tcattagttc
     9241 taataattcg caataatatg tgcaaaacac tttattgtat tctattttct tttggttatt
     9301 caaatattct tttataattg gctttagaag ttcaatattt acttcttttt ttagttgttc
     9361 aataaggata ttgaaaatgt ttgtttttaa atcttgataa ttttgttgag aattttcttt
     9421 ttttctttca attgacttct ctaatttacg ctttatattg tttaaatcgc tatatttatg
     9481 actttcaata ataaaatgag gcttaaattt gtaattttcg tatatttttt ggaaacttgt
     9541 ttctaattgt ttagaattat attcattttt ttcaaattct ttttgagtgt tgcatagaat
     9601 ttcctttaat ttttcttgtt tttctttcat acaagattta gtaaaatgta tatttttatt
     9661 ttttagtaat ttaatttcta ttatttttat gattttaagt attttgatta tattatcttt
     9721 atcaatattt aaattgaata aaattgtaag aaaaattgaa agaatttttt tacataaaaa
     9781 gttacattta ttgaagtaat ttcttacttg atacttttct atttcattaa tctttctttc
     9841 ttcttttata ttatttttat tatttaaaca ctcctttaaa tctacactcc catttttagt
     9901 aaatctatct ttaaggcccc ttgaaactct attttgaaat cttaaatctt ttttttcttg
     9961 aaagtattta tagatttttt ggtaacattc tttttttgaa aaattaagtt gatagtatat
    10021 ttcggtcccc aaatttaccc ctaagtgttt gtgataatta gttgtgactt taaattcttt
    10081 ttctaatttg taaaggtaat tttgcagtgt ttttagttta acgggtgttt gactatttct
    10141 tttaaggttt tcattaaagt aatagagtat gtttttttgt gtgtatttct tatatgtttt
    10201 atttatgtac gctagtgttg agataaggac tactaattta tactgtaatt tattgtagca
    10261 agttgtggtt ttactggttt tgtttatatt agataagttt tccatttata gctccttgaa
    10321 taccggattt tttatctata aagtatatat atcacaattt tgatttaaaa gtaaattctt
    10381 tttttaaaaa tacaaaaatt ttttataaat tatatagacc atgatattgt attaatacta
    10441 attgatttta atttaaggtt gaactaaaat aaacatagtt ttgtagaaaa taatttttca
    10501 ttattttgct acttgatatt ggatcgtaaa aatattaggg ctttacaaag ttcttttaaa
    10561 agagaattta gtaaagccct aagtctttta ataaaaaatt ttttgtaaaa aagttggcaa
    10621 aaatagtttt tgctatatat ttatatataa gaaaaaattc aatactcaag gagataaaaa
    10681 atggaaaact taaacaataa tccacaaaaa attaatcaaa taaataagca aactaatttt
    10741 attggttttg aaaattttat agaacttgat aatcaagtta atcaaactaa agaaaaaagt
    10801 aaattaagta agataggtaa gaaattaccc ggaataagca gtcaagtcta gtgcggccgc
    10861 atggcttgtt atgactgttt ttttggggta cagtctatgc ctcgggcatc caagcagcaa
    10921 gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt tacgcagcag
    10981 ggcagtcgcc ctaaaacaaa gttaaacatc atgaggaggt aacgtcatgg taaaagattg
    11041 gattcctatt tctcatgata attataaaca agtacaagga ccattttatc atggaactaa
    11101 agctaattta gcaattggtg atttactaac tacaggattt atttctcatt ttgaagatgg
    11161 tcgaattctt aaacatattt atttttcagc tttaatggaa ccagcagttt ggggagctga
    11221 acttgctatg tcactatctg gtcttgaagg tcgtggttat atatatatag ttgaaccaac
    11281 aggaccattt gaagatgatc caaatcttac aaata
//


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