pCasKP-apr

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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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pCasKPapr载体基本信息

质粒名称: pCasKP-apr 
出品公司: 爱迪基因
目录编号: 117231
存储实验室: Quanjiang Ji

载体骨架基本信息
载体名称: --
载体出品公司: --
载体抗性: 安普霉素
空载体大小: --
完整质粒大小: 11409 bp
载体修饰: --
载体类型: 细菌表达,CRISPR,合成生物学
筛选标记: --
高拷贝/低拷贝: 低拷贝
生长条件: 30 ℃

插入基因信息
插入基因名称: Cas9
物种来源: --
插入大小: --
基因突变: --
GenBank编号 --
启动子 --
标签/融合蛋白: --
克隆方法: Gibson克隆
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: pCasKP-apr was a gift from Quanjiang Ji (Addgene plasmid # 117231 ; http://n2t.net/addgene:117231 ; RRID:Addgene_117231)
参考文献部分: Precise and efficient genome editing in Klebsiella pneumoniae using CRISPR-Cas9 and CRISPR-assisted cytidine deaminase. Wang Y, Wang S, Chen W, Song L, Zhang Y, Shen Z, Yu F, Li M, Ji Q. Appl Environ Microbiol. 2018 Sep 14. pii: AEM.01834-18. doi: 10.1128/AEM.01834-18. 10.1128/AEM.01834-18 PubMed 30217854

pCasKPapr载体图谱质粒图谱和多克隆位点信息

pCasKP-apr质粒图谱

pCasKPapr载体简介

pCasKPapr载体序列

LOCUS       pCasKP-apr             11393 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Bacterial expression of Cas9 nuclease and lambda-Red system in 
            Klebsiella Pneumoniae.
ACCESSION   .
VERSION     .
KEYWORDS    pCasKP-apr
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11393)
  AUTHORS   Wang Y, Wang S, Chen W, Song L, Zhang Y, Shen Z, Yu F, Li M, Ji Q
  TITLE     Precise and efficient genome editing in Klebsiella pneumoniae using 
            CRISPR-Cas9 and CRISPR-assisted cytidine deaminase.
  JOURNAL   Appl Environ Microbiol. 2018 Sep 14. pii: AEM.01834-18. doi: 
            10.1128/AEM.01834-18.
  PUBMED    30217854
REFERENCE   2  (bases 1 to 11393)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..11393
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             344..760
                     /codon_start=1
                     /gene="gam"
                     /product="inhibitor of the host RecBCD nuclease in the 
                     lambda Red system"
                     /label=Gam
                     /note="protects the double-stranded DNA substrate for 
                     recombination"
                     /translation="MDINTETEIKQKHSLTPFPVFLISPAFRGRYFHSYFRSSAMNAYY
                     IQDRLEAQSWARHYQQLAREEKEAELADDMEKGLPQHLFESLCIDHLQRHGASKKSITR
                     AFDDDVEFQERMAEHIRYMVETIAHHQVDIDSEV"
     CDS             766..1551
                     /codon_start=1
                     /gene="bet"
                     /product="single-stranded DNA binding recombinase in the 
                     lambda Red system"
                     /label=Beta
                     /note="mediates pairing of a single-stranded DNA with a 
                     complementary target"
                     /translation="MSTALATLAGKLAERVGMDSVDPQELITTLRQTAFKGDASDAQFI
                     ALLIVANQYGLNPWTKEIYAFPDKQNGIVPVVGVDGWSRIINENQQFDGMDFEQDNESC
                     TCRIYRKDRNHPICVTEWMDECRREPFKTREGREITGPWQSHPKRMLRHKAMIQCARLA
                     FGFAGIYDKDEAERIVENTAYTAERQPERDITPVNDETMQEINTLLIALDKTWDDDLLP
                     LCSQIFRRDIRASSELTQAEAVKALGFLKQKAAEQKVAA"
     CDS             1548..2228
                     /codon_start=1
                     /gene="exo"
                     /product="5' to 3' double-stranded DNA exonuclease in the 
                     lambda Red system"
                     /label=Exo
                     /note="creates 3' single-stranded DNA tails"
                     /translation="MTPDIILQRTGIDVRAVEQGEDAWHKLRLGVITASEVHNVIAKPR
                     SGKKWPDMKMSYFHTLLAEVCTGVAPEVNAKALAWGKQYENDARTLFEFTSGVNVTESP
                     IIYRDESMRTACSPDGLCSDGNGLELKCPFTSRDFMKFRLGGFEAIKSAYMAQVQYSMW
                     VTRKNAWYFANYDPRMKREGLHYVVIERDEKYMASFDEIVPEFIEKMDEALAEIGFVFG
                     EQWR"
     CDS             2810..6916
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=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             complement(7051..8001)
                     /codon_start=1
                     /gene="rep101 encoding an A56V mutation"
                     /product="temperature-sensitive version of the RepA protein
                     needed for replication with the pSC101 origin (Armstrong et
                     al., 1984)"
                     /label=Rep101(Ts)
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      8049..8271
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     promoter        8901..9003
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(9070..9089)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             9459..10259
                     /codon_start=1
                     /gene="aac(3)IV"
                     /product="aminoglycoside 3-N-acetyltransferase type IV     
                               "
                     /label=ApmR
                     /note="confers resistance to apramycin (Paget and Davies, 
                     1996)"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     CDS             complement(10489..11367)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg tttttttggg
      301 ctagcgaaaa gatgtttcgt gaagccgtcg acgcttataa aaaatggata ttaatactga
      361 aactgagatc aagcaaaagc attcactaac cccctttcct gttttcctaa tcagcccggc
      421 atttcgcggg cgatattttc acagctattt caggagttca gccatgaacg cttattacat
      481 tcaggatcgt cttgaggctc agagctgggc gcgtcactac cagcagctcg cccgtgaaga
      541 gaaagaggca gaactggcag acgacatgga aaaaggcctg ccccagcacc tgtttgaatc
      601 gctatgcatc gatcatttgc aacgccacgg ggccagcaaa aaatccatta cccgtgcgtt
      661 tgatgacgat gttgagtttc aggagcgcat ggcagaacac atccggtaca tggttgaaac
      721 cattgctcac caccaggttg atattgattc agaggtataa aacgaatgag tactgcactc
      781 gcaacgctgg ctgggaagct ggctgaacgt gtcggcatgg attctgtcga cccacaggaa
      841 ctgatcacca ctcttcgcca gacggcattt aaaggtgatg ccagcgatgc gcagttcatc
      901 gcattactga tcgttgccaa ccagtacggc cttaatccgt ggacgaaaga aatttacgcc
      961 tttcctgata agcagaatgg catcgttccg gtggtgggcg ttgatggctg gtcccgcatc
     1021 atcaatgaaa accagcagtt tgatggcatg gactttgagc aggacaatga atcctgtaca
     1081 tgccggattt accgcaagga ccgtaatcat ccgatctgcg ttaccgaatg gatggatgaa
     1141 tgccgccgcg aaccattcaa aactcgcgaa ggcagagaaa tcacggggcc gtggcagtcg
     1201 catcccaaac ggatgttacg tcataaagcc atgattcagt gtgcccgtct ggccttcgga
     1261 tttgctggta tctatgacaa ggatgaagcc gagcgcattg tcgaaaatac tgcatacact
     1321 gcagaacgtc agccggaacg cgacatcact ccggttaacg atgaaaccat gcaggagatt
     1381 aacactctgc tgatcgccct ggataaaaca tgggatgacg acttattgcc gctctgttcc
     1441 cagatatttc gccgcgacat tcgtgcatcg tcagaactga cacaggccga agcagtaaaa
     1501 gctcttggat tcctgaaaca gaaagccgca gagcagaagg tggcagcatg acaccggaca
     1561 ttatcctgca gcgtaccggg atcgatgtga gagctgtcga acagggggaa gatgcgtggc
     1621 acaaattacg gctcggcgtc atcaccgctt cagaagttca caacgtgata gcaaaacccc
     1681 gctccggaaa gaagtggcct gacatgaaaa tgtcctactt ccacaccctg cttgctgagg
     1741 tttgcaccgg tgtggctccg gaagttaacg ctaaagcact ggcctgggga aaacagtacg
     1801 agaacgacgc cagaaccctg tttgaattca cttccggcgt gaatgttact gaatccccga
     1861 tcatctatcg cgacgaaagt atgcgtaccg cctgctctcc cgatggttta tgcagtgacg
     1921 gcaacggcct tgaactgaaa tgcccgttta cctcccggga tttcatgaag ttccggctcg
     1981 gtggtttcga ggccataaag tcagcttaca tggcccaggt gcagtacagc atgtgggtga
     2041 cgcgaaaaaa tgcctggtac tttgccaact atgacccgcg tatgaagcgt gaaggcctgc
     2101 attatgtcgt gattgagcgg gatgaaaagt acatggcgag ttttgacgag atcgtgccgg
     2161 agttcatcga aaaaatggac gaggcactgg ctgaaattgg ttttgtattt ggggagcaat
     2221 ggcgatgacg catcctcacg ataataagct tcctgctgaa catcaaaggc aagaaaacat
     2281 ctgttgtcaa agacagcatc cttgaacaag gacaattaac agttaacaaa taaaaacgca
     2341 aaagaaaatg ccgatattga ctaccggaag cagtgtgacc gtgtgcttct caaatgcctg
     2401 attcaggctg tctatgtgtg actgttgagc tgtaacaagt tgtctcaggt gttcaatttc
     2461 atgttctagt tgctttgttt tactggtttc acctgttcta ttaggtgtta catgctgttc
     2521 atctgttaca ttgtcgatct gttcatggtg aacagcttta aatgcaccaa aaactcgtaa
     2581 aagctctgat gtatctatct tttttacacc gttttcatct gtgcatatag ctatactgat
     2641 ttcgtcagac tcacagtcaa acatgccggt cagttggcct ggtgatggcg ggatcgttgt
     2701 atatttcttg acaccttttc ggcaccgccc taaaattctg cgtcctcata atatatgagg
     2761 cgatttatta cgtgtttacg aagcaaaagc taaaaccagg agctatttaa tggataagaa
     2821 atactcaata ggcttagata tcggcacaaa tagcgtcgga tgggcggtga tcactgatga
     2881 atataaggtt ccgtctaaaa agttcaaggt tctgggaaat acagaccgcc acagtatcaa
     2941 aaaaaatctt ataggggctc ttttatttga cagtggagag acagcggaag cgactcgtct
     3001 caaacggaca gctcgtagaa ggtatacacg tcggaagaat cgtatttgtt atctacagga
     3061 gattttttca aatgagatgg cgaaagtaga tgatagtttc tttcatcgac ttgaagagtc
     3121 ttttttggtg gaagaagaca agaagcatga acgtcatcct atttttggaa atatagtaga
     3181 tgaagttgct tatcatgaga aatatccaac tatctatcat ctgcgaaaaa aattggtaga
     3241 ttctactgat aaagcggatt tgcgcttaat ctatttggcc ttagcgcata tgattaagtt
     3301 tcgtggtcat tttttgattg agggagattt aaatcctgat aatagtgatg tggacaaact
     3361 atttatccag ttggtacaaa cctacaatca attatttgaa gaaaacccta ttaacgcaag
     3421 tggagtagat gctaaagcga ttctttctgc acgattgagt aaatcaagac gattagaaaa
     3481 tctcattgct cagctccccg gtgagaagaa aaatggctta tttgggaatc tcattgcttt
     3541 gtcattgggt ttgaccccta attttaaatc aaattttgat ttggcagaag atgctaaatt
     3601 acagctttca aaagatactt acgatgatga tttagataat ttattggcgc aaattggaga
     3661 tcaatatgct gatttgtttt tggcagctaa gaatttatca gatgctattt tactttcaga
     3721 tatcctaaga gtaaatactg aaataactaa ggctccccta tcagcttcaa tgattaaacg
     3781 ctacgatgaa catcatcaag acttgactct tttaaaagct ttagttcgac aacaacttcc
     3841 agaaaagtat aaagaaatct tttttgatca atcaaaaaac ggatatgcag gttatattga
     3901 tgggggagct agccaagaag aattttataa atttatcaaa ccaattttag aaaaaatgga
     3961 tggtactgag gaattattgg tgaaactaaa tcgtgaagat ttgctgcgca agcaacggac
     4021 ctttgacaac ggctctattc cccatcaaat tcacttgggt gagctgcatg ctattttgag
     4081 aagacaagaa gacttttatc catttttaaa agacaatcgt gagaagattg aaaaaatctt
     4141 gacttttcga attccttatt atgttggtcc attggcgcgt ggcaatagtc gttttgcatg
     4201 gatgactcgg aagtctgaag aaacaattac cccatggaat tttgaagaag ttgtcgataa
     4261 aggtgcttca gctcaatcat ttattgaacg catgacaaac tttgataaaa atcttccaaa
     4321 tgaaaaagta ctaccaaaac atagtttgct ttatgagtat tttacggttt ataacgaatt
     4381 gacaaaggtc aaatatgtta ctgaaggaat gcgaaaacca gcatttcttt caggtgaaca
     4441 gaagaaagcc attgttgatt tactcttcaa aacaaatcga aaagtaaccg ttaagcaatt
     4501 aaaagaagat tatttcaaaa aaatagaatg ttttgatagt gttgaaattt caggagttga
     4561 agatagattt aatgcttcat taggtaccta ccatgatttg ctaaaaatta ttaaagataa
     4621 agattttttg gataatgaag aaaatgaaga tatcttagag gatattgttt taacattgac
     4681 cttatttgaa gatagggaga tgattgagga aagacttaaa acatatgctc acctctttga
     4741 tgataaggtg atgaaacagc ttaaacgtcg ccgttatact ggttggggac gtttgtctcg
     4801 aaaattgatt aatggtatta gggataagca atctggcaaa acaatattag attttttgaa
     4861 atcagatggt tttgccaatc gcaattttat gcagctgatc catgatgata gtttgacatt
     4921 taaagaagac attcaaaaag cacaagtgtc tggacaaggc gatagtttac atgaacatat
     4981 tgcaaattta gctggtagcc ctgctattaa aaaaggtatt ttacagactg taaaagttgt
     5041 tgatgaattg gtcaaagtaa tggggcggca taagccagaa aatatcgtta ttgaaatggc
     5101 acgtgaaaat cagacaactc aaaagggcca gaaaaattcg cgagagcgta tgaaacgaat
     5161 cgaagaaggt atcaaagaat taggaagtca gattcttaaa gagcatcctg ttgaaaatac
     5221 tcaattgcaa aatgaaaagc tctatctcta ttatctccaa aatggaagag acatgtatgt
     5281 ggaccaagaa ttagatatta atcgtttaag tgattatgat gtcgatcaca ttgttccaca
     5341 aagtttcctt aaagacgatt caatagacaa taaggtctta acgcgttctg ataaaaatcg
     5401 tggtaaatcg gataacgttc caagtgaaga agtagtcaaa aagatgaaaa actattggag
     5461 acaacttcta aacgccaagt taatcactca acgtaagttt gataatttaa cgaaagctga
     5521 acgtggaggt ttgagtgaac ttgataaagc tggttttatc aaacgccaat tggttgaaac
     5581 tcgccaaatc actaagcatg tggcacaaat tttggatagt cgcatgaata ctaaatacga
     5641 tgaaaatgat aaacttattc gagaggttaa agtgattacc ttaaaatcta aattagtttc
     5701 tgacttccga aaagatttcc aattctataa agtacgtgag attaacaatt accatcatgc
     5761 ccatgatgcg tatctaaatg ccgtcgttgg aactgctttg attaagaaat atccaaaact
     5821 tgaatcggag tttgtctatg gtgattataa agtttatgat gttcgtaaaa tgattgctaa
     5881 gtctgagcaa gaaataggca aagcaaccgc aaaatatttc ttttactcta atatcatgaa
     5941 cttcttcaaa acagaaatta cacttgcaaa tggagagatt cgcaaacgcc ctctaatcga
     6001 aactaatggg gaaactggag aaattgtctg ggataaaggg cgagattttg ccacagtgcg
     6061 caaagtattg tccatgcccc aagtcaatat tgtcaagaaa acagaagtac agacaggcgg
     6121 attctccaag gagtcaattt taccaaaaag aaattcggac aagcttattg ctcgtaaaaa
     6181 agactgggat ccaaaaaaat atggtggttt tgatagtcca acggtagctt attcagtcct
     6241 agtggttgct aaggtggaaa aagggaaatc gaagaagtta aaatccgtta aagagttact
     6301 agggatcaca attatggaaa gaagttcctt tgaaaaaaat ccgattgact ttttagaagc
     6361 taaaggatat aaggaagtta aaaaagactt aatcattaaa ctacctaaat atagtctttt
     6421 tgagttagaa aacggtcgta aacggatgct ggctagtgcc ggagaattac aaaaaggaaa
     6481 tgagctggct ctgccaagca aatatgtgaa ttttttatat ttagctagtc attatgaaaa
     6541 gttgaagggt agtccagaag ataacgaaca aaaacaattg tttgtggagc agcataagca
     6601 ttatttagat gagattattg agcaaatcag tgaattttct aagcgtgtta ttttagcaga
     6661 tgccaattta gataaagttc ttagtgcata taacaaacat agagacaaac caatacgtga
     6721 acaagcagaa aatattattc atttatttac gttgacgaat cttggagctc ccgctgcttt
     6781 taaatatttt gatacaacaa ttgatcgtaa acgatatacg tctacaaaag aagttttaga
     6841 tgccactctt atccatcaat ccatcactgg tctttatgaa acacgcattg atttgagtca
     6901 gctaggaggt gactgatggc tggttggcgt actgttgtgg taaataccca tatggacagt
     6961 tttccctttg atatgtaacg gtgaacagtt gttctacttt tgtttgttag tcttgatgct
     7021 tcactgatag atacaagagc cataagaacc tcagatcctt ccgtatttag ccagtatgtt
     7081 ctctagtgtg gttcgttgtt tttgcgtgag ccatgagaac gaaccattga gatcatactt
     7141 actttgcatg tcactcaaaa attttgcctc aaaactggtg agctgaattt ttgcagttaa
     7201 agcatcgtgt agtgtttttc ttagtccgtt atgtaggtag gaatctgatg taatggttgt
     7261 tggtattttg tcaccattca tttttatctg gttgttctca agttcggtta cgagatccat
     7321 ttgtctatct agttcaactt ggaaaatcaa cgtatcagtc gggcggcctc gcttatcaac
     7381 caccaatttc atattgctgt aagtgtttaa atctttactt attggtttca aaacccattg
     7441 gttaagcctt ttaaactcat ggtagttatt ttcaagcatt aacatgaact taaattcatc
     7501 aaggctaatc tctatatttg ccttgtgagt tttcttttgt gttagttctt ttaataacca
     7561 ctcataaatc ctcatagagt atttgttttc aaaagactta acatgttcca gattatattt
     7621 tatgaatttt tttaactgga aaagataagg caatatctct tcactaaaaa ctaattctaa
     7681 tttttcgctt gagaacttgg catagtttgt ccactggaaa atctcaaagc ctttaaccaa
     7741 aggattcctg atttccacag ttctcgtcat cagctctctg gttgctttag ctaatacacc
     7801 ataagcattt tccctactga tgttcatcat ctgaacgtat tggttataag tgaacgatac
     7861 cgtccgttct ttccttgtag ggttttcaat cgtggggttg agtagtgcca cacagcataa
     7921 aattagcttg gtttcatgct ccgttaagtc atagcgacta atcgctagtt catttgcttt
     7981 gaaaacaact aattcagaca tacatctcaa ttggtctagg tgattttaat cactatacca
     8041 attgagatgg gctagtcaat gataattact agtccttttc ctttgagttg tgggtatctg
     8101 taaattctgc tagacctttg ctggaaaact tgtaaattct gctagaccct ctgtaaattc
     8161 cgctagacct ttgtgtgttt tttttgttta tattcaagtg gttataattt atagaataaa
     8221 gaaagaataa aaaaagataa aaagaataga tcccagccct gtgtataact cactacttta
     8281 gtcagttccg cagtattaca aaaggatgtc gcaaacgctg tttgctcctc tacaaaacag
     8341 accttaaaac cctaaaggct taagtagcac cctcgcaagc tcgggcaaat cgctgaatat
     8401 tccttttgtc tccgaccatc aggcacctga gtcgctgtct ttttcgtgac attcagttcg
     8461 ctgcgctcac ggctctggca gtgaatgggg gtaaatggca ctacaggcgc cttttatgga
     8521 ttcatgcaag gaaactaccc ataatacaag aaaagcccgt cacgggcttc tcagggcgtt
     8581 ttatggcggg tctgctatgt ggtgctatct gactttttgc tgttcagcag ttcctgccct
     8641 ctgattttcc agtctgacca cttcggatta tcccgtgaca ggtcattcag actggctaat
     8701 gcacccagta aggcagcggt atcatcaaca ggcttacccg tcttactgtc ggggatcgac
     8761 gctctccctt atgcgactcc tgcacctttc gtcttcgaat aaatacctgt gacggaagat
     8821 cacttcgcag aataaataaa tcctggtgtc cctgttgata ccgggaagcc ctgggccaac
     8881 ttttggcgaa aatgagacgt tgatcggcac gtaagaggtt ccaactttca ccataatgaa
     8941 ataagatcac taccgggcgt attttttgag ttatcgagat tttcaggagc taaggaagct
     9001 aaaatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa
     9061 gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg
     9121 ggtcatccac cggatcaatt cccctgctcg cgcaggctgg gtgccaagct ctcgggtaac
     9181 atcaaggccc gatccttgga gcccttgccc tcccgcacga tgatcgtgcc gtgatcgaaa
     9241 tccagatcct tgacccgcag ttgcaaaccc tcactgatcc ggctcacggt aactgatgcc
     9301 gtatttgcag taccagcgta cggcccacag aatgatgtca cgctgaaaat gccggccttt
     9361 gaatgggttc atgtgcagct ccatcagcaa aaggggatga taagtttatc accaccgact
     9421 atttgcaaca gtgccgttga tcgtgctatg atcgactgat gtcatcagcg gtggagtgca
     9481 atgtcgtgca atacgaatgg cgaaaagccg agctcatcgg tcagcttctc aaccttgggg
     9541 ttacccccgg cggtgtgctg ctggtccaca gctccttccg tagcgtccgg cccctcgaag
     9601 atgggccact tggactgatc gaggccctgc gtgctgcgct gggtccggga gggacgctcg
     9661 tcatgccctc gtggtcaggt ctggacgacg agccgttcga tcctgccacg tcgcccgtta
     9721 caccggacct tggagttgtc tctgacacat tctggcgcct gccaaatgta aagcgcagcg
     9781 cccatccatt tgcctttgcg gcagcggggc cacaggcaga gcagatcatc tctgatccat
     9841 tgcccctgcc acctcactcg cctgcaagcc cggtcgcccg tgtccatgaa ctcgatgggc
     9901 aggtacttct cctcggcgtg ggacacgatg ccaacacgac gctgcatctt gccgagttga
     9961 tggcaaaggt tccctatggg gtgccgagac actgcaccat tcttcaggat ggcaagttgg
    10021 tacgcgtcga ttatctcgag aatgaccact gctgtgagcg ctttgccttg gcggacaggt
    10081 ggctcaagga gaagagcctt cagaaggaag gtccagtcgg tcatgccttt gctcggttga
    10141 tccgctcccg cgacattgtg gcgacagccc tgggtcaact gggccgagat ccgttgatct
    10201 tcctgcatcc gccagaggcg ggatgcgaag aatgcgatgc cgctcgccag tcgattggct
    10261 gagctcatga gcggagaacg agatgacgtt ggaggggcaa ggtcgcgctg attgctgggg
    10321 caacacgtga aaggcgagat caccaaggta gtcggcaaat aatgtctaac ggtacccggg
    10381 gatcctctag agtcgacctg cataatgtgc ctgtcaaatg gacgaagcag ggattctgca
    10441 aaccctatgc tactccgtca agccgtcaat tgtctgattc gttaccaatt atgacaactt
    10501 gacggctaca tcattcactt tttcttcaca accggcacgg aactcgctcg ggctggcccc
    10561 ggtgcatttt ttaaataccc gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc
    10621 gacggtggcg ataggcatcc gggtggtgct caaaagcagc ttcgcctggc tgatacgttg
    10681 gtcctcgcgc cagcttaaga cgctaatccc taactgctgg cggaaaagat gtgacagacg
    10741 cgacggcgac aagcaaacat gctgtgcgac gctggcgata tcaaaattgc tgtctgccag
    10801 gtgatcgctg atgtactgac aagcctcgcg tacccgatta tccatcggtg gatggagcga
    10861 ctcgttaatc gcttccatgc gccgcagtaa caattgctca agcagattta tcgccagcag
    10921 ctccgaatag cgcccttccc cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa
    10981 atgcggctgg tgcgcttcat ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca
    11041 gttaagccat tcatgccagt aggcgcgcgg acgaaagtaa acccactggt gataccattc
    11101 gcgagcctcc ggatgacgac cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc
    11161 acccggtcgg caaacaaatt ctcgtccctg atttttcacc accccctgac cgcgaatggt
    11221 gagattgaga atataacctt tcattcccag cggtcggtcg ataaaaaaat cgagataacc
    11281 gttggcctca atcggcgtta aacccgccac cagatgggca ttaaacgagt atcccggcag
    11341 caggggatca ttttgcgctt cagccatact tttcatactc ccgccattca gag
//

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