pHSB04X

价格:1200元

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: pHSB04X 
出品公司: 爱迪基因
目录编号: 117258
存储实验室: Sheng Yang

载体骨架基本信息
载体名称: pCas
载体出品公司: Sheng Yang, Addgene Plasmid #62225
载体抗性: 卡那霉素低(25 ug/ml)
空载体大小: 4476 bp
完整质粒大小: --
载体修饰: --
载体类型: --
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, promotor PslpA-guide-RNA, homologous arms of Lb_1019,Cas9 (4107 bps), promoter PslpA-XbaI-guide-RNA (324 bps), homologous arm (1992 bps)
物种来源: Cas9 (Streptococcus pyogenes M1 GAS)
插入大小: --
基因突变: --
GenBank编号 Cas9:AE004092 (SPy_1046)
启动子 --
标签/融合蛋白: --
克隆方法: 限制性内切酶
5' 克隆位点: XbaI(未破坏)
3' 克隆位点: ApaI (未破坏)
5' 测序引物 --
3' 测序引物 --

文献引用方法
材料和方法部分: pHSB04X was a gift from Sheng Yang (Addgene plasmid # 117258 ; http://n2t.net/addgene:117258 ; RRID:Addgene_117258)
参考文献部分: Development of a RecE/T-Assisted CRISPR-Cas9 Toolbox for Lactobacillus. Huang H, Song X, Yang S. Biotechnol J. 2019 Jul;14(7):e1800690. doi: 10.1002/biot.201800690. Epub 2019 May 20. 10.1002/biot.201800690 PubMed 30927506

载体图谱质粒图谱和多克隆位点信息

pHSB04X质粒图谱

载体简介

载体序列

LOCUS       pHSB04X             14339 bp ds-DNA     circular SYN 06-JUN-2019
DEFINITION  Genome editing for Lactobacillus brevis ATCC367.
ACCESSION   .
VERSION     .
KEYWORDS    pHSB04X
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14339)
  TITLE     RecE/T-Assisted Genome Editing in Lactobacillus
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 14339)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 6, 2019 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..14339
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(319..1134)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1042..1061
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1126..1145
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1431..5537
                     /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"
     misc_RNA        complement(12358..12433)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             complement(12786..13736)
                     /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      13784..14006
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 ggtctgctat gtggtgctat ctgacttttt gctgttcagc agttcctgcc ctctgatttt
       61 ccagtctgac cacttcggat tatcccgtga caggtcattc agactggcta atgcacccag
      121 taaggcagcg gtatcatcaa cggggtctga cgctcagtgg aacgaaaact cacgttaagg
      181 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
      241 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt acgtttccaa
      301 ccaattaacc aattctgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
      361 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
      421 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
      481 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
      541 atcaccatga gtgacgactg aatccggtga gaatggcaaa agcttatgca tttctttcca
      601 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
      661 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca
      721 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
      781 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
      841 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
      901 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
      961 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
     1021 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     1081 gttggaattt aatcgcggcc tcgagcaaga cgtttcccgt tgaatatggc tcataacacc
     1141 ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc
     1201 ttgtgcaatg taacatcaga gattttgaga cacaacgtgg ctttcctgca gcgaaaagcc
     1261 ctgacaaccc ttgttcctaa aaaggaataa gcgttcggtc agtaaataat agaaataaaa
     1321 aatcagacct aagactgatg acaaaaagag aaaattttga taaaatagtc ttagaattaa
     1381 attaaaaagg gaggccaaat ataatgaaaa atatgaatga caatgatgtt atggataaga
     1441 aatactcaat aggcttagat atcggcacaa atagcgtcgg atgggcggtg atcactgatg
     1501 aatataaggt tccgtctaaa aagttcaagg ttctgggaaa tacagaccgc cacagtatca
     1561 aaaaaaatct tataggggct cttttatttg acagtggaga gacagcggaa gcgactcgtc
     1621 tcaaacggac agctcgtaga aggtatacac gtcggaagaa tcgtatttgt tatctacagg
     1681 agattttttc aaatgagatg gcgaaagtag atgatagttt ctttcatcga cttgaagagt
     1741 cttttttggt ggaagaagac aagaagcatg aacgtcatcc tatttttgga aatatagtag
     1801 atgaagttgc ttatcatgag aaatatccaa ctatctatca tctgcgaaaa aaattggtag
     1861 attctactga taaagcggat ttgcgcttaa tctatttggc cttagcgcat atgattaagt
     1921 ttcgtggtca ttttttgatt gagggagatt taaatcctga taatagtgat gtggacaaac
     1981 tatttatcca gttggtacaa acctacaatc aattatttga agaaaaccct attaacgcaa
     2041 gtggagtaga tgctaaagcg attctttctg cacgattgag taaatcaaga cgattagaaa
     2101 atctcattgc tcagctcccc ggtgagaaga aaaatggctt atttgggaat ctcattgctt
     2161 tgtcattggg tttgacccct aattttaaat caaattttga tttggcagaa gatgctaaat
     2221 tacagctttc aaaagatact tacgatgatg atttagataa tttattggcg caaattggag
     2281 atcaatatgc tgatttgttt ttggcagcta agaatttatc agatgctatt ttactttcag
     2341 atatcctaag agtaaatact gaaataacta aggctcccct atcagcttca atgattaaac
     2401 gctacgatga acatcatcaa gacttgactc ttttaaaagc tttagttcga caacaacttc
     2461 cagaaaagta taaagaaatc ttttttgatc aatcaaaaaa cggatatgca ggttatattg
     2521 atgggggagc tagccaagaa gaattttata aatttatcaa accaatttta gaaaaaatgg
     2581 atggtactga ggaattattg gtgaaactaa atcgtgaaga tttgctgcgc aagcaacgga
     2641 cctttgacaa cggctctatt ccccatcaaa ttcacttggg tgagctgcat gctattttga
     2701 gaagacaaga agacttttat ccatttttaa aagacaatcg tgagaagatt gaaaaaatct
     2761 tgacttttcg aattccttat tatgttggtc cattggcgcg tggcaatagt cgttttgcat
     2821 ggatgactcg gaagtctgaa gaaacaatta ccccatggaa ttttgaagaa gttgtcgata
     2881 aaggtgcttc agctcaatca tttattgaac gcatgacaaa ctttgataaa aatcttccaa
     2941 atgaaaaagt actaccaaaa catagtttgc tttatgagta ttttacggtt tataacgaat
     3001 tgacaaaggt caaatatgtt actgaaggaa tgcgaaaacc agcatttctt tcaggtgaac
     3061 agaagaaagc cattgttgat ttactcttca aaacaaatcg aaaagtaacc gttaagcaat
     3121 taaaagaaga ttatttcaaa aaaatagaat gttttgatag tgttgaaatt tcaggagttg
     3181 aagatagatt taatgcttca ttaggtacct accatgattt gctaaaaatt attaaagata
     3241 aagatttttt ggataatgaa gaaaatgaag atatcttaga ggatattgtt ttaacattga
     3301 ccttatttga agatagggag atgattgagg aaagacttaa aacatatgct cacctctttg
     3361 atgataaggt gatgaaacag cttaaacgtc gccgttatac tggttgggga cgtttgtctc
     3421 gaaaattgat taatggtatt agggataagc aatctggcaa aacaatatta gattttttga
     3481 aatcagatgg ttttgccaat cgcaatttta tgcagctgat ccatgatgat agtttgacat
     3541 ttaaagaaga cattcaaaaa gcacaagtgt ctggacaagg cgatagttta catgaacata
     3601 ttgcaaattt agctggtagc cctgctatta aaaaaggtat tttacagact gtaaaagttg
     3661 ttgatgaatt ggtcaaagta atggggcggc ataagccaga aaatatcgtt attgaaatgg
     3721 cacgtgaaaa tcagacaact caaaagggcc agaaaaattc gcgagagcgt atgaaacgaa
     3781 tcgaagaagg tatcaaagaa ttaggaagtc agattcttaa agagcatcct gttgaaaata
     3841 ctcaattgca aaatgaaaag ctctatctct attatctcca aaatggaaga gacatgtatg
     3901 tggaccaaga attagatatt aatcgtttaa gtgattatga tgtcgatcac attgttccac
     3961 aaagtttcct taaagacgat tcaatagaca ataaggtctt aacgcgttct gataaaaatc
     4021 gtggtaaatc ggataacgtt ccaagtgaag aagtagtcaa aaagatgaaa aactattgga
     4081 gacaacttct aaacgccaag ttaatcactc aacgtaagtt tgataattta acgaaagctg
     4141 aacgtggagg tttgagtgaa cttgataaag ctggttttat caaacgccaa ttggttgaaa
     4201 ctcgccaaat cactaagcat gtggcacaaa ttttggatag tcgcatgaat actaaatacg
     4261 atgaaaatga taaacttatt cgagaggtta aagtgattac cttaaaatct aaattagttt
     4321 ctgacttccg aaaagatttc caattctata aagtacgtga gattaacaat taccatcatg
     4381 cccatgatgc gtatctaaat gccgtcgttg gaactgcttt gattaagaaa tatccaaaac
     4441 ttgaatcgga gtttgtctat ggtgattata aagtttatga tgttcgtaaa atgattgcta
     4501 agtctgagca agaaataggc aaagcaaccg caaaatattt cttttactct aatatcatga
     4561 acttcttcaa aacagaaatt acacttgcaa atggagagat tcgcaaacgc cctctaatcg
     4621 aaactaatgg ggaaactgga gaaattgtct gggataaagg gcgagatttt gccacagtgc
     4681 gcaaagtatt gtccatgccc caagtcaata ttgtcaagaa aacagaagta cagacaggcg
     4741 gattctccaa ggagtcaatt ttaccaaaaa gaaattcgga caagcttatt gctcgtaaaa
     4801 aagactggga tccaaaaaaa tatggtggtt ttgatagtcc aacggtagct tattcagtcc
     4861 tagtggttgc taaggtggaa aaagggaaat cgaagaagtt aaaatccgtt aaagagttac
     4921 tagggatcac aattatggaa agaagttcct ttgaaaaaaa tccgattgac tttttagaag
     4981 ctaaaggata taaggaagtt aaaaaagact taatcattaa actacctaaa tatagtcttt
     5041 ttgagttaga aaacggtcgt aaacggatgc tggctagtgc cggagaatta caaaaaggaa
     5101 atgagctggc tctgccaagc aaatatgtga attttttata tttagctagt cattatgaaa
     5161 agttgaaggg tagtccagaa gataacgaac aaaaacaatt gtttgtggag cagcataagc
     5221 attatttaga tgagattatt gagcaaatca gtgaattttc taagcgtgtt attttagcag
     5281 atgccaattt agataaagtt cttagtgcat ataacaaaca tagagacaaa ccaatacgtg
     5341 aacaagcaga aaatattatt catttattta cgttgacgaa tcttggagct cccgctgctt
     5401 ttaaatattt tgatacaaca attgatcgta aacgatatac gtctacaaaa gaagttttag
     5461 atgccactct tatccatcaa tccatcactg gtctttatga aacacgcatt gatttgagtc
     5521 agctaggagg tgactgaagt atattttaga tgaagattat ttcttaatct aggctcgaga
     5581 tctatcgata agcttaagtg attagtcaaa gaatggtgat gacaattgta aattctattt
     5641 aatcactttg actagcaaat actaacaaca agacacacac accaaaaatc aaaaattcac
     5701 tacttttagt taaaaaccac gtaaccacaa gaactaatcc aatccatgta atcgggttct
     5761 tcaaatattt ctccaagatt ttcctcctct aatatgctca acttaaatga cctattcaat
     5821 aaatctatta tgctgctaaa tagtttatag gacaaataag tatactctaa tgacctataa
     5881 aagatagaaa attaaaaaat caagtgttcg cttcgctctc actgcccctc gacgttttag
     5941 tagcctttcc ctcacttcgt tcagtccaag ccaactaaaa gttttcgggc tactctctcc
     6001 ttctccccct aataattaat taaaatctta ctctgtatat ttctgctaat cattcgctaa
     6061 acagcaaaga aaaaacaaac acgtatcata gatataaatg taatggcata gtgcgggttt
     6121 tattttcagc ctgtatcata gctaaacaaa tcgagttgtg tgtccgtttt agggcgttct
     6181 gctagcttgt ttaaagtctc ttgaatgaat gtatgctcta agtcaaaaga atttgtcagc
     6241 gcctttatat agctttcttt ttcttctttt tttactttaa tgatcgatag caacaatgat
     6301 ttaacactag caagttgaat gccaccattt cttcctggtt taatcttaaa gaaaatttcc
     6361 tgattcgcct tcagtacctt cagcaattta tctaatgtcc gttcaggaat gcctagcact
     6421 tctctaatct cttttttggt cgtcactaaa taaggcttgt atacatcgct tttttcgcta
     6481 atataagcca ttaaatcttc tttccattct gacaaatgaa cacgttgacg ttcgcttctt
     6541 tttttcttga atttaaacca cccttgacgg acaaataaat ctttactggt taaatcactt
     6601 gatacccaag ctttgcaaag aatggtaatg tattccctat tagccccttg atagttttct
     6661 gaataggcac ttctaacaat tttgattact tctttttctt ctaagggttg atctaatcga
     6721 ttattaaact caaacatatt atattcgcac gtttcgattg aatagcctga actaaagtag
     6781 gctaaagaga gggtaaacat gacgttatta cgccctatta aacccttttc tcctgaaaat
     6841 ttcgtttcgt gcaataagag attaaaccag ggttcatcta cttgtttttt gccttctgta
     6901 ccgcttaaaa ccgttagact tgaacgagta aagcccttat tatctgtttg tttgaaagac
     6961 caatcttgcc attctttgaa agaataacgg taattaggat caaaaaattc tacattgtcc
     7021 gttcttggta tgcgagcaat accaaaatga ttacacgtta gatcaactgg caaagacttt
     7081 ccaaaatatt ctcggatatt ttgcgaaatt attttggctg ctttgacaga tttaaattct
     7141 gattttgaag tcacatagac tggcgtttct aaaacaaaat atgcttgata acctttatca
     7201 gatttgataa tcatagtagg cataaaacct aaatcaatag cggttgttaa aatatcgctt
     7261 gctgaaatag tttcttttgc cgtgtgaata tcaaaatcaa taaagaaggt attgatttgt
     7321 cttaaattgt tttcagaatg tcctttcgtg tatgaacggt tttcgtctgc atacgttcca
     7381 taacgataaa cgttgggtgt ccaatgtgta aatgtatctt gattttcttg aatcgcttcc
     7441 tcggaagtca gaacaacacc acgaccgcca atcatgcttg attttgagcg atacgcaaaa
     7501 atagcccctt tgcttttacc tggcttggta gtgattgagc gaattttact atttttaaat
     7561 ttgtacttta acaagccgtc atgaagcaca gtttctacaa caaaagggat attcattcag
     7621 ctgttctcct ttcctataaa tcctataaaa taggttgttt aattaacttg gtttgctttt
     7681 tcattcaact gtttcaatat tgcatgtttt gaaaaagatt tttttccttt ataagtcaat
     7741 ttttttccac taatcgaata aattattttg ttattttcta ttaacttata tatataatct
     7801 tccccctccg aagaaaaata cttatctgat tttgtttcta agtagatatt tctcttttct
     7861 aactctttct taaacgtttc tagtgtatag atatttgcta attttcttat ctccaataaa
     7921 ctatttttta tataagtttt acattcatca tgattcatac aaactccacc ttctataaat
     7981 gaatacaaaa aaagcaatca aacgatttcc gattgattgc ttaacaattc ttaaattcag
     8041 tagcttagat acttgaaaac tctctgattt ccctatataa tgatagtacg gttatatacc
     8101 gtcttcaaac aaagttaatt aaataacttc ttacgaggga agagttcatc tgactaactg
     8161 ataagcgttg gtttggcaat cttatcgggc tatgcattta taaaatgtcg tcaaacattt
     8221 tataaatgtg tcatggctct tttttcgttt ctattcagtt cgttgtttcg ttatatctag
     8281 tataccgctt ttaaaaaaaa taagcaacga tttcgtgcat tattcacacg aagtcattgc
     8341 ttttttcttc ttccatttct aaatccaatg ttacttgttc tgattctgtt tctggttctg
     8401 gttctgttgg ctcatttggg attaaatcca ctactagcgt tgagttagtt aactttgcaa
     8461 tttgttctag tgtttttatg gttggatctg attttcctga ttctattcgt gaataatttg
     8521 atctactcat ttctaattct tggggtaccg ccagcatttc ggaaaaaaac cacgctaagg
     8581 attttttcta taaaaagagc cgttatatta agaataaaac ggctctttta tacgtaaagg
     8641 acgtaaattc atttgcccag tgtcatgtaa tccttcaaat ttgtattctc caagaaaatt
     8701 gatatgttcc catcctaacg gccacgcata tggcattaaa tcttctctaa attctcctct
     8761 tgcttttaat tcttctacgg ctttttccat atatacagtg ttccacacac ttatagcgtt
     8821 aataattatg tttagtgcac tagctctttg taactggtct tggagagcac gttctctaaa
     8881 ttctccacgt tgtccaaaaa atataattct agctaatgca ttgattgctt ctcctttatt
     8941 taaacctttt tgaacccgtc tccttacggc tttattagat atgtaatcca gcgtaaagag
     9001 ggttttctcg attcgtccca tttctccaag tgctgttgcg agtttatttt gtcttgcata
     9061 tgatccgagc ttccccatga taagagcgct agggacctct ttagctcctt ggaagctgtc
     9121 agtagtatac ctaataattt atctacattc cctttagtaa cgtgtaactt tccaaattta
     9181 caaaagcgac tcatagaatt atttcctccc gttaaataat agataactat taaaaataga
     9241 caatacttgc tcataagtaa cggtacttaa attgtttact ttggcgtgtt tcattgcttg
     9301 atgaaactga tttttagtaa acagttgacg atattctcga ttgacccatt ttgaaacaaa
     9361 gtacgtatat agcttccaat atttatctgg aacatctgtg gtatggcggg taagttttat
     9421 taagacactg tttacttttg gtttaggatg aaagcattcc gctggcagct taagcaattg
     9481 ctgaatcgag acttgagtgt gcaagagcaa ccctagtgtt cggtgaatat ccaaggtacg
     9541 cttgtagaat ccttcttcaa caatcagata gatgtcagac gcatggcttt caaaaaccac
     9601 ttttttaata atttgtgtgc ttaaatggta aggaatactc ccaacaattt tatacctctg
     9661 tttgttaggg aattgaaact gtagaatatc ttggtgaatt aaagtgacac gagtattcag
     9721 ttttaatttt tctgacgata agttgaatag atgactgtct aattcaatag acgttacctg
     9781 tttacttatt ttagccagtt tcgtcgttaa atgcccttta cctgttccaa tttcgtaaac
     9841 ggtatcggtt tcttttaaat tcaattgttt tattatttgg ttgagtactt tttcactcgt
     9901 taaaaagttt tgagaatatt ttatattttt gttcatgtaa tcactccttc ttaattacaa
     9961 atttttagca tctaatttaa cttcaattcc tattatacaa aattttaaga tactgcacta
    10021 tcaacacact cttaagtttg cttctaagtc ttatttccat aacttctttt acgtttccgc
    10081 cattctttgc tgtttcgatt tttatgatat ggtgcaagtc agcacgaaca cgaaccgtct
    10141 tatctcccat tatatctttt tttgcactga ttggtgtatc atttcgtttt tctttttgtg
    10201 cgcttcttga taaaagggat agtaattcat tcctggttgc aaattttgaa aaccgctacg
    10261 gatcgcatct ttttctaaac tagggcccaa ccgcatcacg attaaacaga cgttcggact
    10321 aacttagtcg cgaccgtttt ttattggcgt ggtaaatcct atcatgacat gactgacagt
    10381 tatcgaaagt gaaagaagtc tagcgctgtt tgggcagtca gttattaaaa aaacgtagca
    10441 tttcaagaat ttcaggcgtt ttagtggctt ctaatgaatg ggaccagtaa actaacccta
    10501 atcattcaaa ataggagcaa aaataatgcg caaatttttg ggttttctgt tcttgctggc
    10561 gctaggggcc tgctttttct taccccaagt acaaactgag cttccaaaga ttctaaaaca
    10621 atcgcaattg accgttgccc aagcggctaa aaacatcggg gttcagttgc cgctggtcgg
    10681 ccgacaagag gcggctggtg ccacgccagt cgagtcgatt gttcgtccgc aaacgttaaa
    10741 gaaaacgtac tattatcatt tttcggcgga tgttccagta aaagttcaac aagtatttga
    10801 acgggccatc gctgtttata atgagaccgg tattgttgac ttagtggcag gtcaggccac
    10861 tgccgaacaa aatgaaatta cgatttttgt ttatcagaaa acgatgacgg ctaaacaagt
    10921 taattatttg gaattaggta agggtggtcc ggagattcag caactaacgg gatttcaagg
    10981 ttacacggct aatcatgcgc gtgcgggact gaacgtgaag tatggtcaaa gcattaagga
    11041 atctgtggcg atccacgaat tgggccatgc gttaggactc gatcacagtt cgagtcggca
    11101 atcagtgatg tatccaattg atcaggggct aacccagctg tctgctggtg atttaaaggc
    11161 actgaagtat ttgtattagg atgccgcgta aaaaaggcca cgacatgtca ttatcttttg
    11221 aggaccaata actaaaaaaa gcaattctga aaatgacttt ttcagaattg cttttttaat
    11281 tcaaatatta atttaagtcg aaattacttc gtcatgaaaa aaactccttt tcttttcgtc
    11341 tacaagaatc agtatactgc aaaggaagta atattcaagt gaaattttgc acaaaaagta
    11401 ttacgttttt cctaagatga acgaaaaaag cttatataac agcagtcatt ggttgatctg
    11461 aaaagtatat aaatgtgaag caaatgacta atcacttttt cagaagcaat attatcgtcc
    11521 gaatgcaacc ggttgttaat ttttcaaaag acagcgacta acagtgatga ataggtaatg
    11581 tttcagttaa atgtgagtgg gtagggagtt ttggcataca attatgagaa aatatgataa
    11641 aattaaatgt tgaaatgaga atcggacgtc cattgttagt gtttttgctg acagtgggtg
    11701 ccgcgaatta gccagaggag ttaacgtgta atgagccgaa ttttaattat tgaagatgaa
    11761 aaaaacctag cccggttcgt tgaactggag ttaaagcatg aagggtacga aaccaacgtg
    11821 cagtttaatg gtcgaaccgg cctggaagca gccttgtcac aggattttga tgtgattttg
    11881 ctagacctaa tgttgcctga attgaatggg atcgaagttg ctcgccgcgt gcgtgaggtt
    11941 aagagtacgc caatcattat gatgacggcg cgtgattctg tgattgatcg ggtttctgga
    12001 ttagatcatg gcgcagacga ctatatcgtt aagccatttg caattgaaga actgttagcg
    12061 cgtgttcggg cgctattgcg gcgaattcac ctagagggtg agcagaagag taccaaacag
    12121 acgacggtta agttcaagga tcttacaatt gaaaaggaaa accgcatcgt acgtcggggc
    12181 gacgaagtca ttaacctgac taaacgcgaa tatgaattgt tattagcctt gatggagaat
    12241 atcaatgttg ttttggcgcg agatgtcctg ttaaccaaag tgtggggtta tgaatcagaa
    12301 gtagaaacca acgtggttga tgtctacgtg cggtatcttc gcaagggccc aaaaaaagca
    12361 ccgactcggt gccacttttt caagttgata acggactagc cttattttaa cttgctattt
    12421 ctagctctaa aactgaaagt cggcgaaact aagtctagaa tagtataacc gaagttagcg
    12481 cttttgttaa gaaatttatt cgataacatc agcggacccg atcagctatc ccctaactac
    12541 gaagataagt atagcgattt ccttaaggag atacaaggat tacgccctga ttgtcgttgg
    12601 attgaaacaa ttatgggtat cctgtaatgt tattgttaca taactaaaag tctactaacc
    12661 tgattaaagc ctttgtaagt cgacgtatgg acagttttcc ctttgatatg taacggtgaa
    12721 cagttgttct acttttgttt gttagtcttg atgcttcact gatagataca agagccataa
    12781 gaacctcaga tccttccgta tttagccagt atgttctcta gtgtggttcg ttgtttttgc
    12841 gtgagccatg agaacgaacc attgagatca tacttacttt gcatgtcact caaaaatttt
    12901 gcctcaaaac tggtgagctg aatttttgca gttaaagcat cgtgtagtgt ttttcttagt
    12961 ccgttatgta ggtaggaatc tgatgtaatg gttgttggta ttttgtcacc attcattttt
    13021 atctggttgt tctcaagttc ggttacgaga tccatttgtc tatctagttc aacttggaaa
    13081 atcaacgtat cagtcgggcg gcctcgctta tcaaccacca atttcatatt gctgtaagtg
    13141 tttaaatctt tacttattgg tttcaaaacc cattggttaa gccttttaaa ctcatggtag
    13201 ttattttcaa gcattaacat gaacttaaat tcatcaaggc taatctctat atttgccttg
    13261 tgagttttct tttgtgttag ttcttttaat aaccactcat aaatcctcat agagtatttg
    13321 ttttcaaaag acttaacatg ttccagatta tattttatga atttttttaa ctggaaaaga
    13381 taaggcaata tctcttcact aaaaactaat tctaattttt cgcttgagaa cttggcatag
    13441 tttgtccact ggaaaatctc aaagccttta accaaaggat tcctgatttc cacagttctc
    13501 gtcatcagct ctctggttgc tttagctaat acaccataag cattttccct actgatgttc
    13561 atcatctgaa cgtattggtt ataagtgaac gataccgtcc gttctttcct tgtagggttt
    13621 tcaatcgtgg ggttgagtag tgccacacag cataaaatta gcttggtttc atgctccgtt
    13681 aagtcatagc gactaatcgc tagttcattt gctttgaaaa caactaattc agacatacat
    13741 ctcaattggt ctaggtgatt ttaatcacta taccaattga gatgggctag tcaatgataa
    13801 ttactagtcc ttttcctttg agttgtgggt atctgtaaat tctgctagac ctttgctgga
    13861 aaacttgtaa attctgctag accctctgta aattccgcta gacctttgtg tgtttttttt
    13921 gtttatattc aagtggttat aatttataga ataaagaaag aataaaaaaa gataaaaaga
    13981 atagatccca gccctgtgta taactcacta ctttagtcag ttccgcagta ttacaaaagg
    14041 atgtcgcaaa cgctgtttgc tcctctacaa aacagacctt aaaaccctaa aggcttaagt
    14101 agcaccctcg caagctcggt tgcggccgca atcgggcaaa tcgctgaata ttccttttgt
    14161 ctccgaccat caggcacctg agtcgctgtc tttttcgtga cattcagttc gctgcgctca
    14221 cggctctggc agtgaatggg ggtaaatggc actacaggcg ccttttatgg attcatgcaa
    14281 ggaaactacc cataatacaa gaaaagcccg tcacgggctt ctcagggcgt tttatggcg
//

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