pJK-caCas9-NatMX-Neut5L-Ade2 drive

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: pJK-caCas9-NatMX-Neut5L-Ade2 drive 
出品公司: 爱迪基因
目录编号: 89577
存储实验室: George Church

载体骨架基本信息
载体名称: pJK1027
载体出品公司: --
载体抗性: 氨苄青霉素
空载体大小: 7000 bp
完整质粒大小: --
载体修饰: 插入caCas9和Neut5L同源臂
载体类型: 酵母表达,合成生物学
筛选标记: Nourseothricin
高拷贝/低拷贝: 高拷贝
生长条件: 30 ℃

插入基因信息
插入基因名称: Ade2 gene drive
物种来源: C. albicans
插入大小: 1000 bp
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 TTGCAATTATTAGAGATCCAGAAAA
3' 测序引物 --

文献引用方法
材料和方法部分: pJK-caCas9-NatMX-Neut5L-Ade2 drive was a gift from George Church (Addgene plasmid # 89577 ; http://n2t.net/addgene:89577 ; RRID:Addgene_89577)
参考文献部分: A CRISPR-Cas9-based gene drive platform for genetic interaction analysis in Candida albicans. Shapiro RS, Chavez A, Porter CBM, Hamblin M, Kaas CS, DiCarlo JE, Zeng G, Xu X, Revtovich AV, Kirienko NV, Wang Y, Church GM, Collins JJ. Nat Microbiol. 2018 Jan;3(1):73-82. doi: 10.1038/s41564-017-0043-0. Epub 2017 Oct 23. 10.1038/s41564-017-0043-0 PubMed 29062088

载体图谱质粒图谱和多克隆位点信息

pJK-caCas9-NatMX-Neut5L-Ade2 drive质粒图谱

载体简介

载体序列

LOCUS        pJK-caCas9-NatMX-Neut5L-Ade2 drive               12969 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  caCas9 integrating vector into the Neut5L locus with gene drive for 
            targeting Ade2 locus.
ACCESSION   .
VERSION     .
KEYWORDS    pJK-caCas9-NatMX-Neut5L-Ade2 drive
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12969)
  AUTHORS   Shapiro RS, Chavez A, Porter CBM, Hamblin M, Kaas CS, DiCarlo JE, 
            Zeng G, Xu X, Revtovich AV, Kirienko NV, Wang Y, Church GM, Collins 
            JJ
  TITLE     A CRISPR-Cas9-based gene drive platform for genetic interaction 
            analysis in Candida albicans.
  JOURNAL   Nat Microbiol. 2018 Jan;3(1):73-82. doi: 10.1038/s41564-017-0043-0. 
            Epub 2017 Oct 23.
  PUBMED    29062088
REFERENCE   2  (bases 1 to 12969)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..12969
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             1..4104
                     /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             4117..4137
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     4142..4158
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     4143..4159
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      5095..5292
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        complement(5893..6236)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     primer_bind     complement(7199..7218)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7200..7218)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7225..7242)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7225..7241)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7234..7256)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(7470..7489)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      7742..7761
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     misc_RNA        complement(7766..7840)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        complement(7866..7940)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     9587..9608
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9771..9875
                     /gene="bla"
                     /label=AmpR promoter
     CDS             9876..10736
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(10094..10113)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10907..11495
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11396..11415
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11649..11666
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11783..11804
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        11819..11849
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11857..11873
                     /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     11862..11884
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11881..11897
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11881..11897
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        11918..11936
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 atggataaaa aatactccat aggtttggac attggaacca attcagttgg atgggctgta
       61 atcactgatg aatataaggt tccttctaaa aagttcaaag ttttaggaaa tactgataga
      121 cactccatta aaaagaattt aattggtgcc ttgttgtttg acagtggtga aacagctgaa
      181 gcaactcgtt tgaaaagaac agctagaaga agatatacca gaagaaagaa cagaatttgt
      241 tacttgcaag aaatcttttc taatgaaatg gcaaaggttg atgattcttt ttttcataga
      301 ttggaagaaa gtttcttagt tgaagaagat aagaaacatg aaagacatcc aatctttggt
      361 aatattgtag atgaagtggc ttaccacgaa aaatatccta caatttatca tttgagaaaa
      421 aaattagttg actccactga taaagctgac ttaagactta tatatttggc attggctcac
      481 atgatcaagt tcagaggtca cttcttaatt gaaggtgatt tgaatccaga taattccgat
      541 gtcgacaagt tgtttattca attggttcaa acttacaacc aacttttcga ggaaaatcca
      601 atcaatgcct caggtgtgga tgctaaagct attttatctg ctagattgtc aaaaagtaga
      661 cgtttggaaa atcttattgc ccaattgcca ggtgaaaaaa aaaatggatt gtttggaaac
      721 ttaattgcat tatctttggg tttgacccca aatttcaaat caaactttga tttggccgag
      781 gatgctaaac ttcaattgtc taaggataca tacgatgacg atttagataa tcttcttgcc
      841 caaattggtg atcaatatgc agatttattc ttggcagcta agaacttgag tgatgcaatt
      901 ttgttaagtg acattttgag agtcaacaca gaaattacaa aagctccttt atccgcctct
      961 atgattaaaa gatatgatga acaccatcaa gatttaacct tgttgaaagc tcttgtacgt
     1021 caacaattac cagaaaagta taaagaaata ttctttgatc aatctaaaaa tggttacgct
     1081 ggttatattg atggtggtgc tagtcaagaa gaattttata agtttatcaa gccaattctt
     1141 gaaaaaatgg atggtactga ggaattgttg gttaagttga atcgagagga tttacttcgt
     1201 aagcaaagaa catttgataa cggttctatc cctcatcaaa tccacttagg tgagttacac
     1261 gctatcttga gacgtcagga agatttttac ccttttttga aagacaatcg tgaaaaaatt
     1321 gaaaaaattt tgactttcag aattccttat tacgtcgggc cattggctag aggtaattct
     1381 agatttgcat ggatgactcg taaatctgaa gagactatca ccccatggaa ttttgaggaa
     1441 gttgttgata agggagcctc cgctcaatca ttcatcgaga gaatgaccaa tttcgacaaa
     1501 aacttgccaa atgagaaggt gttacctaaa cattctttgt tatatgaata tttcactgtt
     1561 tataatgaat taacaaaagt caaatacgtc actgaaggta tgagaaaacc agcattctta
     1621 agtggtgaac aaaagaaagc tatcgttgac ttgttattca aaactaatcg taaagtcact
     1681 gtcaaacaac ttaaagagga ctattttaag aaaatagagt gtttcgattc cgttgaaatc
     1741 agtggggttg aggatagatt taatgctagt ttaggaactt atcacgattt gttgaaaatt
     1801 atcaaagaca aagatttttt ggataatgaa gaaaacgaag atatcttgga ggacattgta
     1861 ttgactttga ctttgtttga ggatagagaa atgatcgaag aacgtttaaa aacttacgct
     1921 cacttattcg acgataaagt catgaaacaa ttaaaaagaa gaagatatac cggttgggga
     1981 agattatcta gaaagttgat taatggtatt agagataagc aaagtggtaa aaccatctta
     2041 gatttcctta aaagtgatgg ttttgcaaac agaaacttta tgcagttgat tcatgatgat
     2101 agtttgacct ttaaggaaga tattcaaaaa gctcaggttt ctggacaggg tgattcattg
     2161 catgaacata ttgccaactt agcagggtct ccagctatca aaaagggaat tttacaaact
     2221 gttaaagttg ttgatgagtt ggtgaaagtt atgggacgtc ataagcctga aaatatagtt
     2281 atagaaatgg ctagagaaaa tcaaacaact cagaaaggac aaaagaattc tcgtgaaaga
     2341 atgaaaagaa tagaagaagg aattaaagag ttgggttctc aaatcttaaa agaacaccca
     2401 gtcgaaaaca cccagttaca gaacgagaag ctttacttgt actatttgca aaacgggcgt
     2461 gacatgtatg tagatcaaga gttggatatc aatagacttt cagattacga tgtcgatcac
     2521 atagttcctc agtcattttt aaaagatgat tccattgaca acaaagtttt aacaagatct
     2581 gataagaata gaggtaaatc agacaatgtg ccatcagaag aagtggttaa gaaaatgaag
     2641 aactattgga gacaattgtt aaatgcaaag ttgattactc agagaaaatt tgacaactta
     2701 actaaagctg aaagaggagg attatccgaa ttagataaag ctggattcat aaaacgtcag
     2761 ttggttgaaa caagacaaat tactaaacat gtagcccaaa ttttagattc ccgaatgaat
     2821 actaaatatg atgaaaatga caaattaatc agagaggtca aagtgatcac attaaagtca
     2881 aagttggtgt cagatttccg aaaggatttt caattctaca aggtgagaga gatcaacaat
     2941 tatcatcatg ctcacgacgc atatttgaat gcagtggtag gaactgcctt aataaagaag
     3001 tatcctaagt tagaatccga attcgtgtat ggtgattata aagtctacga tgtcagaaag
     3061 atgatagcaa aatcagaaca agaaattggg aaagccaccg caaagtattt cttttactcc
     3121 aacatcatga atttctttaa gaccgaaatc accttggcta acggggaaat cagaaaacgt
     3181 cctttgattg aaacaaatgg agaaaccggt gaaattgtgt gggataaggg gagagatttt
     3241 gctactgtta gaaaagtttt atcaatgcct caagttaata ttgtaaagaa gactgaagtt
     3301 caaactggtg gtttctctaa ggaatccatt cttccaaaac gaaactcaga caagttgatc
     3361 gcccgaaaaa aagattggga tcctaagaaa tacggaggtt ttgattctcc tactgtcgca
     3421 tattctgtct tggtcgtggc taaagttgaa aagggtaaat ccaaaaaatt gaaatctgtg
     3481 aaagaattgt taggtattac catcatggag cgttcatcct ttgagaaaaa cccaattgac
     3541 tttctcgaag cgaagggcta taaagaagtg aagaaagatt tgattattaa gttgccaaag
     3601 tattctctct ttgaacttga aaatgggaga aagagaatgt tggcttcagc tggagaattg
     3661 caaaaaggaa acgagttagc tttgccttca aagtacgtta atttcttata cttagcatct
     3721 cattatgaaa agttgaaggg gtcccctgaa gacaatgagc aaaagcaatt attcgtagaa
     3781 caacataaac actatttgga tgagataatt gaacaaatct ccgagttttc taaaagagtt
     3841 attttggctg atgcaaattt ggataaagtt ttatctgctt ataataagca tcgtgacaaa
     3901 ccaattcgag aacaagctga aaatattatc cacttgttca cattgactaa cttgggagct
     3961 ccagccgcgt ttaagtactt tgacaccacc attgaccgta agagatatac ttcaacaaaa
     4021 gaagtgttag acgccacatt gattcatcag tccataactg gtttatatga aactagaatc
     4081 gatttgtcac aattgggggg tgatagccga gcggatccta agaaaaaacg aaaagtctga
     4141 ctcgaggtcg acggtatcga taagcttgat atcgaattcc tgcagcccgg gggatccgag
     4201 tgaaattctg gaaatctgga aatctggttt tgtattcttg ttattcttct ttttgttatt
     4261 acatatataa cttgttactt ttttaaaaaa atctttgttt attttataaa tatataaaac
     4321 taaatttaag aaaaagagaa aaatgtttta tttgagagat tgaaatttta cttgaattta
     4381 gcttagcttt tataaagtat tattatgtaa aaaaacaaaa caaatataca ttaaaaagtt
     4441 aagactataa aatagccacc caaggcattt ctatatcttg ttgttgttgt tttcatcttc
     4501 tgtatcagag gaacttattt tattattttc gtcacgggta ttttctcttg tttgatgatt
     4561 catcccattc attccatcat aaaattctag aaggaccacc tttgattgta aatagtaata
     4621 attaccaccc ttatttaatt atttatttaa cttatttatt tatttattat acatatatac
     4681 aaatctaata aagtgaaaat ctcccccttc acatttataa ttggccagtt tttttcaaat
     4741 aagcattcca accagcatct ctataccttt taccttcaat atctggatct cttcctttac
     4801 caaacaatcc tctaccaaca atgataatat cagttccagt gctaacaact tcatcaacag
     4861 ttctatattg ttgtcctaat ccatcacctt tatcatctaa tccaactcca ggtgtcataa
     4921 taagccaatc aaatccttct tcttgtccac ccatatcacg ttgggcaata aatccaataa
     4981 caaattcctt atcggattta gcaatttcaa cagttttttg agaatattct ccatacgaat
     5041 tcgagctcgt ttaaacgtcg acactggatg gcggcgttag tatcgaatcg acagcagtat
     5101 agcgaccagc attcacatac gattgacgca tgatattact ttctgcgcac ttaacttcgc
     5161 atctgggcag atgatgtcga ggcgaaaaaa aatataaatc acgctaacat ttgattaaaa
     5221 tagaacaact acaatataaa aaaactatac aaatgacaag ttcttgaaaa caagaatctt
     5281 tttattgtca gtactgactc gagttattat ggacatggca tagacatata caaagcttgt
     5341 tcaccatcgg aagcagtacc atcgtataaa gcagtatcca aaccacacaa agtgaaaccc
     5401 attcttctat aagcatgaat agctggagca ttaacattgg taacttccaa ccacaaatga
     5461 ccagcacctc tttctctggc gaattcagta gccaaaccca tcaaagctct accaacacca
     5521 tgacctctat gttctggagc aacttcaata tcttcaacag tcaatcttct gttccaacca
     5581 gaataagaaa caacaacgaa accagccaaa tcaccatcat caccataagc aacgaaagtt
     5641 ctagaatctg gatcaccatc ttcaccagca tcggattcat catcggattc atcatctggg
     5701 aaaaccttag tcaatggtgg atcaactgga acttctctca aagtgaaacc atcaccagta
     5761 gcagtaactc taaaaacagt atcggtagtg aaagaaccat ccaaagcttc aatagcttca
     5821 gcatcacctg gaacagaagt tctgtatcta taagcagtat catccaaagt agtagacata
     5881 attgtaggat ccggttgttt atgttcggat gtgatgtgag aactgtatcc tagcaagatt
     5941 ttaaaaggaa gtatatgaaa gaagaacctc agtggcaaat cctaaccttt tatatttctc
     6001 tacaggggcg cggcgtgggg acaattcaac gcgtctgtga ggggagcgtt tccctgctcg
     6061 caggtctgca gcgaggagcc gtaatttttg cttcgcgccg tgcggccatc aaaatgtatg
     6121 gatgcaaatg attatacatg gggatgtatg ggctaaatgt acgggcgaca gtcacatcat
     6181 gcccctgagc tgcgcacgtc aagactgtca aggagggtat tctgggcctc catgtcgctg
     6241 gccgggtgac ccggcgggga cgaggcaagc ttgatatgct aatgatccca ctgatgataa
     6301 ttcagctaac atcaataacc ctcttggctc ttggttggtg gtggtttctt tagctccctg
     6361 ttttaatcct tcaactactc cattcccagt gacaccatga gcattagtaa tatctgccca
     6421 actactaatt ttataaactc caccaatata ttgtttcttc acggtattac caatatcagc
     6481 aaattttcta tcttcaaaaa tcataaattg atgtttacgt gaaagttcta ataatggttc
     6541 aatagtggat tcataggaaa aatcattgat tatatcaata tgagtcttga ttaagcatac
     6601 ataaggaccc aatttatcaa ttaattcaag gaattcctta gtggtatcaa catcaattga
     6661 tgcacataaa ttggttttct tcagttccat taatcgaaat aatcgttgtg ctactggtga
     6721 ggcatgagtt tctgctctct cactataggt cttagtgttg actgtcatat cttttgtttg
     6781 ctatttctat ttctatttct ttcttcaact gtaaaagggg cgatatataa accagtgtgt
     6841 atggggttgt tgctcaggaa ttgatcaatg tttaaaagaa tggttgaaat catcattgaa
     6901 gaacaaaaaa aaagattttt tctttcatta tgttatctgt ataatatgca ggaaataaga
     6961 ttgcaaaaaa atagccgttt ttttggatac tatcaaacaa gaggaatgac taatacagaa
     7021 ttagtgttac gaatcaatgg cactacagca actttcaaca aaataatcta cgttcaaaat
     7081 ataatatata ctattaaata aaatcaaata ataaaataat tttaaagaaa gtagtaccag
     7141 ctcttttttt tgtttccgtt tataccatcc aaatcaattc ctattagtct ccaattcgcc
     7201 ctatagtgag tcgtattaca attcactggc cgtcgtttta caacgtcgtg actgggaaaa
     7261 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     7321 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     7381 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     7441 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     7501 cacgttcgcc tttaaatgat aattatagta ttcacggata gatctgttag agatgtggtg
     7561 gattggtatt tctttctgtg attcaccgaa agtattatca actaagaagg gaaaggcacc
     7621 acattttcta atgtctcata tattagttca gttcgatgta tttcttttta tatattaatt
     7681 ataattttat tgcaccataa cgtttacttg tttaatatgc tattgatatc tatatgcggc
     7741 cagacataaa aaacaaaaaa agcaccaccg actcggtgcc actttttcaa gttgataacg
     7801 gactagcctt attttaactt gctatttcta gctctaaaac attaggtcgt atgattgttg
     7861 cgcaccaccg actcggtgcc actttttcaa gttgataacg gactagcctt attttaactt
     7921 gctatttcta gctctaaaac atttcttcta aagtttctgc caaattaaaa atagtttacg
     7981 caagtctcga acttccacca caatttaatt gtaagaacgc tgcgaaaaca tattctccaa
     8041 tcctattggg ccgaagtttc agctatgcca aaggcatcaa attttttttt ataaaatttt
     8101 ccacttttcc ataccagtga aattatcacg tgatggagtg agtatctata atatttgatt
     8161 gtttctttga tgaggcaata ctcgagtata gttattgacc aatgactttt agctatgtgt
     8221 agtgcttgta tatgcgtgtg tgatgagaat aagatgaata gattagtaat gtataagttg
     8281 attctttttg gtggggggtg ttacttttat tttttctttc catctcattg acaagattgt
     8341 accgagtcaa aatgtttggg gtctttttat tgctgcgatg gattgatttt tcactctaag
     8401 cctaaaatta aatggctaaa caagtggtat tcaagcacaa ttcagttttc tggatctcta
     8461 ataattgcaa ttgcaattgc ttcacatact ctgtttataa ttcagtttct ttctcttgat
     8521 gttactgata tattaagtta ataaatacca ataacttttt ttcaaaaatc ttctattttc
     8581 ctttcaattg attatttaaa cacgaatcgt taataagctg tgattgcttt ttctctttct
     8641 ttttcttttc tttgttcaag tttgaaaaaa aaaaaaaaag gtttctcttg ttcgatgtag
     8701 aattcagtgg gttatagttg taatgatata tggaataaaa agaaagaaag agaagagaaa
     8761 gagaagaaga gaagaattaa gtaaacgtgt attagaattt aaaaattaaa attcatgata
     8821 atctttaata ataattaact aagaatcaca aaattgaaaa tttgaaagaa aaaagatcac
     8881 tccaaggtcg tgtcaagtga ctagaaattc ttaattaaaa tagttataac aaaaaaagaa
     8941 atttgtatat aacaattaag taaaagaaaa aaaagcaatt cgctttgtat aataataccc
     9001 agaaaaatat attcatatat tcataaatca aacatgaaat gaagaatgct gaatcacttg
     9061 ataggattta gttccattat ggtactaaga agtaatatta cttctatgaa aaggttgaat
     9121 ccgagatcaa aattcatctt ttcatatact ctggagaaaa aaaaaaaagt aattgtagta
     9181 agaatgacaa gtatcagttg agaaaaaaag taacaaagag aaagctcgga ggaggctccc
     9241 caaaggtttt atcaccaatg gtggtaccac taacccaaga acagaaaaag cattgatatt
     9301 ttgaaaaata caatactcat cttttaaccc aaaccttttc ttgtccttta aaacttttat
     9361 ctcatgtctg tccaaagtca tattgtctta ctactttctc tccttcaacg tccttccggc
     9421 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     9481 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     9541 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     9601 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     9661 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     9721 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc
     9781 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     9841 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     9901 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     9961 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
    10021 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
    10081 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
    10141 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
    10201 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
    10261 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
    10321 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
    10381 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
    10441 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
    10501 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
    10561 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
    10621 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
    10681 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
    10741 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
    10801 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
    10861 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
    10921 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
    10981 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
    11041 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
    11101 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
    11161 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
    11221 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
    11281 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
    11341 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
    11401 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
    11461 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
    11521 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
    11581 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
    11641 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
    11701 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
    11761 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
    11821 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
    11881 caggaaacag ctatgaccat gattacgcca agctcgaaat taaccctcac taaagggaac
    11941 aaaagctggt accagagcta ttaagatcac cagcctcgtt tataataaac ttagtctaat
    12001 atttcttttg gaccaaaata gaaactgttc tcaggtggca aaaaacagct taaagtctgt
    12061 gttaaaaaaa gatctcggct atgccaatca aaaaggagtg acattattat gtaaaagaag
    12121 aaatgttgta attccgcgaa acaacaattt taatgcttac tgtgccgcca ccaatatcct
    12181 atggccaagg ggcctttaaa catgccacgg aaatatacag tcaaagtaaa cctactatgt
    12241 ctttagagcc ttcaggatca attgttttta aatgaatgac tgcaaaacta gaaaagtagc
    12301 ctcttacata agcaagtcat ttgtacaccg agagtttagt aaccagtgaa gcaagtatat
    12361 ctggcaaacc tgaaacttat ctgtgaatta taaagctcat aagtaaaaag tgtagaaaag
    12421 aagataaggc agaccaaata caaatttgca acagtacaac gaatatttca gcgtcaaaac
    12481 tagagaataa taaagaaaac gatcttttca aaaagaaaaa accttttagt tttcctttgt
    12541 tgttgttgtg ggtgtgtgct atttatatta tatagtttac tcataatacc ataaaatatt
    12601 cggtttgatt aggttatttt aataagctaa tttgtttcta atcgtgtaat ttatgctgtg
    12661 tatattaagt agtgtgtgca ctgcccaaaa atgtttgttg tttatagtcg gttaaagaga
    12721 aaaaagaaaa aaagatccat acacacacgt taattagttg ttcaacgtaa tacactcata
    12781 ttttgttctt atttgctttc ggtcgctgtt ctcaccaaga tttattgcca acgaaacaat
    12841 ttttttttat atattttcag atttttcttt ttttcctttc ctttcctttt ctaattttca
    12901 ctcctggttt tctttctttc ttagaaacat tatctcgata ttaatattaa aaaaatataa
    12961 tcattcaaa
//

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