p201N Cas9

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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p201NCas9载体基本信息

质粒名称: p201N Cas9 
出品公司: 爱迪基因
目录编号: 59175
存储实验室: Wayne Parrott

载体骨架基本信息
载体名称: pPZP
载体抗性: 卡那霉素
空载体大小: 6200 bp
完整质粒大小: 14349 bp
载体修饰: 利用aph基因对卡那霉素进行细菌筛选。
载体类型: CRISPR
筛选标记: G418,卡那霉素
高拷贝/低拷贝: 高拷贝
生长条件: 37 ℃

插入基因信息
插入基因名称: Cas9,Cas9 human-codon optimized
物种来源: Synthetic
插入大小: 5265 bp
启动子 2x35S
克隆方法: 限制性内切酶
5' 克隆位点: I-SceI(未销毁)
3' 克隆位点: I-SceI(未销毁)
5' 测序引物 ACATGCACCTAATTTCACTAGATGT
3' 测序引物 TGTAAAACGACGGCCAGT

文献引用方法
材料和方法部分: p201N Cas9 was a gift from Wayne Parrott (Addgene plasmid # 59175 ; http://n2t.net/addgene:59175 ; RRID:Addgene_59175)
参考文献部分: Targeted genome modifications in soybean with CRISPR/Cas9. Jacobs TB, LaFayette PR, Schmitz RJ, Parrott WA. . BMC Biotechnology. 2015;15:16 10.1186/s12896-015-0131-2

p201NCas9载体图谱质粒图谱和多克隆位点信息

p201N Cas9质粒图谱

p201NCas9载体简介

p201NCas9载体序列

LOCUS       p201N Cas9              14414 bp ds-DNA     circular SYN 18-DEC-2019
DEFINITION  Cas9 driven by double 35S, nptII for plant selection, I-PpoI site to
            accept gRNA from pUC gRNA Shuttle.
ACCESSION   .
VERSION     .
KEYWORDS    p201N Cas9
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14414)
  AUTHORS   Jacobs TB, LaFayette PR, Schmitz RJ, Parrott WA.
  TITLE     Targeted genome modifications in soybean with CRISPR/Cas9.
  JOURNAL   BMC Biotechnology. 2015;15:16
REFERENCE   2  (bases 1 to 14414)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 18, 2019 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..14414
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             1217..1846
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             2275..3348
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      3414..3608
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(3844..3866)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3952..4092
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4007..4026
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4107..4124)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4278..4866)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4358..4377)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4961..5776)
                     /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     5684..5703
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5768..5787
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     misc_feature    6069..6093
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     protein_bind    6270..6291
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6306..6336
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6344..6360
                     /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     6349..6371
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6368..6384
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6368..6384
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(6906..7700)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7017..7036)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     7627..7646
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        8846..9522
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     9459..9479
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             9568..13671
                     /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             13684..13704
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      13771..14023
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(14064..14081)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(14064..14080)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14073..14095)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    14276..14300
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
ORIGIN
        1 agtactttaa agtactttaa agtactttaa agtactttga tccaacccct ccgctgctat
       61 agtgcagtcg gcttctgacg ttcagtgcag ccgtcttctg aaaacgacat gtcgcacaag
      121 tcctaagtta cgcgacaggc tgccgccctg cccttttcct ggcgttttct tgtcgcgtgt
      181 tttagtcgca taaagtagaa tacttgcgac tagaaccgga gacattacgc catgaacaag
      241 agcgccgccg ctggcctgct gggctatgcc cgcgtcagca ccgacgacca ggacttgacc
      301 aaccaacggg ccgaactgca cgcggccggc tgcaccaagc tgttttccga gaagatcacc
      361 ggcaccaggc gcgaccgccc ggagctggcc aggatgcttg accacctacg ccctggcgac
      421 gttgtgacag tgaccaggct agaccgcctg gcccgcagca cccgcgacct actggacatt
      481 gccgagcgca tccaggaggc cggcgcgggc ctgcgtagcc tggcagagcc gtgggccgac
      541 accaccacgc cggccggccg catggtgttg accgtgttcg ccggcattgc cgagttcgag
      601 cgttccctaa tcatcgaccg cacccggagc gggcgcgagg ccgccaaggc ccgaggcgtg
      661 aagtttggcc cccgccctac cctcaccccg gcacagatcg cgcacgcccg cgagctgatc
      721 gaccaggaag gccgcaccgt gaaagaggcg gctgcactgc ttggcgtgca tcgctcgacc
      781 ctgtaccgcg cacttgagcg cagcgaggaa gtgacgccca ccgaggccag gcggcgcggt
      841 gccttccgtg aggacgcatt gaccgaggcc gacgccctgg cggccgccga gaatgaacgc
      901 caagaggaac aagcatgaaa ccgcaccagg acggccagga cgaaccgttt ttcattaccg
      961 aagagatcga ggcggagatg atcgcggccg ggtacgtgtt cgagccgccc gcgcacgtct
     1021 caaccgtgcg gctgcatgaa atcctggccg gtttgtctga tgccaagctg gcggcctggc
     1081 cggccagctt ggccgctgaa gaaaccgagc gccgccgtct aaaaaggtga tgtgtatttg
     1141 agtaaaacag cttgcgtcat gcggtcgctg cgtatatgat gcgatgagta aataaacaaa
     1201 tacgcaaggg gaacgcatga aggttatcgc tgtacttaac cagaaaggcg ggtcaggcaa
     1261 gacgaccatc gcaacccatc tagcccgcgc cctgcaactc gccggggccg atgttctgtt
     1321 agtcgattcc gatccccagg gcagtgcccg cgattgggcg gccgtgcggg aagatcaacc
     1381 gctaaccgtt gtcggcatcg accgcccgac gattgaccgc gacgtgaagg ccatcggccg
     1441 gcgcgacttc gtagtgatcg acggagcgcc ccaggcggcg gacttggctg tgtccgcgat
     1501 caaggcagcc gacttcgtgc tgattccggt gcagccaagc ccttacgaca tatgggccac
     1561 cgccgacctg gtggagctgg ttaagcagcg cattgaggtc acggatggaa ggctacaagc
     1621 ggcctttgtc gtgtcgcggg cgatcaaagg cacgcgcatc ggcggtgagg ttgccgaggc
     1681 gctggccggg tacgagctgc ccattcttga gtcccgtatc acgcagcgcg tgagctaccc
     1741 aggcactgcc gccgccggca caaccgttct tgaatcagaa cccgagggcg acgctgcccg
     1801 cgaggtccag gcgctggccg ctgaaattaa atcaaaactc atttgagtta atgaggtaaa
     1861 gagaaaatga gcaaaagcac aaacacgcta agtgccggcc gtccgagcgc acgcagcagc
     1921 aaggctgcaa cgttggccag cctggcagac acgccagcca tgaagcgggt caactttcag
     1981 ttgccggcgg aggatcacac caagctgaag atgtacgcgg tacgccaagg caagaccatt
     2041 accgagctgc tatctgaata catcgcgcag ctaccagagt aaatgagcaa atgaataaat
     2101 gagtagatga attttagcgg ctaaaggagg cggcatggaa aatcaagaac aaccaggcac
     2161 cgacgccgtg gaatgcccca tgtgtggagg aacgggcggt tggccaggcg taagcggctg
     2221 ggttgtctgc cggccctgca atggcactgg aacccccaag cccgaggaat cggcgtgagc
     2281 ggtcgcaaac catccggccc ggtacaaatc ggcgcggcgc tgggtgatga cctggtggag
     2341 aagttgaagg ccgcgcaggc cgcccagcgg caacgcatcg aggcagaagc acgccccggt
     2401 gaatcgtggc aagcggccgc tgatcgaatc cgcaaagaat cccggcaacc gccggcagcc
     2461 ggtgcgccgt cgattaggaa gccgcccaag ggcgacgagc aaccagattt tttcgttccg
     2521 atgctctatg acgtgggcac ccgcgatagt cgcagcatca tggacgtggc cgttttccgt
     2581 ctgtcgaagc gtgaccgacg agctggcgag gtgatccgct acgagcttcc agacgggcac
     2641 gtagaggttt ccgcagggcc ggccggcatg gccagtgtgt gggattacga cctggtactg
     2701 atggcggttt cccatctaac cgaatccatg aaccgatacc gggaagggaa gggagacaag
     2761 cccggccgcg tgttccgtcc acacgttgcg gacgtactca agttctgccg gcgagccgat
     2821 ggcggaaagc agaaagacga cctggtagaa acctgcattc ggttaaacac cacgcacgtt
     2881 gccatgcagc gtacgaagaa ggccaagaac ggccgcctgg tgacggtatc cgagggtgaa
     2941 gccttgatta gccgctacaa gatcgtaaag agcgaaaccg ggcggccgga gtacatcgag
     3001 atcgagctag ctgattggat gtaccgcgag atcacagaag gcaagaaccc ggacgtgctg
     3061 acggttcacc ccgattactt tttgatcgat cccggcatcg gccgttttct ctaccgcctg
     3121 gcacgccgcg ccgcaggcaa ggcagaagcc agatggttgt tcaagacgat ctacgaacgc
     3181 agtggcagcg ccggagagtt caagaagttc tgtttcaccg tgcgcaagct gatcgggtca
     3241 aatgacctgc cggagtacga tttgaaggag gaggcggggc aggctggccc gatcctagtc
     3301 atgcgctacc gcaacctgat cgagggcgaa gcatccgccg gttcctaatg tacggagcag
     3361 atgctagggc aaattgccct agcaggggaa aaaggtcgaa aaggtctctt tcctgtggat
     3421 agcacgtaca ttgggaaccc aaagccgtac attgggaacc ggaacccgta cattgggaac
     3481 ccaaagccgt acattgggaa ccggtcacac atgtaagtga ctgatataaa agagaaaaaa
     3541 ggcgattttt ccgcctaaaa ctctttaaaa cttattaaaa ctcttaaaac ccgcctggcc
     3601 tgtgcataac tgtctggcca gcgcacagcc gaagagctgc aaaaagcgcc tacccttcgg
     3661 tcgctgcgct ccctacgccc cgccgcttcg cgtcggccta tcgcggccgc tggccgctca
     3721 aaaatggctg gcctacggcc aggcaatcta ccagggcgcg gacaagccgc gccgtcgcca
     3781 ctcgaccgcc ggcgcccaca tcaaggcacc ctgcctcgcg cgtttcggtg atgacggtga
     3841 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     3901 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat
     3961 gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag
     4021 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     4081 taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4141 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4201 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4261 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4321 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4381 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4441 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4501 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4561 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4621 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4681 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     4741 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4801 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4861 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4921 cgttaaggga ttttggtcat gcaggatcat gaattaattc ttagaaaaac tcatcgagca
     4981 tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc
     5041 gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt
     5101 atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa
     5161 aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca
     5221 aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa
     5281 aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata
     5341 cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca
     5401 ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg
     5461 ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat
     5521 gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg
     5581 taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct
     5641 tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat
     5701 acccatataa atcagcatcc atgttggaat ttaatcgcgg cctagagcaa gacgtttccc
     5761 gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg
     5821 ttcatgatct ggatcacagg cagcaacgct ctgtcatcgt tacaatcaac atgctaccct
     5881 ccgcgagatc atccgtgttt caaacccggc agcttagttg ccgttcttcc gaatagcatc
     5941 ggtaacatga gcaaagtctg ccgccttaca acggctctcc cgctgacgcc gtcccggact
     6001 gatgggctgc ctgtatcgag tggtgatttt gtgccgagct gccggtcggg gagctgttgg
     6061 ctggctggtg gcaggatata ttgtggtgta aacaaattga cgcttagaca acttaataac
     6121 acattgcgga cgtttttaat gtactgaatt aacgccgaat tgctctagcc aatacgcaaa
     6181 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     6241 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     6301 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     6361 tttcacacag gaaacagcta tgacatgatt acgaatcctt gactagaggg taggcgcggc
     6421 ttaattaagc gcttaattaa gcgcttaatt aagcgcttaa taagccgcgc cgtaccccaa
     6481 tagtctcgac agacacatag cacctaacat tattgcttaa atttactatt tttttactat
     6541 attatactca acccaatgag cataaagact gtaaaatctc aaattcctga gaagcatatt
     6601 tatcgatccc acagacttga tagttccata atccatacgc tgcagccaaa ttgctagtgt
     6661 gttgaacatt taacacgtag agaactagaa aagatataaa actaagattg atatccaaaa
     6721 tagacgagaa caataagcaa aaactcttag ttttgaaata aatcaacaat cccgagggtt
     6781 gtcacataca tcaaaaacga aaatccatat agcaaaaaaa actctaaatt accgttcgac
     6841 aaaaagagaa aactgatagg acatttgcta aacattaaaa tcaaaatttg gatcaagccg
     6901 aattctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg
     6961 gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata
     7021 tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg
     7081 atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg
     7141 gtcacgacga gatcatcgcc gtcgggcatg cgcgccttga gcctggcgaa cagttcggct
     7201 ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc
     7261 cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga
     7321 tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca
     7381 aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc
     7441 gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat
     7501 agccgcgctg cctcgtcctg cagttcattc agggcaccgg acaggtcggt cttgacaaaa
     7561 agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc
     7621 tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc
     7681 aatccatctt gttcaatcat ctcgagacgt accgcgcgat gcattcgaag atcgatcctt
     7741 cgcctggagg agagaaatca gtggcgctgc ggcttttagg gtttctttgt tgatggaatg
     7801 agagtgtaag ctctgccagt gccactttat tagggtttta caagcccttt tcttcgtaat
     7861 tgggcctgac attttgtgcc acttgggcct ttagagatga aaatgtatat tgggcttaag
     7921 ttgacttgaa ggataaatta gtttaggata ttacgttttt tatgagaatt ggtgtgtcgg
     7981 atacatgtat atgatgcatt caaatatatg tattctagat acatttaagt ttagatacaa
     8041 tctaaaatgt gtctttaatt acaggactgt aactaaaata cttaatgtaa gaagaatatt
     8101 actcctttaa tagcttttga gtatatctag tctaacatct tttaaaaaag tctaatttct
     8161 ttcatttatt tttcgagcaa tagcaaagtg cataattatt tttttcttct agaaattcag
     8221 atttgtttct ctaaattttg agattctttt ctcaattttg tatgtctaga gaacaatgtg
     8281 tatttttcac tctagttggt tgttactttg ttgaatgttc tgataaaagt atattgttat
     8341 ttctgaagta gatataaacc ttcatttgga aattatacat aaatcaaaat cgttaattat
     8401 ctagatcaag atatatgccc ttttcctaat gtatttgata catgcaccta atttcactag
     8461 atgtatcttt tctatttttt aaattatgaa tagttaattt tttccatatg tgtatttgat
     8521 acatacttca tgactttaaa aaattaatta tataccagat atatgtattt aaaatttgtt
     8581 atgtatttaa agtatgtata tgattattcg atattaatct cttcgatgaa atttaaatcg
     8641 ataactatgt gctttggatc gatctgccca ctagtgagtc gtattacaat tcatgactct
     8701 cttaaggtag ccaaaatcga tatctaggga taacagggta atcgatcact agaattcggc
     8761 gcgccgggcc caacatggtg gagcacgaca ctctcgtcta ctccaagaat atcaaagata
     8821 cagtctcaga agaccaaagg gctattgaga cttttcaaca aagggtaata tcgggaaacc
     8881 tcctcggatt ccattgccca gctatctgtc acttcatcaa aaggacagta gaaaaggaag
     8941 gtggcaccta caaatgccat cattgcgata aaggaaaggc tatcgttcaa gatgcctctg
     9001 ccgacagtgg tcccaaagat ggacccccac ccacgaggag catcgtggaa aaagaagacg
     9061 ttccaaccac gtcttcaaag caagtggatt gatgtgaaca tggtggagca cgacactctc
     9121 gtctactcca agaatatcaa agatacagtc tcagaagacc aaagggctat tgagactttt
     9181 caacaaaggg taatatcggg aaacctcctc ggattccatt gcccagctat ctgtcacttc
     9241 atcaaaagga cagtagaaaa ggaaggtggc acctacaaat gccatcattg cgataaagga
     9301 aaggctatcg ttcaagatgc ctctgccgac agtggtccca aagatggacc cccacccacg
     9361 aggagcatcg tggaaaaaga agacgttcca accacgtctt caaagcaagt ggattgatgt
     9421 gatatctcca ctgacgtaag ggatgacgca caatcccact atccttcgca agacccttcc
     9481 tctatataag gaagttcatt tcatttggag aggacacgct gaaatcacca gtctctctct
     9541 acaaatctat ctctctcgac cgctagcatg gacaagaagt actccattgg gctcgatatc
     9601 ggcacaaaca gcgtcggctg ggccgtcatt acggacgagt acaaggtgcc gagcaaaaaa
     9661 ttcaaagttc tgggcaatac cgatcgccac agcataaaga agaacctcat tggcgccctc
     9721 ctgttcgact ccggggagac ggccgaagcc acgcggctca aaagaacagc acggcgcaga
     9781 tatacccgca gaaagaatcg gatctgctac ctgcaggaga tctttagtaa tgagatggct
     9841 aaggtggatg actctttctt ccataggctg gaggagtcct ttttggtgga ggaggataaa
     9901 aagcacgagc gccacccaat ctttggcaat atcgtggacg aggtggcgta ccatgaaaag
     9961 tacccaacca tatatcatct gaggaagaag cttgtagaca gtactgataa ggctgacttg
    10021 cggttgatct atctcgcgct ggcgcatatg atcaaatttc ggggacactt cctcatcgag
    10081 ggggacctga acccagacaa cagcgatgtc gacaaactct ttatccaact ggttcagact
    10141 tacaatcagc ttttcgaaga gaacccgatc aacgcatccg gagttgacgc caaagcaatc
    10201 ctgagcgcta ggctgtccaa atcccggcgg ctcgaaaacc tcatcgcaca gctccctggg
    10261 gagaagaaga acggcctgtt tggtaatctt atcgccctgt cactcgggct gacccccaac
    10321 tttaaatcta acttcgacct ggccgaagat gccaagcttc aactgagcaa agacacctac
    10381 gatgatgatc tcgacaatct gctggcccag atcggcgacc agtacgcaga cctttttttg
    10441 gcggcaaaga acctgtcaga cgccattctg ctgagtgata ttctgcgagt gaacacggag
    10501 atcaccaaag ctccgctgag cgctagtatg atcaagcgct atgatgagca ccaccaagac
    10561 ttgactttgc tgaaggccct tgtcagacag caactgcctg agaagtacaa ggaaattttc
    10621 ttcgatcagt ctaaaaatgg ctacgccgga tacattgacg gcggagcaag ccaggaggaa
    10681 ttttacaaat ttattaagcc catcttggaa aaaatggacg gcaccgagga gctgctggta
    10741 aagcttaaca gagaagatct gttgcgcaaa cagcgcactt tcgacaatgg aagcatcccc
    10801 caccagattc acctgggcga actgcacgct atcctcaggc ggcaagagga tttctacccc
    10861 tttttgaaag ataacaggga aaagattgag aaaatcctca catttcggat accctactat
    10921 gtaggccccc tcgcccgggg aaattccaga ttcgcgtgga tgactcgcaa atcagaagag
    10981 accatcactc cctggaactt cgaggaagtc gtggataagg gggcctctgc ccagtccttc
    11041 atcgaaagga tgactaactt tgataaaaat ctgcctaacg aaaaggtgct tcctaaacac
    11101 tctctgctgt acgagtactt cacagtttat aacgagctca ccaaggtcaa atacgtcaca
    11161 gaagggatga gaaagccagc attcctgtct ggagagcaga agaaagctat cgtggacctc
    11221 ctcttcaaga cgaaccggaa agttaccgtg aaacagctca aagaagacta tttcaaaaag
    11281 attgaatgtt tcgactctgt tgaaatcagc ggagtggagg atcgcttcaa cgcatccctg
    11341 ggaacgtatc acgatctcct gaaaatcatt aaagacaagg acttcctgga caatgaggag
    11401 aacgaggaca ttcttgagga cattgtcctc acccttacgt tgtttgaaga tagggagatg
    11461 attgaagaac gcttgaaaac ttacgctcat ctcttcgacg acaaagtcat gaaacagctc
    11521 aagaggcgcc gatatacagg atgggggcgg ctgtcaagaa aactgatcaa tgggatccga
    11581 gacaagcaga gtggaaagac aatcctggat tttcttaagt ccgatggatt tgccaaccgg
    11641 aacttcatgc agttgatcca tgatgactct ctcaccttta aggaggacat ccagaaagca
    11701 caagtttctg gccaggggga cagtcttcac gagcacatcg ctaatcttgc aggtagccca
    11761 gctatcaaaa agggaatact gcagaccgtt aaggtcgtgg atgaactcgt caaagtaatg
    11821 ggaaggcata agcccgagaa tatcgttatc gagatggccc gagagaacca aactacccag
    11881 aagggacaga agaacagtag ggaaaggatg aagaggattg aagagggtat aaaagaactg
    11941 gggtcccaaa tccttaagga acacccagtt gaaaacaccc agcttcagaa tgagaagctc
    12001 tacctgtact acctgcagaa cggcagggac atgtacgtgg atcaggaact ggacatcaat
    12061 cggctctccg actacgacgt ggatcatatc gtgccccagt cttttctcaa agatgattct
    12121 attgataata aagtgttgac aagatccgat aaaaatagag ggaagagtga taacgtcccc
    12181 tcagaagaag ttgtcaagaa aatgaaaaat tattggcggc agctgctgaa cgccaaactg
    12241 atcacacaac ggaagttcga taatctgact aaggctgaac gaggtggcct gtctgagttg
    12301 gataaagccg gcttcatcaa aaggcagctt gttgagacac gccagatcac caagcacgtg
    12361 gcccaaattc tcgattcacg catgaacacc aagtacgatg aaaatgacaa actgattcga
    12421 gaggtgaaag ttattactct gaagtctaag ctggtctcag atttcagaaa ggactttcag
    12481 ttttataagg tgagagagat caacaattac caccatgcgc atgatgccta cctgaatgca
    12541 gtggtaggca ctgcacttat caaaaaatat cccaagcttg aatctgaatt tgtttacgga
    12601 gactataaag tgtacgatgt taggaaaatg atcgcaaagt ctgagcagga aataggcaag
    12661 gccaccgcta agtacttctt ttacagcaat attatgaatt ttttcaagac cgagattaca
    12721 ctggccaatg gagagattcg gaagcgacca cttatcgaaa caaacggaga aacaggagaa
    12781 atcgtgtggg acaagggtag ggatttcgcg acagtccgga aggtcctgtc catgccgcag
    12841 gtgaacatcg ttaaaaagac cgaagtacag accggaggct tctccaagga aagtatcctc
    12901 ccgaaaagga acagcgacaa gctgatcgca cgcaaaaaag attgggaccc caagaaatac
    12961 ggcggattcg attctcctac agtcgcttac agtgtactgg ttgtggccaa agtggagaaa
    13021 gggaagtcta aaaaactcaa aagcgtcaag gaactgctgg gcatcacaat catggagcga
    13081 tcaagcttcg aaaaaaaccc catcgacttt ctcgaggcga aaggatataa agaggtcaaa
    13141 aaagacctca tcattaagct tcccaagtac tctctctttg agcttgaaaa cggccggaaa
    13201 cgaatgctcg ctagtgcggg cgagctgcag aaaggtaacg agctggcact gccctctaaa
    13261 tacgttaatt tcttgtatct ggccagccac tatgaaaagc tcaaagggtc tcccgaagat
    13321 aatgagcaga agcagctgtt cgtggaacaa cacaaacact accttgatga gatcatcgag
    13381 caaataagcg aattctccaa aagagtgatc ctcgccgacg ctaacctcga taaggtgctt
    13441 tctgcttaca ataagcacag ggataagccc atcagggagc aggcagaaaa cattatccac
    13501 ttgtttactc tgaccaactt gggcgcgcct gcagccttca agtacttcga caccaccata
    13561 gacagaaagc ggtacacctc tacaaaggag gtcctggacg ccacactgat tcatcagtca
    13621 attacggggc tctatgaaac aagaatcgac ctctctcagc tcggtggaga cagcagggct
    13681 gaccccaaga agaagaggaa ggtgtgaccg cggatatctg cagaagctta cgcgtcgacg
    13741 tctcgaggcc ggccgagctc gaatttcccc gatcgttcaa acatttggca ataaagtttc
    13801 ttaagattga atcctgttgc cggtcttgcg atgattatca tataatttct gttgaattac
    13861 gttaagcatg taataattaa catgtaatgc atgacgttat ttatgagatg ggtttttatg
    13921 attagagtcc cgcaattata catttaatac gcgatagaaa acaaaatata gcgcgcaaac
    13981 taggataaat tatcgcgcgc ggtgtcatct atgttactag atcgggaatt gatatctagg
    14041 gataacaggg taataagctt ggcactggcc gtcgttttac aacgtcgtga ctgggaaaac
    14101 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
    14161 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatga
    14221 gcttgagctt ggatcagatt gtcgtttccc gccttcagtt taaactatca gtgtttgaca
    14281 ggatatattg gcgggtaaac ctaagagaaa agagcgttta ttagaataat cggatattta
    14341 aaagggcgtg aaaaggttta tccgttcgtc catttgtatg tgcatgccaa ccacagggtt
    14401 cccctcggga tcaa
//

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