pUB-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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pUBCas9载体基本信息

质粒名称: pUB-Cas9 
出品公司: 爱迪基因
目录编号: 86556
存储实验室: Andreas Weber

载体骨架基本信息
载体名称: pKB65
载体出品公司: Kristin Bernhardt
载体抗性: 卡那霉素
空载体大小: 11630 bp
完整质粒大小: 14121 bp
载体修饰: --
载体类型: 植物表达,CRISPR
筛选标记: 潮霉素
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
物种来源: Synthetic
插入大小: 4137 bp
基因突变: Cas9 is codon optimized for C. reinhardtii
GenBank编号 --
启动子 UBIQUITIN10
标签/融合蛋白: --
克隆方法: Gateway克隆
5' 克隆位点: --
3' 克隆位点: --
5' 测序引物 TTTCGTTCGATCCCAATTTC
3' 测序引物 aagaccggcaacaggattc

文献引用方法
材料和方法部分: pUB-Cas9 was a gift from Andreas Weber (Addgene plasmid # 86556 ; http://n2t.net/addgene:86556 ; RRID:Addgene_86556)
参考文献部分: An Efficient Visual Screen for CRISPR/Cas9 Activity in Arabidopsis thaliana. Hahn F, Mantegazza O, Greiner A, Hegemann P, Eisenhut M, Weber AP. Front Plant Sci. 2017 Jan 24;8:39. doi: 10.3389/fpls.2017.00039. eCollection 2017. 10.3389/fpls.2017.00039 PubMed 28174584

pUBCas9载体图谱质粒图谱和多克隆位点信息

pUB-Cas9质粒图谱

pUBCas9载体简介

pUBCas9载体序列

LOCUS       pUB-Cas9              14128 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  For ubiquitous expression of Cas9 in planta. sgRNA can be introduced
            as described in Hahn et al. (2017).
ACCESSION   .
VERSION     .
KEYWORDS    pUB-Cas9
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14128)
  AUTHORS   Hahn F, Mantegazza O, Greiner A, Hegemann P, Eisenhut M, Weber AP
  TITLE     An Efficient Visual Screen for CRISPR/Cas9 Activity in Arabidopsis 
            thaliana.
  JOURNAL   Front Plant Sci. 2017 Jan 24;8:39. doi: 10.3389/fpls.2017.00039. 
            eCollection 2017.
  PUBMED    28174584
REFERENCE   2  (bases 1 to 14128)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..14128
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(12..28)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12..28)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(25..47)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    36..52
                     /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)."
     promoter        complement(60..90)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    105..126
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        317..993
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     930..950
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             1060..2085
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     polyA_signal    2126..2300
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    2378..2402
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             2827..3621
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      3708..4296
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4197..4216
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4450..4467
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4482..4622
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4548..4567)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4708..4730
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      4966..5160
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(5226..6299)
                     /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"
     CDS             complement(6728..7357)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    8658..8682
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     8870..8892
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8884..8901
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8885..8901
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    9610..9634
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             9655..9675
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9676..13779
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     protein_bind    complement(13783..13807)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      13871..14123
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
ORIGIN
        1 ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
       61 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
      121 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
      181 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattggcta gagcagcttg
      241 ccaacatggt ggagcacgac actctcgtct actccaagaa tatcaaagat acagtctcag
      301 aagaccaaag ggctattgag acttttcaac aaagggtaat atcgggaaac ctcctcggat
      361 tccattgccc agctatctgt cacttcatca aaaggacagt agaaaaggaa ggtggcacct
      421 acaaatgcca tcattgcgat aaaggaaagg ctatcgttca agatgcctct gccgacagtg
      481 gtcccaaaga tggaccccca cccacgagga gcatcgtgga aaaagaagac gttccaacca
      541 cgtcttcaaa gcaagtggat tgatgtgaac atggtggagc acgacactct cgtctactcc
      601 aagaatatca aagatacagt ctcagaagac caaagggcta ttgagacttt tcaacaaagg
      661 gtaatatcgg gaaacctcct cggattccat tgcccagcta tctgtcactt catcaaaagg
      721 acagtagaaa aggaaggtgg cacctacaaa tgccatcatt gcgataaagg aaaggctatc
      781 gttcaagatg cctctgccga cagtggtccc aaagatggac ccccacccac gaggagcatc
      841 gtggaaaaag aagacgttcc aaccacgtct tcaaagcaag tggattgatg tgatatctcc
      901 actgacgtaa gggatgacgc acaatcccac tatccttcgc aagacccttc ctctatataa
      961 ggaagttcat ttcatttgga gaggacacgc tgaaatcacc agtctctctc tacaaatcta
     1021 tctctctcga gctttcgcag atccgggggg caatgagata tgaaaaagcc tgaactcacc
     1081 gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga cctgatgcag
     1141 ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg tggatatgtc
     1201 ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta tcggcacttt
     1261 gcatcggccg cgctcccgat tccggaagtg cttgacattg gggagtttag cgagagcctg
     1321 acctattgca tctcccgccg ttcacagggt gtcacgttgc aagacctgcc tgaaaccgaa
     1381 ctgcccgctg ttctacaacc ggtcgcggag gctatggatg cgatcgctgc ggccgatctt
     1441 agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata cactacatgg
     1501 cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac tgtgatggac
     1561 gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac
     1621 tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac
     1681 aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac
     1741 gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc
     1801 tacttcgagc ggaggcatcc ggagcttgca ggatcgccac gactccgggc gtatatgctc
     1861 cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct
     1921 tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca
     1981 caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt actcgccgat
     2041 agtggaaacc gacgccccag cactcgtccg agggcaaaga aatagagtag atgccgaccg
     2101 ggatctgtcg atcgacaagc tcgagtttct ccataataat gtgtgagtag ttcccagata
     2161 agggaattag ggttcctata gggtttcgct catgtgttga gcatataaga aacccttagt
     2221 atgtatttgt atttgtaaaa tacttctatc aataaaattt ctaattccta aaaccaaaat
     2281 ccagtactaa aatccagatc ccccgaatta attcggcgtt aattcagtac attaaaaacg
     2341 tccgcaatgt gttattaagt tgtctaagcg tcaatttgtt tacaccacaa tatatcctgc
     2401 caccagccag ccaacagctc cccgaccggc agctcggcac aaaatcacca ctcgatacag
     2461 gcagcccatc agtccgggac ggcgtcagcg ggagagccgt tgtaaggcgg cagactttgc
     2521 tcatgttacc gatgctattc ggaagaacgg caactaagct gccgggtttg aaacacggat
     2581 gatctcgcgg agggtagcat gttgattgta acgatgacag agcgttgctg cctgtgatca
     2641 ccgcggtttc aaaatcggct ccgtcgatac tatgttatac gccaactttg aaaacaactt
     2701 tgaaaaagct gttttctggt atttaaggtt ttagaatgca aggaacagtg aattggagtt
     2761 cgtcttgtta taattagctt cttggggtat ctttaaatac tgtagaaaag aggaaggaaa
     2821 taataaatgg ctaaaatgag aatatcaccg gaattgaaaa aactgatcga aaaataccgc
     2881 tgcgtaaaag atacggaagg aatgtctcct gctaaggtat ataagctggt gggagaaaat
     2941 gaaaacctat atttaaaaat gacggacagc cggtataaag ggaccaccta tgatgtggaa
     3001 cgggaaaagg acatgatgct atggctggaa ggaaagctgc ctgttccaaa ggtcctgcac
     3061 tttgaacggc atgatggctg gagcaatctg ctcatgagtg aggccgatgg cgtcctttgc
     3121 tcggaagagt atgaagatga acaaagccct gaaaagatta tcgagctgta tgcggagtgc
     3181 atcaggctct ttcactccat cgacatatcg gattgtccct atacgaatag cttagacagc
     3241 cgcttagccg aattggatta cttactgaat aacgatctgg ccgatgtgga ttgcgaaaac
     3301 tgggaagaag acactccatt taaagatccg cgcgagctgt atgatttttt aaagacggaa
     3361 aagcccgaag aggaacttgt cttttcccac ggcgacctgg gagacagcaa catctttgtg
     3421 aaagatggca aagtaagtgg ctttattgat cttgggagaa gcggcagggc ggacaagtgg
     3481 tatgacattg ccttctgcgt ccggtcgatc agggaggata tcggggaaga acagtatgtc
     3541 gagctatttt ttgacttact ggggatcaag cctgattggg agaaaataaa atattatatt
     3601 ttactggatg aattgtttta gtacctagaa tgcatgacca aaatccctta acgtgagttt
     3661 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     3721 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3781 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     3841 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     3901 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     3961 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4021 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     4081 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     4141 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4201 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4261 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4321 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     4381 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     4441 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
     4501 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     4561 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
     4621 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     4681 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     4741 catcaccgaa acgcgcgagg cagggtgcct tgatgtgggc gccggcggtc gagtggcgac
     4801 ggcgcggctt gtccgcgccc tggtagattg cctggccgta ggccagccat ttttgagcgg
     4861 ccagcggccg cgataggccg acgcgaagcg gcggggcgta gggagcgcag cgaccgaagg
     4921 gtaggcgctt tttgcagctc ttcggctgtg cgctggccag acagttatgc acaggccagg
     4981 cgggttttaa gagttttaat aagttttaaa gagttttagg cggaaaaatc gccttttttc
     5041 tcttttatat cagtcactta catgtgtgac cggttcccaa tgtacggctt tgggttccca
     5101 atgtacgggt tccggttccc aatgtacggc tttgggttcc caatgtacgt gctatccaca
     5161 ggaaagagac cttttcgacc tttttcccct gctagggcaa tttgccctag catctgctcc
     5221 gtacattagg aaccggcgga tgcttcgccc tcgatcaggt tgcggtagcg catgactagg
     5281 atcgggccag cctgccccgc ctcctccttc aaatcgtact ccggcaggtc atttgacccg
     5341 atcagcttgc gcacggtgaa acagaacttc ttgaactctc cggcgctgcc actgcgttcg
     5401 tagatcgtct tgaacaacca tctggcttct gccttgcctg cggcgcggcg tgccaggcgg
     5461 tagagaaaac ggccgatgcc gggatcgatc aaaaagtaat cggggtgaac cgtcagcacg
     5521 tccgggttct tgccttctgt gatctcgcgg tacatccaat cagctagctc gatctcgatg
     5581 tactccggcc gcccggtttc gctctttacg atcttgtagc ggctaatcaa ggcttcaccc
     5641 tcggataccg tcaccaggcg gccgttcttg gccttcttcg tacgctgcat ggcaacgtgc
     5701 gtggtgttta accgaatgca ggtttctacc aggtcgtctt tctgctttcc gccatcggct
     5761 cgccggcaga acttgagtac gtccgcaacg tgtggacgga acacgcggcc gggcttgtct
     5821 cccttccctt cccggtatcg gttcatggat tcggttagat gggaaaccgc catcagtacc
     5881 aggtcgtaat cccacacact ggccatgccg gccggccctg cggaaacctc tacgtgcccg
     5941 tctggaagct cgtagcggat cacctcgcca gctcgtcggt cacgcttcga cagacggaaa
     6001 acggccacgt ccatgatgct gcgactatcg cgggtgccca cgtcatagag catcggaacg
     6061 aaaaaatctg gttgctcgtc gcccttgggc ggcttcctaa tcgacggcgc accggctgcc
     6121 ggcggttgcc gggattcttt gcggattcga tcagcggccg cttgccacga ttcaccgggg
     6181 cgtgcttctg cctcgatgcg ttgccgctgg gcggcctgcg cggccttcaa cttctccacc
     6241 aggtcatcac ccagcgccgc gccgatttgt accgggccgg atggtttgcg accgctcacg
     6301 ccgattcctc gggcttgggg gttccagtgc cattgcaggg ccggcaggca acccagccgc
     6361 ttacgcctgg ccaaccgccc gttcctccac acatggggca ttccacggcg tcggtgcctg
     6421 gttgttcttg attttccatg ccgcctcctt tagccgctaa aattcatcta ctcatttatt
     6481 catttgctca tttactctgg tagctgcgcg atgtattcag atagcagctc ggtaatggtc
     6541 ttgccttggc gtaccgcgta catcttcagc ttggtgtgat cctccgccgg caactgaaag
     6601 ttgacccgct tcatggctgg cgtgtctgcc aggctggcca acgttgcagc cttgctgctg
     6661 cgtgcgctcg gacggccggc acttagcgtg tttgtgcttt tgctcatttt ctctttacct
     6721 cattaactca aatgagtttt gatttaattt cagcggccag cgcctggacc tcgcgggcag
     6781 cgtcgccctc gggttctgat tcaagaacgg ttgtgccggc ggcggcagtg cctgggtagc
     6841 tcacgcgctg cgtgatacgg gactcaagaa tgggcagctc gtacccggcc agcgcctcgg
     6901 caacctcacc gccgatgcgc gtgcctttga tcgcccgcga cacgacaaag gccgcttgta
     6961 gccttccatc cgtgacctca atgcgctgct taaccagctc caccaggtcg gcggtggccc
     7021 atatgtcgta agggcttggc tgcaccggaa tcagcacgaa gtcggctgcc ttgatcgcgg
     7081 acacagccaa gtccgccgcc tggggcgctc cgtcgatcac tacgaagtcg cgccggccga
     7141 tggccttcac gtcgcggtca atcgtcgggc ggtcgatgcc gacaacggtt agcggttgat
     7201 cttcccgcac ggccgcccaa tcgcgggcac tgccctgggg atcggaatcg actaacagaa
     7261 catcggcccc ggcgagttgc agggcgcggg ctagatgggt tgcgatggtc gtcttgcctg
     7321 acccgccttt ctggttaagt acagcgataa ccttcatgcg ttccccttgc gtatttgttt
     7381 atttactcat cgcatcatat acgcagcgac cgcatgacgc aagctgtttt actcaaatac
     7441 acatcacctt tttagacggc ggcgctcggt ttcttcagcg gccaagctgg ccggccaggc
     7501 cgccagcttg gcatcagaca aaccggccag gatttcatgc agccgcacgg ttgagacgtg
     7561 cgcgggcggc tcgaacacgt acccggccgc gatcatctcc gcctcgatct cttcggtaat
     7621 gaaaaacggt tcgtcctggc cgtcctggtg cggtttcatg cttgttcctc ttggcgttca
     7681 ttctcggcgg ccgccagggc gtcggcctcg gtcaatgcgt cctcacggaa ggcaccgcgc
     7741 cgcctggcct cggtgggcgt cacttcctcg ctgcgctcaa gtgcgcggta cagggtcgag
     7801 cgatgcacgc caagcagtgc agccgcctct ttcacggtgc ggccttcctg gtcgatcagc
     7861 tcgcgggcgt gcgcgatctg tgccggggtg agggtagggc gggggccaaa cttcacgcct
     7921 cgggccttgg cggcctcgcg cccgctccgg gtgcggtcga tgattaggga acgctcgaac
     7981 tcggcaatgc cggcgaacac ggtcaacacc atgcggccgg ccggcgtggt ggtgtcggcc
     8041 cacggctctg ccaggctacg caggcccgcg ccggcctcct ggatgcgctc ggcaatgtcc
     8101 agtaggtcgc gggtgctgcg ggccaggcgg tctagcctgg tcactgtcac aacgtcgcca
     8161 gggcgtaggt ggtcaagcat cctggccagc tccgggcggt cgcgcctggt gccggtgatc
     8221 ttctcggaaa acagcttggt gcagccggcc gcgtgcagtt cggcccgttg gttggtcaag
     8281 tcctggtcgt cggtgctgac gcgggcatag cccagcaggc cagcggcggc gctcttgttc
     8341 atggcgtaat gtctccggtt ctagtcgcaa gtattctact ttatgcgact aaaacacgcg
     8401 acaagaaaac gccaggaaaa gggcagggcg gcagcctgtc gcgtaactta ggacttgtgc
     8461 gacatgtcgt tttcagaaga cggctgcact gaacgtcaga agccgactgc actatagcag
     8521 cggaggggtt ggatcaaagt actttgatcc cgaggggaac cctgtggttg gcatgcacat
     8581 acaaatggac gaacggataa accttttcac gcccttttaa atatccgatt attctaataa
     8641 acgctctttt ctcttaggtt tacccgccaa tatatcctgt caaacactga tagtttaaac
     8701 tgaaggcggg aaacgacaat ctgatccaag ctcaagctgc tctagcattc gccattcagg
     8761 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     8821 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     8881 cgttgtaaaa cgacggccag tgccaagctt gattacgaat tcgagctcgg tacccgacga
     8941 gtcagtaata aacggcgtca aagtggttgc agccggcaca cacgagtcgt gtttatcaac
     9001 tcaaagcaca aatacttttc ctcaacctaa aaataaggca attagccaaa aacaactttg
     9061 cgtgtaaaca acgctcaata cacgtgtcat tttattatta gctattgctt caccgcctta
     9121 gctttctcgt gacctagtcg tcctcgtctt ttcttcttct tcttctataa aacaataccc
     9181 aaagagctct tcttcttcac aattcagatt tcaatttctc aaaatcttaa aaactttctc
     9241 tcaattctct ctaccgtgat caaggtaaat ttctgtgttc cttattctct caaaatcttc
     9301 gattttgttt tcgttcgatc ccaatttcgt atatgttctt tggtttagat tctgttaatc
     9361 ttagatcgaa gacgattttc tgggtttgat cgttagatat catcttaatt ctcgattagg
     9421 gtttcataga tatcatccga tttgttcaaa taatttgagt tttgtcgaat aattactctt
     9481 cgatttgtga tttctatcta gatctggtgt tagtttctag tttgtgcgat cgaatttgta
     9541 gattaatctg agtttttctg attaacagct cgagtgcggg ccccccctcg aggcgcgcca
     9601 agctatcaaa caagtttgta caaaaaagca ggctaccata tggccagcag cccccccaag
     9661 aagaagcgca aggtcgacaa gaagtactcc atcggcctgg acatcggcac gaacagcgtg
     9721 ggctgggccg tgatcaccga cgagtacaag gtgccgtcca agaagttcaa ggtgctgggc
     9781 aacaccgacc gccacagcat caagaagaac ctgatcggcg ccctgctgtt cgactcgggc
     9841 gagaccgccg aggcgacgcg cctgaagcgc accgcgcgcc gccgctacac gcgccgcaag
     9901 aaccgcatct gctacctgca ggagatcttc agcaacgaga tggccaaggt ggacgactcg
     9961 ttcttccacc gcctggagga gtccttcctg gtggaggagg acaagaagca cgagcgccac
    10021 cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc gacgatctac
    10081 cacctgcgca agaagctggt ggacagcacc gacaaggcgg acctgcgcct gatctacctg
    10141 gccctggcgc acatgatcaa gttccgcggc cacttcctga tcgagggcga cctgaacccc
    10201 gacaactcgg acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc
    10261 gaggagaacc cgatcaacgc ctcgggcgtg gacgccaagg cgatcctgtc cgcgcgcctg
    10321 tccaagagcc gccgcttgga gaacctgatc gcccagctgc ccggcgagaa gaagaacggc
    10381 ctgttcggca acctgatcgc gctgagcctg ggcctgacgc cgaacttcaa gtcgaacttc
    10441 gacctggccg aggacgcgaa gctgcagctg agcaaggaca cctacgacga cgacctggac
    10501 aacctgctgg cccagatcgg cgaccagtac gcggacctgt tcctggccgc gaagaacctg
    10561 tcggacgcca tcctgctgtc cgacatcctg cgcgtgaaca ccgagatcac gaaggccccc
    10621 ctgtcggcgt ccatgatcaa gcgctacgac gagcaccacc aggacctgac cctgctgaag
    10681 gcgctggtgc gccagcagct gccggagaag tacaaggaga tcttcttcga ccagtccaag
    10741 aacggctacg ccggctacat cgacggcggc gccagccagg aggagttcta caagttcatc
    10801 aagcccatcc tggagaagat ggacggcacg gaggagctgc tggtgaagct gaaccgcgag
    10861 gacctgctgc gcaagcagcg caccttcgac aacggcagca tcccccacca gatccacctg
    10921 ggcgagctgc acgccatcct gcgccgccag gaggacttct acccgttcct gaaggacaac
    10981 cgcgagaaga tcgagaagat cctgacgttc cgcatcccct actacgtggg cccgctggct
    11041 cgcggcaact cccgcttcgc gtggatgacc cgcaagagcg aggagaccat cacgccctgg
    11101 aacttcgagg aggtggtgga caagggcgcc agcgcgcagt cgttcatcga gcgcatgacc
    11161 aacttcgaca agaacctgcc caacgagaag gtgctgccga agcactccct gctgtacgag
    11221 tacttcaccg tgtacaacga gctgacgaag gtgaagtacg tgaccgaggg catgcgcaag
    11281 cccgccttcc tgagcggcga gcagaagaag gcgatcgtgg acctgctgtt caagaccaac
    11341 cgcaaggtga cggtgaagca gctgaaggag gactacttca agaagatcga gtgcttcgac
    11401 agcgtggaga tctcgggcgt ggaggaccgc ttcaacgcca gcctgggcac ctaccacgac
    11461 ctgctgaaga tcatcaagga caaggacttc ctggacaacg aggagaacga ggacatcctg
    11521 gaggacatcg tgctgaccct gacgctgttc gaggaccgcg agatgatcga ggagcgcctg
    11581 aagacgtacg cccacctgtt cgacgacaag gtgatgaagc agctgaagcg ccgccgctac
    11641 accggctggg gccgcctgag ccgcaagctg atcaacggca tccgcgacaa gcagtcgggc
    11701 aagaccatcc tggacttcct gaagtccgac ggcttcgcga accgcaactt catgcagctg
    11761 atccacgacg actcgctgac cttcaaggag gacatccaga aggcccaggt gtcgggccag
    11821 ggcgactccc tgcacgagca catcgccaac ctggcgggct cccccgcgat caagaagggc
    11881 atcctgcaga ccgtgaaggt ggtggacgag ctggtgaagg tgatgggccg ccacaagccg
    11941 gagaacatcg tgatcgagat ggcccgcgag aaccagacca cgcagaaggg ccagaagaac
    12001 agccgcgagc gcatgaagcg catcgaggag ggcatcaagg agctgggctc gcagatcctg
    12061 aaggagcacc ccgtggagaa cacccagctg cagaacgaga agctgtacct gtactacctg
    12121 cagaacggcc gcgacatgta cgtggaccag gagctggaca tcaaccgcct gtccgactac
    12181 gacgtggacc acatcgtgcc ccagagcttc ctgaaggacg actcgatcga caacaaggtg
    12241 ctgacccgca gcgacaagaa ccgcggcaag agcgacaacg tgccgtcgga ggaggtggtg
    12301 aagaagatga agaactactg gcgccagctg ctgaacgcca agctgatcac gcagcgcaag
    12361 ttcgacaacc tgaccaaggc cgagcgcggc ggcctgtcgg agctggacaa ggcgggcttc
    12421 atcaagcgcc agctggtgga gacccgccag atcacgaagc acgtggcgca gatcctggac
    12481 tcccgcatga acacgaagta cgacgagaac gacaagctga tccgcgaggt gaaggtgatc
    12541 accctgaagt ccaagctggt cagcgacttc cgcaaggact tccagttcta caaggtgcgc
    12601 gagatcaaca actaccacca cgcccacgac gcgtacctga acgccgtggt gggcaccgcg
    12661 ctgatcaaga agtaccccaa gctggagagc gagttcgtgt acggcgacta caaggtgtac
    12721 gacgtgcgca agatgatcgc caagtcggag caggagatcg gcaaggccac cgcgaagtac
    12781 ttcttctact ccaacatcat gaacttcttc aagaccgaga tcacgctggc caacggcgag
    12841 atccgcaagc gccccctgat cgagaccaac ggcgagacgg gcgagatcgt gtgggacaag
    12901 ggccgcgact tcgcgaccgt gcgcaaggtg ctgagcatgc cccaggtgaa catcgtgaag
    12961 aagaccgagg tgcagacggg cggcttctcc aaggagagca tcctgccgaa gcgcaactcg
    13021 gacaagctga tcgcccgcaa gaaggactgg gaccccaaga agtacggcgg cttcgactcc
    13081 ccgaccgtgg cctacagcgt gctggtggtg gcgaaggtgg agaagggcaa gtccaagaag
    13141 ctgaagagcg tgaaggagct gctgggcatc accatcatgg agcgcagctc gttcgagaag
    13201 aaccccatcg acttcctgga ggcgaagggc tacaaggagg tgaagaagga cctgatcatc
    13261 aagctgccga agtactcgct gttcgagctg gagaacggcc gcaagcgcat gctggcctcc
    13321 gcgggcgagc tgcagaaggg caacgagctg gccctgccca gcaagtacgt gaacttcctg
    13381 tacctggcgt cccactacga gaagctgaag ggctcgccgg aggacaacga gcagaagcag
    13441 ctgttcgtgg agcagcacaa gcactacctg gacgagatca tcgagcagat ctcggagttc
    13501 tccaagcgcg tgatcctggc cgacgcgaac ctggacaagg tgctgagcgc ctacaacaag
    13561 caccgcgaca agcccatccg cgagcaggcg gagaacatca tccacctgtt caccctgacg
    13621 aacctgggcg ccccggccgc gttcaagtac ttcgacacca cgatcgaccg caagcgctac
    13681 accagcacga aggaggtgct ggacgcgacc ctgatccacc agagcatcac cggcctgtac
    13741 gagacgcgca tcgacctgtc gcagctgggc ggcgactaat gtacccagct ttcttgtaca
    13801 aagtggttcg ataattctta attaactagt tctagagcgg ccgccaccgc ggtggagctc
    13861 gaatttcccc gatcgttcaa acatttggca ataaagtttc ttaagattga atcctgttgc
    13921 cggtcttgcg atgattatca tataatttct gttgaattac gttaagcatg taataattaa
    13981 catgtaatgc atgacgttat ttatgagatg ggtttttatg attagagtcc cgcaattata
    14041 catttaatac gcgatagaaa acaaaatata gcgcgcaaac taggataaat tatcgcgcgc
    14101 ggtgtcatct atgttactag atcgggaa
//

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