pHAtC

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: pHAtC 
出品公司: 爱迪基因
目录编号: 78098
存储实验室: Jin-Soo Kim

载体骨架基本信息
载体名称: pH2GW7
载体出品公司: --
载体抗性: 大观霉素
空载体大小: --
完整质粒大小: --
载体修饰: 在U6启动子驱动的sgRNA盒中,人类密码子优化了包含核定位信号(NLS)的SpCas9基因,并在SpCas9的C端缺失一个HA标签
载体类型: 植物表达,CRISPR
筛选标记: 潮霉素
高拷贝/低拷贝: --
生长条件: 37 ℃

插入基因信息
插入基因名称: --
物种来源: --
插入大小: --
基因突变: --
GenBank编号 --
启动子 U6
标签/融合蛋白: --
克隆方法: 限制性内切酶
5' 克隆位点: --
3' 克隆位点: --
5' 测序引物 --
3' 测序引物 --

文献引用方法
材料和方法部分: pHAtC was a gift from Jin-Soo Kim (Addgene plasmid # 78098 ; http://n2t.net/addgene:78098 ; RRID:Addgene_78098)
参考文献部分: A simple, flexible and high-throughput cloning system for plant genome editing via CRISPR-Cas system. Kim H, Kim ST, Ryu J, Choi MK, Kweon J, Kang BC, Ahn HM, Bae S, Kim JS, Kim SG. J Integr Plant Biol. 2016 Mar 4. doi: 10.1111/jipb.12474. 10.1111/jipb.12474 PubMed 26946469

载体图谱质粒图谱和多克隆位点信息

pHAtC质粒图谱

载体简介

载体序列

LOCUS       pHAtC               14451 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  Plant expression of Cas9 and gRNA empty backbone; Hygro Resistance.
ACCESSION   .
VERSION     .
KEYWORDS    pHAtC
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14451)
  AUTHORS   Kim H, Kim ST, Ryu J, Choi MK, Kweon J, Kang BC, Ahn HM, Bae S, Kim 
            JS, Kim SG
  TITLE     A simple, flexible and high-throughput cloning system for plant 
            genome editing via CRISPR-Cas system.
  JOURNAL   J Integr Plant Biol. 2016 Mar 4. doi: 10.1111/jipb.12474.
  PUBMED    26946469
REFERENCE   2  (bases 1 to 14451)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..14451
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_RNA        complement(13..88)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        1005..1349
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     1293..1313
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     protein_bind    1449..1473
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1476..5579
                     /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             5595..5615
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5616..5642
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(5647..5671)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    5991..6015
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             7346..7975
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             8404..9477
                     /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      9543..9737
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(9973..9995)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    10081..10221
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     10136..10155
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(10236..10253)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10407..10995)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10487..10506)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(11241..12032)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_feature    12553..12577
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      12670..12922
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(13120..14145)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(14181..14360)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
ORIGIN
        1 gagctcaaaa aagcaccgac tcggtgccac tttttcaagt tgataacgga ctagccttat
       61 tttaacttgc tatttctagc tctaaaacct ctgcaggtga ctctcgagac acctgcctcc
      121 aatcactact tcgactctag ctgtatataa actcagcttc gttttcttat ctaagcgatg
      181 tgggactttt gaagattgtt ttcaacttaa atgggcctat ataagaaata ctattgttct
      241 ttcccatata aatgggcctg cttctcttct ttcagattcc caggggcctt ttgaagatta
      301 tcttcatatc ttaagaatga agatgtttta ttcaatcaaa ttcttgaagg ttcgatgcct
      361 aatcattcga attcgagctc tcccatatgg tcgactagag ccaagctgat ctcctttgcc
      421 ccggagatca ccatggacga ctttctctat ctctacgatc taggaagaaa gttcgacgga
      481 gaaggtgacg ataccatgtt caccaccgat aatgagaaga ttagcctctt caatttcaga
      541 aagaatgctg acccacagat ggttagagag gcctacgcgg caggtctcat caagacgatc
      601 tacccgagta ataatctcca ggagatcaaa taccttccca agaaggttaa agatgcagtc
      661 aaaagattca ggactaactg catcaagaac acagagaaag atatatttct caagatcaga
      721 agtactattc cagtatggac gattcaaggc ttgcttcata aaccaaggca agtaatagag
      781 attggagtct ctaagaaagt agttcctact gaatcaaagg ccatggagtc aaaaattcag
      841 atcgaggatc taacagaact cgccgtgaag actggcgaac agttcataca gagtctttta
      901 cgactcaatg acaagaagaa aatcttcgtc aacatggtgg agcacgacac tctcgtctac
      961 tccaagaata tcaaagatac agtctcagaa gaccaaaggg ctattgagac ttttcaacaa
     1021 agggtaatat cgggaaacct cctcggattc cattgcccag ctatctgtca cttcatcaaa
     1081 aggacagtag aaaaggaagg tggcacctac aaatgccatc attgcgataa aggaaaggct
     1141 atcgttcaag atgcccctgc cgacagtggt cccaaagatg gacccccacc cacgaggagc
     1201 atcgtggaaa aagaagacgt tccaaccacg tcttcaaagc aagtggattg atgtgatatc
     1261 tccactgacg taagggatga cgcacaatcc cactatcctt cgcaagaccc ttcctctata
     1321 taaggaagtt catttcattt ggagaggact ccggtatttt tacaacaatt accacaacaa
     1381 aacaaacaac aaacaacatt acaatttact attctagtcg acctgcaggc ggccgcacta
     1441 gtgatatcac aagtttgtac aaaaaagcag gcttcatgga caagaagtac agcatcggcc
     1501 tggacatcgg taccaacagc gtgggctggg ccgtgatcac cgacgagtac aaggtgccca
     1561 gcaagaagtt caaggtgctg ggcaacaccg accgccacag catcaagaag aacctgatcg
     1621 gcgccctgct gttcgacagc ggcgagaccg ccgaggccac ccgcctgaag cgcaccgccc
     1681 gccgccgcta cacccgccgc aagaaccgca tctgctacct gcaggagatc ttcagcaacg
     1741 agatggccaa ggtggacgac agcttcttcc accgcctgga ggagagcttc ctggtggagg
     1801 aggacaagaa gcacgagcgc caccccatct tcggcaacat cgtggacgag gtggcctacc
     1861 acgagaagta ccccaccatc taccatctgc gcaagaagct ggtggacagc accgacaagg
     1921 ccgacctgcg cctgatctac ctggccctgg cccacatgat caagttccgc ggccacttcc
     1981 tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg
     2041 tgcagaccta caaccagctg ttcgaggaga accccatcaa cgccagcggc gtggacgcca
     2101 aggccatcct gagcgcccgc ctgagcaaga gccgccgcct ggagaacctg atcgcccagc
     2161 tgcccggcga gaagaagaac ggcctgttcg gcaacctgat cgccctgagc ctgggcctga
     2221 cccccaactt caagagcaac ttcgacctgg ccgaggacgc caagctgcag ctgagcaagg
     2281 acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc
     2341 tgttcctggc cgccaagaac ctgagcgacg ccatcctgct gagcgacatc ctgcgcgtga
     2401 acaccgagat caccaaggcc cccctgagcg ccagcatgat caagcgctac gacgagcacc
     2461 accaggacct gaccctgctg aaggccctgg tgcgccagca gctgcccgag aagtacaagg
     2521 agatcttctt cgaccagagc aagaacggct acgccggcta catcgacggc ggcgccagcc
     2581 aggaggagtt ctacaagttc atcaagccca tcctggagaa gatggacggc accgaggagc
     2641 tgctggtgaa gctgaaccgc gaggacctgc tgcgcaagca gcgcaccttc gacaacggca
     2701 gcatccccca ccagatccac ctgggcgagc tgcacgccat cctgcgccgc caggaggact
     2761 tctacccctt cctgaaggac aaccgcgaga agatcgagaa gatcctgacc ttccgcatcc
     2821 cctactacgt gggccccctg gcccgcggca acagccgctt cgcctggatg acccgcaaga
     2881 gcgaggagac catcaccccc tggaacttcg aggaggtggt ggacaagggc gccagcgccc
     2941 agagcttcat cgagcgcatg accaacttcg acaagaacct gcccaacgag aaggtgctgc
     3001 ccaagcacag cctgctgtac gagtacttca ccgtgtacaa cgagctgacc aaggtgaagt
     3061 acgtgaccga gggcatgcgc aagcccgcct tcctgagcgg cgagcagaag aaggccatcg
     3121 tggacctgct gttcaagacc aaccgcaagg tgaccgtgaa gcagctgaag gaggactact
     3181 tcaagaagat cgagtgcttc gacagcgtgg agatcagcgg cgtggaggac cgcttcaacg
     3241 ccagcctggg cacctaccac gacctgctga agatcatcaa ggacaaggac ttcctggaca
     3301 acgaggagaa cgaggacatc ctggaggaca tcgtgctgac cctgaccctg ttcgaggacc
     3361 gcgagatgat cgaggagcgc ctgaagacct acgcccacct gttcgacgac aaggtgatga
     3421 agcagctgaa gcgccgccgc tacaccggct ggggccgcct gagccgcaag cttatcaacg
     3481 gcatccgcga caagcagagc ggcaagacca tcctggactt cctgaagagc gacggcttcg
     3541 ccaaccgcaa cttcatgcag ctgatccacg acgacagcct gaccttcaag gaggacatcc
     3601 agaaggccca ggtgagcggc cagggcgaca gcctgcacga gcacatcgcc aacctggccg
     3661 gcagccccgc catcaagaag ggcatcctgc agaccgtgaa ggtggtggac gagctggtga
     3721 aggtgatggg ccgccacaag cccgagaaca tcgtgatcga gatggcccgc gagaaccaga
     3781 ccacccagaa gggccagaag aacagccgcg agcgcatgaa gcgcatcgag gagggcatca
     3841 aggagctggg cagccagatc ctgaaggagc accccgtgga gaacacccag ctgcagaacg
     3901 agaagctgta cctgtactac ctgcagaacg gccgcgacat gtacgtggac caggagctgg
     3961 acatcaaccg cctgagcgac tacgacgtgg accacatcgt gccccagagc ttcctgaagg
     4021 acgacagcat cgacaacaag gtgctgaccc gcagcgacaa gaaccgcggc aagagcgaca
     4081 acgtgcccag cgaggaggtg gtgaagaaga tgaagaacta ctggcgccag ctgctgaacg
     4141 ccaagctgat cacccagcgc aagttcgaca acctgaccaa ggccgagcgc ggcggcctga
     4201 gcgagctgga caaggccggc ttcatcaagc gccagctggt ggagacccgc cagatcacca
     4261 agcacgtggc ccagatcctg gacagccgca tgaacaccaa gtacgacgag aacgacaagc
     4321 tgatccgcga ggtgaaggtg atcaccctga agagcaagct ggtgagcgac ttccgcaagg
     4381 acttccagtt ctacaaggtg cgcgagatca acaactacca ccacgcccac gacgcctacc
     4441 tgaacgccgt ggtgggcacc gccctgatca agaagtaccc caagctggag agcgagttcg
     4501 tgtacggcga ctacaaggtg tacgacgtgc gcaagatgat cgccaagagc gagcaggaga
     4561 tcggcaaggc caccgccaag tacttcttct acagcaacat catgaacttc ttcaagaccg
     4621 agatcaccct ggccaacggc gagatccgca agcgccccct gatcgagacc aacggcgaga
     4681 ccggcgagat cgtgtgggac aagggccgcg acttcgccac cgtgcgcaag gtgctgagca
     4741 tgccccaggt gaacatcgtg aagaagaccg aggtgcagac cggcggcttc agcaaggaga
     4801 gcatcctgcc caagcgcaac agcgacaagc tgatcgcccg caagaaggac tgggacccca
     4861 agaagtacgg cggcttcgac agccccaccg tggcctacag cgtgctggtg gtggccaagg
     4921 tggagaaggg caagagcaag aagctgaaga gcgtgaagga gctgctgggc atcaccatca
     4981 tggagcgcag cagcttcgag aagaacccca tcgacttcct ggaggccaag ggctacaagg
     5041 aggtgaagaa ggacctgatc atcaagctgc ccaagtacag cctgttcgag ctggagaacg
     5101 gccgcaagcg catgctggcc agcgccggcg agctgcagaa gggcaacgag ctggccctgc
     5161 ccagcaagta cgtgaacttc ctgtacctgg ccagccacta cgagaagctg aagggcagcc
     5221 ccgaggacaa cgagcagaag cagctgttcg tggagcagca caagcactac ctggacgaga
     5281 tcatcgagca gatcagcgag ttcagcaagc gcgtgatcct ggccgacgcc aacctggaca
     5341 aggtgctgag cgcctacaac aagcaccgcg acaagcccat ccgcgagcag gccgagaaca
     5401 tcatccacct gttcaccctg accaacctgg gcgcccccgc cgccttcaag tacttcgaca
     5461 ccaccatcga ccgcaagcgc tacaccagca ccaaggaggt gctggacgcc accctgatcc
     5521 accagagcat caccggtctg tacgagaccc gcatcgacct gagccagctg ggcggcgacg
     5581 gcggctccgg acctccaaag aaaaagagaa aagtataccc ctacgacgtg cccgactacg
     5641 cctaacaccc agctttcttg tacaaagtgg tgatatcccg cggccatgct agagtccgca
     5701 aaaatcacca gtctctctct acaaatctat ctctctctat ttttctccag aataatgtgt
     5761 gagtagttcc cagataaggg aattagggtt cttatagggt ttcgctcatg tgttgagcat
     5821 ataagaaacc cttagtatgt atttgtattt gtaaaatact tctatcaata aaatttctaa
     5881 ttcctaaaac caaaatccag tgacctgcag gcatgcgacg tcgggcccaa gcttagcttg
     5941 agcttggatc agattgtcgt ttcccgcctt cagtttaaac tatcagtgtt tgacaggata
     6001 tattggcggg taaacctaag agaaaagagc gtttattaga ataatcggat atttaaaagg
     6061 gcgtgaaaag gtttatccgt tcgtccattt gtatgtgcat gccaaccaca gggttcccct
     6121 cgggatcaaa gtactttaaa gtactttaaa gtactttaaa gtactttgat ccaacccctc
     6181 cgctgctata gtgcagtcgg cttctgacgt tcagtgcagc cgtcttctga aaacgacatg
     6241 tcgcacaagt cctaagttac gcgacaggct gccgccctgc ccttttcctg gcgttttctt
     6301 gtcgcgtgtt ttagtcgcat aaagtagaat acttgcgact agaaccggag acattacgcc
     6361 atgaacaaga gcgccgccgc tggcctgctg ggctatgccc gcgtcagcac cgacgaccag
     6421 gacttgacca accaacgggc cgaactgcac gcggccggct gcaccaagct gttttccgag
     6481 aagatcaccg gcaccaggcg cgaccgcccg gagctggcca ggatgcttga ccacctacgc
     6541 cctggcgacg ttgtgacagt gaccaggcta gaccgcctgg cccgcagcac ccgcgaccta
     6601 ctggacattg ccgagcgcat ccaggaggcc ggcgcgggcc tgcgtagcct ggcagagccg
     6661 tgggccgaca ccaccacgcc ggccggccgc atggtgttga ccgtgttcgc cggcattgcc
     6721 gagttcgagc gttccctaat catcgaccgc acccggagcg ggcgcgaggc cgccaaggcc
     6781 cgaggcgtga agtttggccc ccgccctacc ctcaccccgg cacagatcgc gcacgcccgc
     6841 gagctgatcg accaggaagg ccgcaccgtg aaagaggcgg ctgcactgct tggcgtgcat
     6901 cgctcgaccc tgtaccgcgc acttgagcgc agcgaggaag tgacgcccac cgaggccagg
     6961 cggcgcggtg ccttccgtga ggacgcattg accgaggccg acgccctggc ggccgccgag
     7021 aatgaacgcc aagaggaaca agcatgaaac cgcaccagga cggccaggac gaaccgtttt
     7081 tcattaccga agagatcgag gcggagatga tcgcggccgg gtacgtgttc gagccgcccg
     7141 cgcacgtctc aaccgtgcgg ctgcatgaaa tcctggccgg tttgtctgat gccaagctgg
     7201 cggcctggcc ggccagcttg gccgctgaag aaaccgagcg ccgccgtcta aaaaggtgat
     7261 gtgtatttga gtaaaacagc ttgcgtcatg cggtcgctgc gtatatgatg cgatgagtaa
     7321 ataaacaaat acgcaagggg aacgcatgaa ggttatcgct gtacttaacc agaaaggcgg
     7381 gtcaggcaag acgaccatcg caacccatct agcccgcgcc ctgcaactcg ccggggccga
     7441 tgttctgtta gtcgattccg atccccaggg cagtgcccgc gattgggcgg ccgtgcggga
     7501 agatcaaccg ctaaccgttg tcggcatcga ccgcccgacg attgaccgcg acgtgaaggc
     7561 catcggccgg cgcgacttcg tagtgatcga cggagcgccc caggcggcgg acttggctgt
     7621 gtccgcgatc aaggcagccg acttcgtgct gattccggtg cagccaagcc cttacgacat
     7681 atgggccacc gccgacctgg tggagctggt taagcagcgc attgaggtca cggatggaag
     7741 gctacaagcg gcctttgtcg tgtcgcgggc gatcaaaggc acgcgcatcg gcggtgaggt
     7801 tgccgaggcg ctggccgggt acgagctgcc cattcttgag tcccgtatca cgcagcgcgt
     7861 gagctaccca ggcactgccg ccgccggcac aaccgttctt gaatcagaac ccgagggcga
     7921 cgctgcccgc gaggtccagg cgctggccgc tgaaattaaa tcaaaactca tttgagttaa
     7981 tgaggtaaag agaaaatgag caaaagcaca aacacgctaa gtgccggccg tccgagcgca
     8041 cgcagcagca aggctgcaac gttggccagc ctggcagaca cgccagccat gaagcgggtc
     8101 aactttcagt tgccggcgga ggatcacacc aagctgaaga tgtacgcggt acgccaaggc
     8161 aagaccatta ccgagctgct atctgaatac atcgcgcagc taccagagta aatgagcaaa
     8221 tgaataaatg agtagatgaa ttttagcggc taaaggaggc ggcatggaaa atcaagaaca
     8281 accaggcacc gacgccgtgg aatgccccat gtgtggagga acgggcggtt ggccaggcgt
     8341 aagcggctgg gttgtctgcc ggccctgcaa tggcactgga acccccaagc ccgaggaatc
     8401 ggcgtgagcg gtcgcaaacc atccggcccg gtacaaatcg gcgcggcgct gggtgatgac
     8461 ctggtggaga agttgaaggc cgcgcaggcc gcccagcggc aacgcatcga ggcagaagca
     8521 cgccccggtg aatcgtggca agcggccgct gatcgaatcc gcaaagaatc ccggcaaccg
     8581 ccggcagccg gtgcgccgtc gattaggaag ccgcccaagg gcgacgagca accagatttt
     8641 ttcgttccga tgctctatga cgtgggcacc cgcgatagtc gcagcatcat ggacgtggcc
     8701 gttttccgtc tgtcgaagcg tgaccgacga gctggcgagg tgatccgcta cgagcttcca
     8761 gacgggcacg tagaggtttc cgcagggccg gccggcatgg ccagtgtgtg ggattacgac
     8821 ctggtactga tggcggtttc ccatctaacc gaatccatga accgataccg ggaagggaag
     8881 ggagacaagc ccggccgcgt gttccgtcca cacgttgcgg acgtactcaa gttctgccgg
     8941 cgagccgatg gcggaaagca gaaagacgac ctggtagaaa cctgcattcg gttaaacacc
     9001 acgcacgttg ccatgcagcg tacgaagaag gccaagaacg gccgcctggt gacggtatcc
     9061 gagggtgaag ccttgattag ccgctacaag atcgtaaaga gcgaaaccgg gcggccggag
     9121 tacatcgaga tcgagctagc tgattggatg taccgcgaga tcacagaagg caagaacccg
     9181 gacgtgctga cggttcaccc cgattacttt ttgatcgatc ccggcatcgg ccgttttctc
     9241 taccgcctgg cacgccgcgc cgcaggcaag gcagaagcca gatggttgtt caagacgatc
     9301 tacgaacgca gtggcagcgc cggagagttc aagaagttct gtttcaccgt gcgcaagctg
     9361 atcgggtcaa atgacctgcc ggagtacgat ttgaaggagg aggcggggca ggctggcccg
     9421 atcctagtca tgcgctaccg caacctgatc gagggcgaag catccgccgg ttcctaatgt
     9481 acggagcaga tgctagggca aattgcccta gcaggggaaa aaggtcgaaa aggtctcttt
     9541 cctgtggata gcacgtacat tgggaaccca aagccgtaca ttgggaaccg gaacccgtac
     9601 attgggaacc caaagccgta cattgggaac cggtcacaca tgtaagtgac tgatataaaa
     9661 gagaaaaaag gcgatttttc cgcctaaaac tctttaaaac ttattaaaac tcttaaaacc
     9721 cgcctggcct gtgcataact gtctggccag cgcacagccg aagagctgca aaaagcgcct
     9781 acccttcggt cgctgcgctc cctacgcccc gccgcttcgc gtcggcctat cgcggccgct
     9841 ggccgctcaa aaatggctgg cctacggcca ggcaatctac cagggcgcgg acaagccgcg
     9901 ccgtcgccac tcgaccgccg gcgcccacat caaggcaccc tgcctcgcgc gtttcggtga
     9961 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
    10021 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
    10081 cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca
    10141 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta
    10201 aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
    10261 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
    10321 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
    10381 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
    10441 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
    10501 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
    10561 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
    10621 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
    10681 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
    10741 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
    10801 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
    10861 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
    10921 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
    10981 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
    11041 gaaaactcac gttaagggat tttggtcatg catgatatat ctcccaattt gtgtagggct
    11101 tattatgcac gcttaaaaat aataaaagca gacttgacct gatagtttgg ctgtgagcaa
    11161 ttatgtgctt agtgcatcta acgcttgagt taagccgcgc cgcgaagcgg cgtcggcttg
    11221 aacgaatttc tagctagaca ttatttgccg actaccttgg tgatctcgcc tttcacgtag
    11281 tggacaaatt cttccaactg atctgcgcgc gaggccaagc gatcttcttc ttgtccaaga
    11341 taagcctgtc tagcttcaag tatgacgggc tgatactggg ccggcaggcg ctccattgcc
    11401 cagtcggcag cgacatcctt cggcgcgatt ttgccggtta ctgcgctgta ccaaatgcgg
    11461 gacaacgtaa gcactacatt tcgctcatcg ccagcccagt cgggcggcga gttccatagc
    11521 gttaaggttt catttagcgc ctcaaataga tcctgttcag gaaccggatc aaagagttcc
    11581 tccgccgctg gacctaccaa ggcaacgcta tgttctcttg cttttgtcag caagatagcc
    11641 agatcaatgt cgatcgtggc tggctcgaag atacctgcaa gaatgtcatt gcgctgccat
    11701 tctccaaatt gcagttcgcg cttagctgga taacgccacg gaatgatgtc gtcgtgcaca
    11761 acaatggtga cttctacagc gcggagaatc tcgctctctc caggggaagc cgaagtttcc
    11821 aaaaggtcgt tgatcaaagc tcgccgcgtt gtttcatcaa gccttacggt caccgtaacc
    11881 agcaaatcaa tatcactgtg tggcttcagg ccgccatcca ctgcggagcc gtacaaatgt
    11941 acggccagca acgtcggttc gagatggcgc tcgatgacgc caactacctc tgatagttga
    12001 gtcgatactt cggcgatcac cgcttccccc atgatgttta actttgtttt agggcgactg
    12061 ccctgctgcg taacatcgtt gctgctccat aacatcaaac atcgacccac ggcgtaacgc
    12121 gcttgctgct tggatgcccg aggcatagac tgtaccccaa aaaaacagtc ataacaagcc
    12181 atgaaaaccg ccactgcgcc gttaccaccg ctgcgttcgg tcaaggttct ggaccagttg
    12241 cgtgagcgca tacgctactt gcattacagc ttacgaaccg aacaggctta tgtccactgg
    12301 gttcgtgccc gaattgatca caggcagcaa cgctctgtca tcgttacaat caacatgcta
    12361 ccctccgcga gatcatccgt gtttcaaacc cggcagctta gttgccgttc ttccgaatag
    12421 catcggtaac atgagcaaag tctgccgcct tacaacggct ctcccgctga cgccgtcccg
    12481 gactgatggg ctgcctgtat cgagtggtga ttttgtgccg agctgccggt cggggagctg
    12541 ttggctggct ggtggcagga tatattgtgg tgtaaacaaa ttgacgctta gacaacttaa
    12601 taacacattg cggacgtttt taatgtactg aattaacgcc gaattgaatt atcagcttgc
    12661 atgccggtcg atctagtaac atagatgaca ccgcgcgcga taatttatcc tagtttgcgc
    12721 gctatatttt gttttctatc gcgtattaaa tgtataattg cgggactcta atcataaaaa
    12781 cccatctcat aaataacgtc atgcattaca tgttaattat tacatgctta acgtaattca
    12841 acagaaatta tatgataatc atcgcaagac cggcaacagg attcaatctt aagaaacttt
    12901 attgccaaat gtttgaacga tctgcttgac tctagggaat taattcctga atcactgcga
    12961 ccggccctcc cgcgacccag ccgagcgagc ttagcgaact gtggacgaga actgtgccac
    13021 caagcgtaag gccgttctct cgcatttgcc ttgctaggct cgcgcgagtt gctggctgag
    13081 gcgttctcga aatcagctct tgttcggtcg gcatctactc tattcctttg ccctcggacg
    13141 agtgctgggg cgtcggtttc cactatcggc gagtacttct acacagccat cggtccagac
    13201 ggccgcgctt ctgcgggcga tttgtgtacg cccgacagtc ccggctccgg atcggacgat
    13261 tgcgtcgcat cgaccctgcg cccaagctgc atcatcgaaa ttgccgtcaa ccaagctctg
    13321 atagagttgg tcaagaccaa tgcggagcat atacgcccgg agccttggcg atcctgcaag
    13381 ctccggatgc ctccgctcga agtagcgcgt ctgctgctcc atacaagcca accacggcct
    13441 ccagaagaag atgttggcga cctcgtattg ggaatccccg aacatcgcct cgctccagtc
    13501 aatgaccgct gttatgcggc cattgtccgt caggacattg ttggagccga aatccgcgtg
    13561 cacgaggtgc cggacttcgg ggcagtcctc ggcccaaagc atcagctcat cgagagcctg
    13621 cgcgacggac gcactgacgg tgtcgtccat cacagtttgc cagtgataca catggggatc
    13681 agcaatcgcg catatgaaat cacgccatgt agtgtattga ccgattcctt gcggtccgaa
    13741 tgggccgaac ccgctcgtct ggctaagatc ggccgcagcg atagcatcca tagcctccgc
    13801 gaccggctga agaacagcgg gcagttcggt ttcaggcagg tcttgcaacg tgacaccctg
    13861 tgcacggcgg gagatgcaat aggtcaggct ctcgctgaac tccccaatgt caagcacttc
    13921 cggaatcggg agcgcggccg atgcaaagtg ccgataaaca taacgatctt tgtagaaacc
    13981 atcggcgcag ctatttaccc gcaggacata tccacgccct cctacatcga agctgaaagc
    14041 acgagattct tcgccctccg agagctgcat caggtcggag acgctgtcga acttttcgat
    14101 cagaaacttc tcgacagacg tcgcggtgag ttcaggcttt ttcatagggg ggattcgagt
    14161 tgagagtgaa tatgagactc taattggata ccgaggggaa tttatggaac gtcagtggag
    14221 catttttgac aagaaatatt tgctagctga tagtgacctt aggcgacttt tgaacgcgca
    14281 ataatggttt ctgacgtatg tgcttagctc attaaactcc agaaacccgc ggctcagtgg
    14341 ctccttcaac gttgcggttc tgtcagttcc aaacgtaaaa cggcttgtcc cgcgtcatcg
    14401 gcgggggtca taacgtgact cccttaattc tccgctcatg atcgataatt c
//

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