pAtCas9-dead_1

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: pAtCas9-dead_1 
出品公司: 爱迪基因
目录编号: 91165
存储实验室: Daniel Voytas

载体骨架基本信息
载体名称: pCAMBIA
载体出品公司: --
载体抗性: 卡那霉素
空载体大小: --
完整质粒大小: --
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: AtCas9_D10A+H840A (dead)
物种来源: Synthetic
插入大小: --
基因突变: D10A, H840A
GenBank编号 --
启动子 --
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: pAtCas9-dead_1 was a gift from Daniel Voytas (Addgene plasmid # 91165 ; http://n2t.net/addgene:91165 ; RRID:Addgene_91165)
参考文献部分: A multi-purpose toolkit to enable advanced genome engineering in plants. Cermak T, Curtin SJ, Gil-Humanes J, Cegan R, Kono TJY, Konecna E, Belanto JJ, Starker CG, Mathre JW, Greenstein RL, Voytas DF. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. 10.1105/tpc.16.00922 PubMed 28522548

载体图谱质粒图谱和多克隆位点信息

pAtCas9-dead_1质粒图谱

载体简介

载体序列

LOCUS       pAtCas9-dead_1             14215 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  Cas9 only expression, Cas9 Type: AtCas9_D10A+H840A (dead), Plant 
            Selection: 2x35S:hpt II.
ACCESSION   .
VERSION     .
KEYWORDS    pAtCas9-dead_1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14215)
  AUTHORS   Cermak T, Curtin SJ, Gil-Humanes J, Cegan R, Kono TJY, Konecna E, 
            Belanto JJ, Starker CG, Mathre JW, Greenstein RL, Voytas DF
  TITLE     A multi-purpose toolkit to enable advanced genome engineering in 
            plants.
  JOURNAL   Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 
            10.1105/tpc.16.00922.
  PUBMED    28522548
REFERENCE   2  (bases 1 to 14215)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..14215
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(291..1085)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    1510..1534
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    1612..1786
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(1827..2852)
                     /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"
     promoter        complement(2919..3595)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     protein_bind    3786..3807
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3822..3852
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3860..3876
                     /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     3865..3887
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3884..3900
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3884..3900
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        4430..4775
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             4809..8912
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             8925..8945
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(9226..9243)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9226..9242)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9235..9257)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    9445..9469
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             10770..11399
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             11828..12901
                     /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      12967..13161
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(13397..13419)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    13505..13645
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     13560..13579
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(13660..13677)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13831..204)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13911..13930)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
       61 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
      121 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
      181 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
      241 cagtggaacg aaaactcacg ttaagggatt ttggtcatgc attctaggta ctaaaacaat
      301 tcatccagta aaatataata ttttattttc tcccaatcag gcttgatccc cagtaagtca
      361 aaaaatagct cgacatactg ttcttccccg atatcctccc tgatcgaccg gacgcagaag
      421 gcaatgtcat accacttgtc cgccctgccg cttctcccaa gatcaataaa gccacttact
      481 ttgccatctt tcacaaagat gttgctgtct cccaggtcgc cgtgggaaaa gacaagttcc
      541 tcttcgggct tttccgtctt taaaaaatca tacagctcgc gcggatcttt aaatggagtg
      601 tcttcttccc agttttcgca atccacatcg gccagatcgt tattcagtaa gtaatccaat
      661 tcggctaagc ggctgtctaa gctattcgta tagggacaat ccgatatgtc gatggagtga
      721 aagagcctga tgcactccgc atacagctcg ataatctttt cagggctttg ttcatcttca
      781 tactcttccg agcaaaggac gccatcggcc tcactcatga gcagattgct ccagccatca
      841 tgccgttcaa agtgcaggac ctttggaaca ggcagctttc cttccagcca tagcatcatg
      901 tccttttccc gttccacatc ataggtggtc cctttatacc ggctgtccgt catttttaaa
      961 tataggtttt cattttctcc caccagctta tataccttag caggagacat tccttccgta
     1021 tcttttacgc agcggtattt ttcgatcagt tttttcaatt ccggtgatat tctcatttta
     1081 gccatttatt atttccttcc tcttttctac agtatttaaa gataccccaa gaagctaatt
     1141 ataacaagac gaactccaat tcactgttcc ttgcattcta aaaccttaaa taccagaaaa
     1201 cagctttttc aaagttgttt tcaaagttgg cgtataacat agtatcgacg gagccgattt
     1261 tgaaaccgcg gtgatcacag gcagcaacgc tctgtcatcg ttacaatcaa catgctaccc
     1321 tccgcgagat catccgtgtt tcaaacccgg cagcttagtt gccgttcttc cgaatagcat
     1381 cggtaacatg agcaaagtct gccgccttac aacggctctc ccgctgacgc cgtcccggac
     1441 tgatgggctg cctgtatcga gtggtgattt tgtgccgagc tgccggtcgg ggagctgttg
     1501 gctggctggt ggcaggatat attgtggtgt aaacaaattg acgcttagac aacttaataa
     1561 cacattgcgg acgtttttaa tgtactgaat taacgccgaa ttaattcggg ggatctggat
     1621 tttagtactg gattttggtt ttaggaatta gaaattttat tgatagaagt attttacaaa
     1681 tacaaataca tactaagggt ttcttatatg ctcaacacat gagcgaaacc ctataggaac
     1741 cctaattccc ttatctggga actactcaca cattattatg gagaaactcg agcttgtcga
     1801 tcgacagatc ccggtcggca tctactctat ttctttgccc tcggacgagt gctggggcgt
     1861 cggtttccac tatcggcgag tacttctaca cagccatcgg tccagacggc cgcgcttctg
     1921 cgggcgattt gtgtacgccc gacagtcccg gctccggatc ggacgattgc gtcgcatcga
     1981 ccctgcgccc aagctgcatc atcgaaattg ccgtcaacca agctctgata gagttggtca
     2041 agaccaatgc ggagcatata cgcccggagt cgtggcgatc ctgcaagctc cggatgcctc
     2101 cgctcgaagt agcgcgtctg ctgctccata caagccaacc acggcctcca gaagaagatg
     2161 ttggcgacct cgtattggga atccccgaac atcgcctcgc tccagtcaat gaccgctgtt
     2221 atgcggccat tgtccgtcag gacattgttg gagccgaaat ccgcgtgcac gaggtgccgg
     2281 acttcggggc agtcctcggc ccaaagcatc agctcatcga gagcctgcgc gacggacgca
     2341 ctgacggtgt cgtccatcac agtttgccag tgatacacat ggggatcagc aatcgcgcat
     2401 atgaaatcac gccatgtagt gtattgaccg attccttgcg gtccgaatgg gccgaacccg
     2461 ctcgtctggc taagatcggc cgcagcgatc gcatccatag cctccgcgac cggttgtaga
     2521 acagcgggca gttcggtttc aggcaggtct tgcaacgtga caccctgtga acggcgggag
     2581 atgcaatagg tcaggctctc gctaaactcc ccaatgtcaa gcacttccgg aatcgggagc
     2641 gcggccgatg caaagtgccg ataaacataa cgatctttgt agaaaccatc ggcgcagcta
     2701 tttacccgca ggacatatcc acgccctcct acatcgaagc tgaaagcacg agattcttcg
     2761 ccctccgaga gctgcatcag gtcggagacg ctgtcgaact tttcgatcag aaacttctcg
     2821 acagacgtcg cggtgagttc aggctttttc atatctcatt gccccccgga tctgcgaaag
     2881 ctcgagagag atagatttgt agagagagac tggtgatttc agcgtgtcct ctccaaatga
     2941 aatgaacttc cttatataga ggaagggtct tgcgaaggat agtgggattg tgcgtcatcc
     3001 cttacgtcag tggagatatc acatcaatcc acttgctttg aagacgtggt tggaacgtct
     3061 tctttttcca cgatgctcct cgtgggtggg ggtccatctt tgggaccact gtcggcagag
     3121 gcatcttgaa cgatagcctt tcctttatcg caatgatggc atttgtaggt gccaccttcc
     3181 ttttctactg tccttttgat gaagtgacag atagctgggc aatggaatcc gaggaggttt
     3241 cccgatatta ccctttgttg aaaagtctca atagcccttt ggtcttctga gactgtatct
     3301 ttgatattct tggagtagac gagagtgtcg tgctccacca tgttcacatc aatccacttg
     3361 ctttgaagac gtggttggaa cgtcttcttt ttccacgatg ctcctcgtgg gtgggggtcc
     3421 atctttggga ccactgtcgg cagaggcatc ttgaacgata gcctttcctt tatcgcaatg
     3481 atggcatttg taggtgccac cttccttttc tactgtcctt ttgatgaagt gacagatagc
     3541 tgggcaatgg aatccgagga ggtttcccga tattaccctt tgttgaaaag tctcaatagc
     3601 cctttggtct tctgagactg tatctttgat attcttggag tagacgagag tgtcgtgctc
     3661 caccatgttg gcaagctgct ctagccaata cgcaaaccgc ctctccccgc gcgttggccg
     3721 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     3781 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     3841 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     3901 atgattacga attcgagctc ggtacccggg gatcggcgcg ccagatttgc cttttcaatt
     3961 tcagaaagaa tgctaaccca cagatggtta gagaggctta cgcagcaggt atcatcaaga
     4021 cgatctaccc gagcaataat ctccaggaaa tcaaatacct tcccaagaag gttaaagatg
     4081 cagtcaaaag attcaggact aactgcatca agaacacaga gaaagatata tttctcaaga
     4141 tcagaagtac tattccagta tggacgattc aaggcttgct tcacaaacca aggcaagtaa
     4201 tagagattgg agtctctaaa aaggtagttc ccactgaatc aaaggccatg gagtcaaaga
     4261 ttcaaataga ggacctaaca gaactcgccg taaagactgg cgaacagttc atacagagtc
     4321 tcttacgact caatgacaag aagaaaatct tcgtcaacat ggtggagcac gacacacttg
     4381 tctactccaa aaatatcaaa gatacagtct cagaagacca aagggcaatt gagacttttc
     4441 aacaaagggt aatatccgga aacctcctcg gattccattg cccagctatc tgtcacttta
     4501 ttgtgaagat agtggaaaag gaaggtggct cctacaaatg ccatcattgc gataaaggaa
     4561 aggccatcgt tgaagatgcc tctgccgaca gtggtcccaa agatggaccc ccacccacga
     4621 ggagcatcgt ggaaaaagaa gacgttccaa ccacgtcttc aaagcaagtg gattgatgtg
     4681 atatctccac tgacgtaagg gatgacgcac aatcccacta tccttcgcaa gacccttcct
     4741 ctatataagg aagttcattt catttggaga gaacacgggg gactcctgca ggtagatcgc
     4801 tcgtcgacat ggataagaag tactctatcg gactcgctat cggaactaac tctgtgggat
     4861 gggctgtgat caccgatgag tacaaggtgc catctaagaa gttcaaggtt ctcggaaaca
     4921 ccgataggca ctctatcaag aaaaacctta tcggtgctct cctcttcgat tctggtgaaa
     4981 ctgctgaggc taccagactc aagagaaccg ctagaagaag gtacaccaga agaaagaaca
     5041 ggatctgcta cctccaagag atcttctcta acgagatggc taaagtggat gattcattct
     5101 tccacaggct cgaagagtca ttcctcgtgg aagaagataa gaagcacgag aggcacccta
     5161 tcttcggaaa catcgttgat gaggtggcat accacgagaa gtaccctact atctaccacc
     5221 tcagaaagaa gctcgttgat tctactgata aggctgatct caggctcatc tacctcgctc
     5281 tcgctcacat gatcaagttc agaggacact tcctcatcga gggtgatctc aaccctgata
     5341 actctgatgt ggataagttg ttcatccagc tcgtgcagac ctacaaccag cttttcgaag
     5401 agaaccctat caacgcttca ggtgtggatg ctaaggctat cctctctgct aggctctcta
     5461 agtcaagaag gcttgagaac ctcattgctc agctccctgg tgagaagaag aacggacttt
     5521 tcggaaactt gatcgctctc tctctcggac tcacccctaa cttcaagtct aacttcgatc
     5581 tcgctgagga tgcaaagctc cagctctcaa aggataccta cgatgatgat ctcgataacc
     5641 tcctcgctca gatcggagat cagtacgctg atttgttcct cgctgctaag aacctctctg
     5701 atgctatcct cctcagtgat atcctcagag tgaacaccga gatcaccaag gctccactct
     5761 cagcttctat gatcaagaga tacgatgagc accaccagga tctcacactt ctcaaggctc
     5821 ttgttagaca gcagctccca gagaagtaca aagagatttt cttcgatcag tctaagaacg
     5881 gatacgctgg ttacatcgat ggtggtgcat ctcaagaaga gttctacaag ttcatcaagc
     5941 ctatcctcga gaagatggat ggaaccgagg aactcctcgt gaagctcaat agagaggatc
     6001 ttctcagaaa gcagaggacc ttcgataacg gatctatccc tcatcagatc cacctcggag
     6061 agttgcacgc tatccttaga aggcaagagg atttctaccc attcctcaag gataacaggg
     6121 aaaagattga gaagattctc accttcagaa tcccttacta cgtgggacct ctcgctagag
     6181 gaaactcaag attcgcttgg atgaccagaa agtctgagga aaccatcacc ccttggaact
     6241 tcgaagaggt ggtggataag ggtgctagtg ctcagtcttt catcgagagg atgaccaact
     6301 tcgataagaa ccttccaaac gagaaggtgc tccctaagca ctctttgctc tacgagtact
     6361 tcaccgtgta caacgagttg accaaggtta agtacgtgac cgagggaatg aggaagcctg
     6421 cttttttgtc aggtgagcaa aagaaggcta tcgttgatct cttgttcaag accaacagaa
     6481 aggtgaccgt gaagcagctc aaagaggatt acttcaagaa aatcgagtgc ttcgattcag
     6541 ttgagatttc tggtgttgag gataggttca acgcatctct cggaacctac cacgatctcc
     6601 tcaagatcat taaggataag gatttcttgg ataacgagga aaacgaggat atcttggagg
     6661 atatcgttct taccctcacc ctctttgaag atagagagat gattgaagaa aggctcaaga
     6721 cctacgctca tctcttcgat gataaggtga tgaagcagtt gaagagaaga agatacactg
     6781 gttggggaag gctctcaaga aagctcatta acggaatcag ggataagcag tctggaaaga
     6841 caatccttga tttcctcaag tctgatggat tcgctaacag aaacttcatg cagctcatcc
     6901 acgatgattc tctcaccttt aaagaggata tccagaaggc tcaggtttca ggacagggtg
     6961 atagtctcca tgagcatatc gctaacctcg ctggatctcc tgcaatcaag aagggaatcc
     7021 tccagactgt gaaggttgtg gatgagttgg tgaaggtgat gggaaggcat aagcctgaga
     7081 acatcgtgat cgaaatggct agagagaacc agaccactca gaagggacag aagaactcta
     7141 gggaaaggat gaagaggatc gaggaaggta tcaaagagct tggatctcag atcctcaaag
     7201 agcaccctgt tgagaacact cagctccaga atgagaagct ctacctctac tacctccaga
     7261 acggaaggga tatgtatgtg gatcaagagt tggatatcaa caggctctct gattacgatg
     7321 ttgatgctat cgtgccacag tcattcttga aggatgattc tatcgataac aaggtgctca
     7381 ccaggtctga taagaacagg ggtaagagtg ataacgtgcc aagtgaagag gttgtgaaga
     7441 aaatgaagaa ctattggagg cagctcctca acgctaagct catcactcag agaaagttcg
     7501 ataacttgac taaggctgag aggggaggac tctctgaatt ggataaggca ggattcatca
     7561 agaggcagct tgtggaaacc aggcagatca ctaagcacgt tgcacagatc ctcgattcta
     7621 ggatgaacac caagtacgat gagaacgata agttgatcag ggaagtgaag gttatcaccc
     7681 tcaagtcaaa gctcgtgtct gatttcagaa aggatttcca attctacaag gtgagggaaa
     7741 tcaacaacta ccaccacgct cacgatgctt accttaacgc tgttgttgga accgctctca
     7801 tcaagaagta tcctaagctc gagtcagagt tcgtgtacgg tgattacaag gtgtacgatg
     7861 tgaggaagat gatcgctaag tctgagcaag agatcggaaa ggctaccgct aagtatttct
     7921 tctactctaa catcatgaat ttcttcaaga ccgagattac cctcgctaac ggtgagatca
     7981 gaaagaggcc actcatcgag acaaacggtg aaacaggtga gatcgtgtgg gataagggaa
     8041 gggatttcgc taccgttaga aaggtgctct ctatgccaca ggtgaacatc gttaagaaaa
     8101 ccgaggtgca gaccggtgga ttctctaaag agtctatcct ccctaagagg aactctgata
     8161 agctcattgc taggaagaag gattgggacc ctaagaaata cggtggtttc gattctccta
     8221 ccgtggctta ctctgttctc gttgtggcta aggttgagaa gggaaagagt aagaagctca
     8281 agtctgttaa ggaacttctc ggaatcacta tcatggaaag gtcatctttc gagaagaacc
     8341 caatcgattt cctcgaggct aagggataca aagaggttaa gaaggatctc atcatcaagc
     8401 tcccaaagta ctcactcttc gaactcgaga acggtagaaa gaggatgctc gcttctgctg
     8461 gtgagcttca aaagggaaac gagcttgctc tcccatctaa gtacgttaac tttctttacc
     8521 tcgcttctca ctacgagaag ttgaagggat ctccagaaga taacgagcag aagcaacttt
     8581 tcgttgagca gcacaagcac tacttggatg agatcatcga gcagatctct gagttctcta
     8641 aaagggtgat cctcgctgat gcaaacctcg ataaggtgtt gtctgcttac aacaagcaca
     8701 gagataagcc tatcagggaa caggcagaga acatcatcca tctcttcacc cttaccaacc
     8761 tcggtgctcc tgctgctttc aagtacttcg atacaaccat cgataggaag agatacacct
     8821 ctaccaaaga agtgctcgat gctaccctca tccatcagtc tatcactgga ctctacgaga
     8881 ctaggatcga tctctcacag ctcggtggtg attcaagggc tgatcctaag aagaagagga
     8941 aggtttgact cgagatatga agatgaagat gaaatatttg gtgtgtcaaa taaaaagctt
     9001 gtgtgcttaa gtttgtgttt ttttcttggc ttgttgtgtt atgaatttgt ggctttttct
     9061 aatattaaat gaatgtaaga tcacattata atgaataaac aaatgtttct ataatccatt
     9121 gtgaatgttt tgttggatct cttctgcagc atataactac tgtatgtgct atggtatgga
     9181 ctatggaata tgattaaaga taaggagctc cggtgacgga ctggcactgg ccgtcgtttt
     9241 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     9301 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     9361 gcgcagcctg aatggcgaat gctagagcag cttgagcttg gatcagattg tcgtttcccg
     9421 ccttcagttt aaactatcag tgtttgacag gatatattgg cgggtaaacc taagagaaaa
     9481 gagcgtttat tagaataatc ggatatttaa aagggcgtga aaaggtttat ccgttcgtcc
     9541 atttgtatgt gcatgccaac cacagggttc ccctcgggat caaagtactt tgatccaacc
     9601 cctccgctgc tatagtgcag tcggcttctg acgttcagtg cagccgtctt ctgaaaacga
     9661 catgtcgcac aagtcctaag ttacgcgaca ggctgccgcc ctgccctttt cctggcgttt
     9721 tcttgtcgcg tgttttagtc gcataaagta gaatacttgc gactagaacc ggagacatta
     9781 cgccatgaac aagagcgccg ccgctggcct gctgggctat gcccgcgtca gcaccgacga
     9841 ccaggacttg accaaccaac gggccgaact gcacgcggcc ggctgcacca agctgttttc
     9901 cgagaagatc accggcacca ggcgcgaccg cccggagctg gccaggatgc ttgaccacct
     9961 acgccctggc gacgttgtga cagtgaccag gctagaccgc ctggcccgca gcacccgcga
    10021 cctactggac attgccgagc gcatccagga ggccggcgcg ggcctgcgta gcctggcaga
    10081 gccgtgggcc gacaccacca cgccggccgg ccgcatggtg ttgaccgtgt tcgccggcat
    10141 tgccgagttc gagcgttccc taatcatcga ccgcacccgg agcgggcgcg aggccgccaa
    10201 ggcccgaggc gtgaagtttg gcccccgccc taccctcacc ccggcacaga tcgcgcacgc
    10261 ccgcgagctg atcgaccagg aaggccgcac cgtgaaagag gcggctgcac tgcttggcgt
    10321 gcatcgctcg accctgtacc gcgcacttga gcgcagcgag gaagtgacgc ccaccgaggc
    10381 caggcggcgc ggtgccttcc gtgaggacgc attgaccgag gccgacgccc tggcggccgc
    10441 cgagaatgaa cgccaagagg aacaagcatg aaaccgcacc aggacggcca ggacgaaccg
    10501 tttttcatta ccgaagagat cgaggcggag atgatcgcgg ccgggtacgt gttcgagccg
    10561 cccgcgcacg tctcaaccgt gcggctgcat gaaatcctgg ccggtttgtc tgatgccaag
    10621 ctggcggcct ggccggccag cttggccgct gaagaaaccg agcgccgccg tctaaaaagg
    10681 tgatgtgtat ttgagtaaaa cagcttgcgt catgcggtcg ctgcgtatat gatgcgatga
    10741 gtaaataaac aaatacgcaa ggggaacgca tgaaggttat cgctgtactt aaccagaaag
    10801 gcgggtcagg caagacgacc atcgcaaccc atctagcccg cgccctgcaa ctcgccgggg
    10861 ccgatgttct gttagtcgat tccgatcccc agggcagtgc ccgcgattgg gcggccgtgc
    10921 gggaagatca accgctaacc gttgtcggca tcgaccgccc gacgattgac cgcgacgtga
    10981 aggccatcgg ccggcgcgac ttcgtagtga tcgacggagc gccccaggcg gcggacttgg
    11041 ctgtgtccgc gatcaaggca gccgacttcg tgctgattcc ggtgcagcca agcccttacg
    11101 acatatgggc caccgccgac ctggtggagc tggttaagca gcgcattgag gtcacggatg
    11161 gaaggctaca agcggccttt gtcgtgtcgc gggcgatcaa aggcacgcgc atcggcggtg
    11221 aggttgccga ggcgctggcc gggtacgagc tgcccattct tgagtcccgt atcacgcagc
    11281 gcgtgagcta cccaggcact gccgccgccg gcacaaccgt tcttgaatca gaacccgagg
    11341 gcgacgctgc ccgcgaggtc caggcgctgg ccgctgaaat taaatcaaaa ctcatttgag
    11401 ttaatgaggt aaagagaaaa tgagcaaaag cacaaacacg ctaagtgccg gccgtccgag
    11461 cgcacgcagc agcaaggctg caacgttggc cagcctggca gacacgccag ccatgaagcg
    11521 ggtcaacttt cagttgccgg cggaggatca caccaagctg aagatgtacg cggtacgcca
    11581 aggcaagacc attaccgagc tgctatctga atacatcgcg cagctaccag agtaaatgag
    11641 caaatgaata aatgagtaga tgaattttag cggctaaagg aggcggcatg gaaaatcaag
    11701 aacaaccagg caccgacgcc gtggaatgcc ccatgtgtgg aggaacgggc ggttggccag
    11761 gcgtaagcgg ctgggttgcc tgccggccct gcaatggcac tggaaccccc aagcccgagg
    11821 aatcggcgtg agcggtcgca aaccatccgg cccggtacaa atcggcgcgg cgctgggtga
    11881 tgacctggtg gagaagttga aggccgcgca ggccgcccag cggcaacgca tcgaggcaga
    11941 agcacgcccc ggtgaatcgt ggcaagcggc cgctgatcga atccgcaaag aatcccggca
    12001 accgccggca gccggtgcgc cgtcgattag gaagccgccc aagggcgacg agcaaccaga
    12061 ttttttcgtt ccgatgctct atgacgtggg cacccgcgat agtcgcagca tcatggacgt
    12121 ggccgttttc cgtctgtcga agcgtgaccg acgagctggc gaggtgatcc gctacgagct
    12181 tccagacggg cacgtagagg tttccgcagg gccggccggc atggccagtg tgtgggatta
    12241 cgacctggta ctgatggcgg tttcccatct aaccgaatcc atgaaccgat accgggaagg
    12301 gaagggagac aagcccggcc gcgtgttccg tccacacgtt gcggacgtac tcaagttctg
    12361 ccggcgagcc gatggcggaa agcagaaaga cgacctggta gaaacctgca ttcggttaaa
    12421 caccacgcac gttgccatgc agcgtacgaa gaaggccaag aacggccgcc tggtgacggt
    12481 atccgagggt gaagccttga ttagccgcta caagatcgta aagagcgaaa ccgggcggcc
    12541 ggagtacatc gagatcgagc tagctgattg gatgtaccgc gagatcacag aaggcaagaa
    12601 cccggacgtg ctgacggttc accccgatta ctttttgatc gatcccggca tcggccgttt
    12661 tctctaccgc ctggcacgcc gcgccgcagg caaggcagaa gccagatggt tgttcaagac
    12721 gatctacgaa cgcagtggca gcgccggaga gttcaagaag ttctgtttca ccgtgcgcaa
    12781 gctgatcggg tcaaatgacc tgccggagta cgatttgaag gaggaggcgg ggcaggctgg
    12841 cccgatccta gtcatgcgct accgcaacct gatcgagggc gaagcatccg ccggttccta
    12901 atgtacggag cagatgctag ggcaaattgc cctagcaggg gaaaaaggtc gaaaaggtct
    12961 ctttcctgtg gatagcacgt acattgggaa cccaaagccg tacattggga accggaaccc
    13021 gtacattggg aacccaaagc cgtacattgg gaaccggtca cacatgtaag tgactgatat
    13081 aaaagagaaa aaaggcgatt tttccgccta aaactcttta aaacttatta aaactcttaa
    13141 aacccgcctg gcctgtgcat aactgtctgg ccagcgcaca gccgaagagc tgcaaaaagc
    13201 gcctaccctt cggtcgctgc gctccctacg ccccgccgct tcgcgtcggc ctatcgcggc
    13261 cgctggccgc tcaaaaatgg ctggcctacg gccaggcaat ctaccagggc gcggacaagc
    13321 cgcgccgtcg ccactcgacc gccggcgccc acatcaaggc accctgcctc gcgcgtttcg
    13381 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
    13441 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
    13501 ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc
    13561 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
    13621 cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg
    13681 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
    13741 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
    13801 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
    13861 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
    13921 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
    13981 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
    14041 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
    14101 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
    14161 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtat
//

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