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pTCK303-Stuffer 植物干扰shRNA

pTCK303-Stuffer 植物干扰shRNA
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  • 产品名称:pTCK303-Stuffer 植物干扰shRNA
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  • 产品展商:XYbscience
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简单介绍
pTCK303-Stuffer 植物干扰shRNA菌株发货前���经过严格的多重验证,如存在质量问题,请在收到产品的三个月内通知我司。pTCK303-Stuffer 植物干扰shRNA收到质粒后请短暂离心,加入20μl ddH2O溶解质粒,取2μl转化至对应感受态中,挑取单克隆重新提取质粒后使用。
产品描述

pTCK303-Stuffer 植物干扰shRNA

  • 产品信息
  • 基本信息
  • 质粒简介
  • 质粒图谱
  • 质粒序列

产品信息

产品货号 产品名称 产品规格 优惠价
XY0273 pTCK303-Stuffer 植物干扰shRNA

2μg质粒

¥请加QQ询价

使用说明

信裕质粒平台的各批次质粒菌株发货前均经过严格的多重验证,如存在质量问题,请在收到产品的三个月内通知我司。收到质粒后请短暂离心,加入20μl ddH2O溶解质粒,取2μl转化至对应感受态中,挑取单克隆重新提取质粒后使用。如需获取其他详细信息请登录我司官网查询。

基本信息

启动子: Ubi
复制子: ori
终止子: NOS
质粒分类: 植物系列,RNAi载体
原核抗性: 卡那霉素Kan
筛选标记: 潮霉素HygR
克隆菌株: 大肠杆菌DH5α
培养条件: 37℃,有氧,LB
表达宿主: 植物细胞

质粒简介

          RNA interferences (RNAi) has proven to be an effective strategy to knock out homologous genes in a wide range of species. Based on its principle, a new generation of vectors containing an inverted target sequence separated by an intron as a loop, developing simplifications to the procedure of RNAi construction are required to improve the efficiency of gene inactivation techniques. Here, a novel polymerase chain reaction (PCR)—based RNAi vector pTCK303 with a maize ubiquitin promoter, 2 specific multiple enzyme sites, and a rice intron was constructed for monocot gene silencing. With this vector, only 1 PCR product amplified by a single pair of primers and 2 ligation reactions were needed to create an RNAi construct, which shortened the time span before being transformed into the plant. To test the efficiency of vector pTCK303, a rice geneOsGAS1 was used, and its RNAi construct was introduced into rice calli. Southern blot analysis of the transgenic rice confirmed the presence of theOsGAS1 RNAi structure. The decrease inOsGAS1 level in the transgenic rice was detected by Northern blot probed with anOsGAS1-specific sequence. Moreover, the rate of inhibition of the RNA expression level in RNAi transgenic rice was approximately 85% according to our real-time PCR. Therefore, the RNAi vector pTCK303 based on the homology-dependent gene-silencing mechanisms facilitated the inhibition of endogenous genes in a monocot and was proven to be a practical and efficient platform for silencing a rice gene.

质粒图谱

质粒序列

LOCUS       Exported               14856 bp ds-DNA     circular SYN 15-JAN-2018
DEFINITION  synthetic circular DNA
FEATURES             Location/Qualifiers
     source          1..14856
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             join(1..9,200..2017)
                     /codon_start=1
                     /product="beta-glucuronidase"
                     /label=GUS
                     /note="This version of the gusA gene has a 5' extension 
                     with a catalase intron to ensure expression in plants but 
                     not bacteria."
                     /translation="DLRNRRLVRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESAL
                     QESRAIAVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVW
                     VNNQEVMEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKK
                     QSYFHDFFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVEL
                     RDADQQVVATGQGTSGTLQVVNPHLWQPGEGYLYELEVTAKSQTESDIYPLRVGIRSVA
                     VKGQQFLINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYA
                     EEMLDWADEHGIVVIDETAAVGFQLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIK
                     ELIARDKNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAH
                     TDTISDLFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLA
                     GLHSMYTDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGI
                     FTRDRKPKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     intron          10..199
                     /label=cat1 intron
                     /note="castor bean catalase intron, modified"
     CDS             2024..2041
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2076..2328
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    2350..2374
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3674..4303
                     /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    6408..6548
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(6734..7322)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7409..8203)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    8628..8652
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    8730..8904
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(8944..9969)
                     /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
                     K"
     promoter        complement(10036..10713)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     terminator      11036..11288
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    11305..11546
                     /label=Stuffer
     misc_feature    11553..12060
                     /label=Rice intron
     promoter        complement(12088..14080)
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     promoter        14489..14834
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
ORIGIN
        1 gatctgaggg taaatttcta gtttttctcc ttcattttct tggttaggac ccttttctct
       61 ttttattttt ttgagctttg atctttcttt aaactgatct attttttaat tgattggtta
      121 tggtgtaaat attacatagc tttaactgat aatctgatta ctttatttcg tgtgtctatg
      181 atgatgatga tagttacaga accgacgact cgtccgtcct gtagaaaccc caacccgtga
      241 aatcaaaaaa ctcgacggcc tgtgggcatt cagtctggat cgcgaaaact gtggaattga
      301 tcagcgttgg tgggaaagcg cgttacaaga aagccgggca attgctgtgc caggcagttt
      361 taacgatcag ttcgccgatg cagatattcg taattatgcg ggcaacgtct ggtatcagcg
      421 cgaagtcttt ataccgaaag gttgggcagg ccagcgtatc gtgctgcgtt tcgatgcggt
      481 cactcattac ggcaaagtgt gggtcaataa tcaggaagtg atggagcatc agggcggcta
      541 tacgccattt gaagccgatg tcacgccgta tgttattgcc gggaaaagtg tacgtatcac
      601 cgtttgtgtg aacaacgaac tgaactggca gactatcccg ccgggaatgg tgattaccga
      661 cgaaaacggc aagaaaaagc agtcttactt ccatgatttc tttaactatg ccggaatcca
      721 tcgcagcgta atgctctaca ccacgccgaa cacctgggtg gacgatatca ccgtggtgac
      781 gcatgtcgcg caagactgta accacgcgtc tgttgactgg caggtggtgg ccaatggtga
      841 tgtcagcgtt gaactgcgtg atgcggatca acaggtggtt gcaactggac aaggcactag
      901 cgggactttg caagtggtga atccgcacct ctggcaaccg ggtgaaggtt atctctatga
      961 actcgaagtc acagccaaaa gccagacaga gtctgatatc tacccgcttc gcgtcggcat
     1021 ccggtcagtg gcagtgaagg gccaacagtt cctgattaac cacaaaccgt tctactttac
     1081 tggctttggt cgtcatgaag atgcggactt acgtggcaaa ggattcgata acgtgctgat
     1141 ggtgcacgac cacgcattaa tggactggat tggggccaac tcctaccgta cctcgcatta
     1201 cccttacgct gaagagatgc tcgactgggc agatgaacat ggcatcgtgg tgattgatga
     1261 aactgctgct gtcggctttc agctgtcttt aggcattggt ttcgaagcgg gcaacaagcc
     1321 gaaagaactg tacagcgaag aggcagtcaa cggggaaact cagcaagcgc acttacaggc
     1381 gattaaagag ctgatagcgc gtgacaaaaa ccacccaagc gtggtgatgt ggagtattgc
     1441 caacgaaccg gatacccgtc cgcaaggtgc acgggaatat ttcgcgccac tggcggaagc
     1501 aacgcgtaaa ctcgacccga cgcgtccgat cacctgcgtc aatgtaatgt tctgcgacgc
     1561 tcacaccgat accatcagcg atctctttga tgtgctgtgc ctgaaccgtt attacggatg
     1621 gtatgtccaa agcggcgatt tggaaacggc agagaaggta ctggaaaaag aacttctggc
     1681 ctggcaggag aaactgcatc agccgattat catcaccgaa tacggcgtgg atacgttagc
     1741 cgggctgcac tcaatgtaca ccgacatgtg gagtgaagag tatcagtgtg catggctgga
     1801 tatgtatcac cgcgtctttg atcgcgtcag cgccgtcgtc ggtgaacagg tatggaattt
     1861 cgccgatttt gcgacctcgc aaggcatatt gcgcgttggc ggtaacaaga aagggatctt
     1921 cactcgcgac cgcaaaccga agtcggcggc ttttctgctg caaaaacgct ggactggcat
     1981 gaacttcggt gaaaaaccgc agcagggagg caaacaagct agccaccacc accaccacca
     2041 cgtgtgaatt acaggtgacc agctcgaatt tccccgatcg ttcaaacatt tggcaataaa
     2101 gtttcttaag attgaatcct gttgccggtc ttgcgatgat tatcatataa tttctgttga
     2161 attacgttaa gcatgtaata attaacatgt aatgcatgac gttatttatg agatgggttt
     2221 ttatgattag agtcccgcaa ttatacattt aatacgcgat agaaaacaaa atatagcgcg
     2281 caaactagga taaattatcg cgcgcggtgt catctatgtt actagatcgg gaattaaact
     2341 atcagtgttt gacaggatat attggcgggt aaacctaaga gaaaagagcg tttattagaa
     2401 taacggatat ttaaaagggc gtgaaaaggt ttatccgttc gtccatttgt atgtgcatgc
     2461 caaccacagg gttcccctcg ggatcaaagt actttgatcc aacccctccg ctgctatagt
     2521 gcagtcggct tctgacgttc agtgcagccg tcttctgaaa acgacatgtc gcacaagtcc
     2581 taagttacgc gacaggctgc cgccctgccc ttttcctggc gttttcttgt cgcgtgtttt
     2641 agtcgcataa agtagaatac ttgcgactag aaccggagac attacgccat gaacaagagc
     2701 gccgccgctg gcctgctggg ctatgcccgc gtcagcaccg acgaccagga cttgaccaac
     2761 caacgggccg aactgcacgc ggccggctgc accaagctgt tttccgagaa gatcaccggc
     2821 accaggcgcg accgcccgga gctggccagg atgcttgacc acctacgccc tggcgacgtt
     2881 gtgacagtga ccaggctaga ccgcctggcc cgcagcaccc gcgacctact ggacattgcc
     2941 gagcgcatcc aggaggccgg cgcgggcctg cgtagcctgg cagagccgtg ggccgacacc
     3001 accacgccgg ccggccgcat ggtgttgacc gtgttcgccg gcattgccga gttcgagcgt
     3061 tccctaatca tcgaccgcac ccggagcggg cgcgaggccg ccaaggcccg aggcgtgaag
     3121 tttggccccc gccctaccct caccccggca cagatcgcgc acgcccgcga gctgatcgac
     3181 caggaaggcc gcaccgtgaa agaggcggct gcactgcttg gcgtgcatcg ctcgaccctg
     3241 taccgcgcac ttgagcgcag cgaggaagtg acgcccaccg aggccaggcg gcgcggtgcc
     3301 ttccgtgagg acgcattgac cgaggccgac gccctggcgg ccgccgagaa tgaacgccaa
     3361 gaggaacaag catgaaaccg caccaggacg gccaggacga accgtttttc attaccgaag
     3421 agatcgaggc ggagatgatc gcggccgggt acgtgttcga gccgcccgcg cacgtctcaa
     3481 ccgtgcggct gcatgaaatc ctggccggtt tgtctgatgc caagctggcg gcctggccgg
     3541 ccagcttggc cgctgaagaa accgagcgcc gccgtctaaa aaggtgatgt gtatttgagt
     3601 aaaacagctt gcgtcatgcg gtcgctgcgt atatgatgcg atgagtaaat aaacaaatac
     3661 gcaaggggaa cgcatgaagg ttatcgctgt acttaaccag aaaggcgggt caggcaagac
     3721 gaccatcgca acccatctag cccgcgccct gcaactcgcc ggggccgatg ttctgttagt
     3781 cgattccgat ccccagggca gtgcccgcga ttgggcggcc gtgcgggaag atcaaccgct
     3841 aaccgttgtc ggcatcgacc gcccgacgat tgaccgcgac gtgaaggcca tcggccggcg
     3901 cgacttcgta gtgatcgacg gagcgcccca ggcggcggac ttggctgtgt ccgcgatcaa
     3961 ggcagccgac ttcgtgctga ttccggtgca gccaagccct tacgacatat gggccaccgc
     4021 cgacctggtg gagctggtta agcagcgcat tgaggtcacg gatggaaggc tacaagcggc
     4081 ctttgtcgtg tcgcgggcga tcaaaggcac gcgcatcggc ggtgaggttg ccgaggcgct
     4141 ggccgggtac gagctgccca ttcttgagtc ccgtatcacg cagcgcgtga gctacccagg
     4201 cactgccgcc gccggcacaa ccgttcttga atcagaaccc gagggcgacg ctgcccgcga
     4261 ggtccaggcg ctggccgctg aaattaaatc aaaactcatt tgagttaatg aggtaaagag
     4321 aaaatgagca aaagcacaaa cacgctaagt gccggccgtc cgagcgcacg cagcagcaag
     4381 gctgcaacgt tggccagcct ggcagacacg ccagccatga agcgggtcaa ctttcagttg
     4441 ccggcggagg atcacaccaa gctgaagatg tacgcggtac gccaaggcaa gaccattacc
     4501 gagctgctat ctgaatacat cgcgcagcta ccagagtaaa tgagcaaatg aataaatgag
     4561 tagatgaatt ttagcggcta aaggaggcgg catggaaaat caagaacaac caggcaccga
     4621 cgccgtggaa tgccccatgt gtggaggaac gggcggttgg ccaggcgtaa gcggctgggt
     4681 tgtctgccgg ccctgcaatg gcactggaac ccccaagccc gaggaatcgg cgtgacggtc
     4741 gcaaaccatc cggcccggta caaatcggcg cggcgctggg tgatgacctg gtggagaagt
     4801 tgaaggccgc gcaggccgcc cagcggcaac gcatcgaggc agaagcacgc cccggtgaat
     4861 cgtggcaagc ggccgctgat cgaatccgca aagaatcccg gcaaccgccg gcagccggtg
     4921 cgccgtcgat taggaagccg cccaagggcg acgagcaacc agattttttc gttccgatgc
     4981 tctatgacgt gggcacccgc gatagtcgca gcatcatgga cgtggccagt tttaaagagt
     5041 tttaggcgga aaaatcgcct tttttctctt ttatatcagt cacttacatg tgtgaccggt
     5101 tcccaatgta cggctttggg ttcccaatgt acgggttccg gttcccaatg tacggctttg
     5161 ggttcccaat gtacgtgcta tccacaggaa agagaccttt tcgacctttt tcccctgcta
     5221 gggcaatttg ccctagcatc tgctccgtac attaggaacc ggcggatgct tcgccctcga
     5281 tcaggttgcg gtagcgcatg actaggatcg ggccagcctg ccccgcctcc tccttcaaat
     5341 cgtactccgg caggtcattt gacccgatca gcttgcgcac ggtgaaacag aacttcttga
     5401 actctccggc gctgccactg cgttcgtaga tcgtcttgaa caaccatctg gcttctgcct
     5461 tgcctgcggc gcggcgtgcc aggcggtaga gaaaacggcc gatgccggga tcgatcaaaa
     5521 agtaatcggg gtgaaccgtc agcacgtccg ggttcttgcc ttctgtgatc tcgcggtaca
     5581 tccaatcagc tagctcgatc tcgatgtact ccggccgccc ggtttcgctc tttacgatct
     5641 tgtagcggct aatcaaggct tcaccctcgg ataccgtcac caggcggccg ttcttggcct
     5701 tcttcgtacg ctgcatggca acgtgcgtgg tgtttaaccg aatgcaggtt tctaccaggt
     5761 cgtctttctg ctttccgcca tcggctcgcc ggcagaactt gagtacgtcc gcaacgtgtg
     5821 gacggaacac gcggccgggc ttgtctccct tcccttcccg gtatcggttc atggattcgg
     5881 ttagatggga aaccgccatc agtaccaggt cgtaatccca cacactggcc atgccggccg
     5941 gccctgcgga aacctctacg tgcccgtctg gaagctcgta gcggatcacc tcgccagctc
     6001 gtcggtcacg cttcgacaga cggaaaacta ttaaaactct taaaacccgc ctggcctgtg
     6061 cataactgtc tggccagcgc acagccgaag agctgcaaaa agcgcctacc cttcggtcgc
     6121 tgcgctccct acgccccgcc gcttcgcgtc ggcctatcgc ggccgctggc cgctcaaaaa
     6181 tggctggcct acggccaggc aatctaccag ggcgcggaca agccgcgccg tcgccactcg
     6241 accgccggcg cccacatcaa ggcaccctgc ctcgcgcgtt tcggtgatga cggtgaaaac
     6301 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     6361 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc
     6421 cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg
     6481 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     6541 gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6601 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6661 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6721 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6781 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6841 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6901 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     6961 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     7021 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     7081 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7141 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     7201 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7261 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7321 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7381 aagggatttt ggtcatgcat tctaggtact aaaacaattc atccagtaaa atataatatt
     7441 ttattttctc ccaatcaggc ttgatcccca gtaagtcaaa aaatagctcg acatactgtt
     7501 cttccccgat atcctccctg atcgaccgga cgcagaaggc aatgtcatac cacttgtccg
     7561 ccctgccgct tctcccaaga tcaataaagc cacttacttt gccatctttc acaaagatgt
     7621 tgctgtctcc caggtcgccg tgggaaaaga caagttcctc ttcgggcttt tccgtcttta
     7681 aaaaatcata cagctcgcgc ggatctttaa atggagtgtc ttcttcccag ttttcgcaat
     7741 ccacatcggc cagatcgtta ttcagtaagt aatccaattc ggctaagcgg ctgtctaagc
     7801 tattcgtata gggacaatcc gatatgtcga tggagtgaaa gagcctgatg cactccgcat
     7861 acagctcgat aatcttttca gggctttgtt catcttcata ctcttccgag caaaggacgc
     7921 catcggcctc actcatgagc agattgctcc agccatcatg ccgttcaaag tgcaggacct
     7981 ttggaacagg cagctttcct tccagccata gcatcatgtc cttttcccgt tccacatcat
     8041 aggtggtccc tttataccgg ctgtccgtca tttttaaata taggttttca ttttctccca
     8101 ccagcttata taccttagca ggagacattc cttccgtatc ttttacgcag cggtattttt
     8161 cgatcagttt tttcaattcc ggtgatattc tcattttagc catttattat ttccttcctc
     8221 ttttctacag tatttaaaga taccccaaga agctaattat aacaagacga actccaattc
     8281 actgttcctt gcattctaaa accttaaata ccagaaaaca gctttttcaa agttgttttc
     8341 aaagttggcg tataacatag tatcgacgga gccgattttg aaaccgcggt gatcacaggc
     8401 agcaacgctc tgtcatcgtt acaatcaaca tgctaccctc cgcgagatca tccgtgtttc
     8461 aaacccggca gcttagttgc cgttcttccg aatagcatcg gtaacatgag caaagtctgc
     8521 cgccttacaa cggctctccc gctgacgccg tcccggactg atgggctgcc tgtatcgagt
     8581 ggtgattttg tgccgagctg ccggtcgggg agctgttggc tggctggtgg caggatatat
     8641 tgtggtgtaa acaaattgac gcttagacaa cttaataaca cattgcggac gtttttaatg
     8701 tactgaatta acgccgaatt aattcggggg atctggattt tagtactgga ttttggtttt
     8761 aggaattaga aattttattg atagaagtat tttacaaata caaatacata ctaagggttt
     8821 cttatatgct caacacatga gcgaaaccct ataggaaccc taattccctt atctgggaac
     8881 tactcacaca ttattatgga gaaactcgag cttgtcgatc gacagatccg gtcggcatct
     8941 actctatttc tttgccctcg gacgagtgct ggggcgtcgg tttccactat cggcgagtac
     9001 ttctacacag ccatcggtcc agacggccgc gcttctgcgg gcgatttgtg tacgcccgac
     9061 agtcccggct ccggatcgga cgattgcgtc gcatcgaccc tgcgcccaag ctgcatcatc
     9121 gaaattgccg tcaaccaagc tctgatagag ttggtcaaga ccaatgcgga gcatatacgc
     9181 ccggagtcgt ggcgatcctg caagctccgg atgcctccgc tcgaagtagc gcgtctgctg
     9241 ctccatacaa gccaaccacg gcctccagaa gaagatgttg gcgacctcgt attgggaatc
     9301 cccgaacatc gcctcgctcc agtcaatgac cgctgttatg cggccattgt ccgtcaggac
     9361 attgttggag ccgaaatccg cgtgcacgag gtgccggact tcggggcagt cctcggccca
     9421 aagcatcagc tcatcgagag cctgcgcgac ggacgcactg acggtgtcgt ccatcacagt
     9481 ttgccagtga tacacatggg gatcagcaat cgcgcatatg aaatcacgcc atgtagtgta
     9541 ttgaccgatt ccttgcggtc cgaatgggcc gaacccgctc gtctggctaa gatcggccgc
     9601 agcgatcgca tccatagcct ccgcgaccgg ttgtagaaca gcgggcagtt cggtttcagg
     9661 caggtcttgc aacgtgacac cctgtgcacg gcgggagatg caataggtca ggctctcgct
     9721 aaactcccca atgtcaagca cttccggaat cgggagcgcg gccgatgcaa agtgccgata
     9781 aacataacga tctttgtaga aaccatcggc gcagctattt acccgcagga catatccacg
     9841 ccctcctaca tcgaagctga aagcacgaga ttcttcgccc tccgagagct gcatcaggtc
     9901 ggagacgctg tcgaactttt cgatcagaaa cttctcgaca gacgtcgcgg tgagttcagg
     9961 ctttttcata tctcattgcc ccccgggatc tgcgaaagct cgaggagata gatttgtaga
    10021 gagagactgg tgatttcagc gtgtcctctc caaatgaaat gaacttcctt atatagagga
    10081 aggtcttgcg aaggatagtg ggattgtgcg tcatccctta cgtcagtgga gatatcacat
    10141 caatccactt gctttgaaga cgtggttgga acgtcttctt tttccacgat gctcctcgtg
    10201 ggtgggggtc catctttggg accactgtcg gcagaggcat cttgaacgat agcctttcct
    10261 ttatcgcaat gatggcattt gtaggtgcca ccttcctttt ctactgtcct tttgatgaag
    10321 tgacagatag ctgggcaatg gaatccgagg aggtttcccg atattaccct ttgttgaaaa
    10381 gtctcaatag ccctttggtc ttctgagact gtatctttga tattcttgga gtagacgaga
    10441 gtgtcgtgct ccaccatgtt atcacatcaa tccacttgct ttgaagacgt ggttggaacg
    10501 tcttcttttt ccacgatgct cctcgtgggt gggggtccat ctttgggacc actgtcggca
    10561 gaggcatctt gaacgatagc ctttccttta tcgcaatgat ggcatttgta ggtgccacct
    10621 tccttttcta ctgtcctttt gatgaagtga cagatagctg ggcaatggaa tccgaggagg
    10681 tttcccgata ttaccctttg ttgaaaagtc tcaatagccc tttggtcttc tgagactgta
    10741 tctttgatat tcttggagta gacgagagtg tcgtgctcca ccatgtttgg caagctgctc
    10801 tagccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
    10861 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
    10921 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
    10981 gtgagcggat aacaatttca cacaggaaac agctatgaca tgattacgaa ttcccgatct
    11041 agtaacatag atgacaccgc gcgcgataat ttatcctagt ttgcgcgcta tattttgttt
    11101 tctatcgcgt attaaatgta taattgcggg actctaatca taaaaaccca tctcataaat
    11161 aacgtcatgc attacatgtt aattattaca tgcttaacgt aattcaacag aaattatatg
    11221 ataatcatcg caagaccggc aacaggattc aatcttaaga aactttattg ccaaatgttt
    11281 gaacgatcgg ggaaattcga gctcattaca ttgatgagaa atattacgta catatacata
    11341 catccacgtc gatgcttaga agttacagca tgaaccaggg tttttattga acatgtcgaa
    11401 agcgtttata aaacataaac acacatttaa ttacggcaca ttcccaccgg ccgcctatat
    11461 attgatcgcg ccggtcaagt attttttttc ccaccttttc agcggacgcc gggtcgatta
    11521 gatggggatc gcgctggtga tccgcgacta gtatcgattg aaaatctcga aacagccgtg
    11581 tcatagtcaa tcattaggtg ttataggaac aatcaaaggt tttttcaagt gttaatcttc
    11641 atactaatat atacagtggg tactctttat ctactgccgt ggaactgtca tatttgatta
    11701 tgaaatttta gctctagaaa atatttgatc atcaatgtca agactttatg accttgcaaa
    11761 atacatttcc taattgagaa cagggtaaaa ttatgaacta tgcctctgaa ccttcataca
    11821 caggcagcac attttttgtt gtaaaattca tcttaatatc agcggaaaga ctggaccaga
    11881 gaaagaaaaa gttaagacag gcatatactc ttgatcctct aaaagagatg aggcggtaca
    11941 atgatcaacc atgaacatta aagtgatacg tggaacatga gaacacaaat aattgtcact
    12001 ggaacataag tacaaaaagg tttgtagtaa cttaccgcta gcagatctgt cgacctcgag
    12061 ggtacccggg gatcctctag agtcgacctg cagaagtaac accaaacaac agggtgagca
    12121 tcgacaaaag aaacagtacc aagcaaataa atagcgtatg aaggcagggc taaaaaaatc
    12181 cacatatagc tgctgcatat gccatcatcc aagtatatca agatcaaaat aattataaaa
    12241 catacttgtt tattataata gataggtact caaggttaga gcatatgaat agatgctgca
    12301 tatgccatca tgtatatgca tcagtaaaac ccacatcaac atgtatacct atcctagatc
    12361 gatatttcca tccatcttaa actcgtaact atgaagatgt atgacacaca catacagttc
    12421 caaaattaat aaatacacca ggtagtttga aacagtattc tactccgatc tagaacgaat
    12481 gaacgaccgc ccaaccacac cacatcatca caaccaagcg aacaaaaagc atctctgtat
    12541 atgcatcagt aaaacccgca tcaacatgta tacctatcct agatcgatat ttccatccat
    12601 catcttcaat tcgtaactat gaatatgtat ggcacacaca tacagatcca aaattaataa
    12661 atccaccagg tagtttgaaa cagaattcta ctccgatcta gaacgaccgc ccaaccagac
    12721 cacatcatca caaccaagac aaaaaaaagc atgaaaagat gacccgacaa acaagtgcac
    12781 ggcatatatt gaaataaagg aaaagggcaa accaaaccct atgcaacgaa acaaaaaaaa
    12841 tcatgaaatc gatcccgtct gcggaacggc tagagccatc ccaggattcc ccaaagagaa
    12901 acactggcaa gttagcaatc agaacgtgtc tgacgtacag gtcgcatccg tgtacgaacg
    12961 ctagcagcac ggatctaaca caaacacgga tctaacacaa acatgaacag aagtagaact
    13021 accgggccct aaccatggac cggaacgccg atctagagaa ggtagagagg gggggggggg
    13081 gaggacgagc ggcgtacctt gaagcggagg tgccgacggg tggatttggg ggagatctgg
    13141 ttgtgtgtgt gtgcgctccg aacaacacga ggttggggaa agagggtgtg gagggggtgt
    13201 ctatttatta cggcgggcga ggaagggaaa gcgaaggagc ggtgggaaag gaatcccccg
    13261 tagctgccgg tgccgtgaga ggaggaggag gccgcctgcc gtgccggctc acgtctgccg
    13321 ctccgccacg caatttctgg atgccgacag cggagcaagt ccaacggtgg agcggaactc
    13381 tcgagagggg tccagaggca gcgacagaga tgccgtgccg tctgcttcgc ttggcccgac
    13441 gcgacgctgc tggttcgctg gttggtgtcc gttagactcg tcgacggcgt ttaacaggct
    13501 ggcattatct actcgaaaca agaaaaatgt ttccttagtt tttttaattt cttaaagggt
    13561 atttgtttaa tttttagtca ctttatttta ttctatttta tatctaaatt attaaataaa
    13621 aaaactaaaa tagagtttta gttttcttaa tttagaggct aaaatagaat aaaatagatg
    13681 tactaaaaaa attagtctat aaaaaccatt aaccctaaac cctaaatgga tgtactaata
    13741 aaatggatga agtattatat aggtgaagct atttgcaaaa aaaaaggaga acacatgcac
    13801 actaaaaaga taaaactgta gagtcctgtt gtcaaaatac tcaattgtcc tttagaccat
    13861 gtctaactgt tcatttatat gattctctaa aacactgata ttattgtagt actatagatt
    13921 atattattcg tagagtaaag tttaaatata tgtataaaga tagataaact gcacttcaaa
    13981 caagtgtgac aaaaaaaata tgtggtaatt ttttataact tagacatgca atgctcatta
    14041 tctctagaga ggggcacgac cgggtcacgc tgcactgcag gcatgcaagc ttggcactgg
    14101 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
    14161 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
    14221 cccaacagtt gcgcagcctg aatggcgaat gctagagcag cttgagcttg gatcagattg
    14281 tcgtttcccg ccttcagttt agcttcatgg agtcaaagat tcaaatagag gacctaacag
    14341 aactcgccgt aaagactggc gaacagttca tacagagtct cttacgactc aatgacaaga
    14401 agaaaatctt cgtcaacatg gtggagcacg acacacttgt ctactccaaa aatatcaaag
    14461 atacagtctc agaagaccaa agggcaattg agacttttca acaaagggta atatccggaa
    14521 acctcctcgg attccattgc ccagctatct gtcactttat tgtgaagata gtggaaaagg
    14581 aaggtggctc ctacaaatgc catcattgcg ataaaggaaa ggccatcgtt gaagatgcct
    14641 ctgccgacag tggtcccaaa gatggacccc cacccacgag gagcatcgtg gaaaaagaag
    14701 acgttccaac cacgtcttca aagcaagtgg attgatgtga tatctccact gacgtaaggg
    14761 atgacgcaca atcccactat ccttcgcaag acccttcctc tatataagga agttcatttc
    14821 atttggagag aacacggggg actcttgacc atggta
//
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