pCDH-ECMV-MCS-3×FLAG-EF1α-ZsGreen1-T2A-Puro

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产品名称：pCDH-ECMV-MCS-3×FLAG-EF1α-ZsGreen1-T2A-Puro
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简单介绍

pCDH-ECMV-MCS-3×FLAG-EF1α-ZsGreen1-T2A-Puro质粒是在pCDH载体的基础上改造而成，CMV启动目的基因表达，C端的3×FLAG融合标签能很好的被FLAG抗体识别。pCDH-ECMV-MCS-3×FLAG-EF1α-ZsGreen1-T2A-Puro该质粒可作为过表达质粒直接转染哺乳细胞，也可包装成慢病毒侵染细胞。质粒本身带有ZsGreen1荧光蛋白和Puro筛选标记，可利用荧光显微镜观察转染效率，也可用嘌呤霉素筛选转染后的细胞得到阳性克隆。

产品描述

pCDH-ECMV-MCS-3×FLAG-EF1α-ZsGreen1-T2A-Puro

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
XY1307	pCDH-ECMV-MCS-3×FLAG-EF1α-ZsGreen1-T2A-Puro	5μg	¥请询价

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:	ECMV promoter
复制子:	pUC ori,SV40 ori
终止子:	WPRE
质粒分类:	病毒系列质粒;慢病毒类质粒;病毒表达质粒
质粒大小:	8538bp
原核抗性:	Amp（100μg/ml）
筛选标记:	ZsGreen1，Puro
克隆菌株:	Stbl3
培养条件:	37℃，LB，有氧
表达宿主:	293T等哺乳细胞
培养条件:	37℃，5%CO2
诱导方式:	无需诱导
5'测序引物:	CMV-F（CGCAAATGGGCGGTAGGCGTG）
3'测序引物:	EF1-R（CTAGGCACCCGTTCAATTGC）
备注:	本质粒由武汉淼灵生物科技有限公司分享至淼灵质粒平台。

质粒简介

pCDH-ECMV-MCS-3×FLAG-EF1α-ZsGreen1-T2A-Puro质粒是在pCDH载体的基础上改造而成，CMV启动目的基因表达，C端的3×FLAG融合标签能很好的被FLAG抗体识别。该质粒可作为过表达质粒直接转染哺乳细胞，也可包装成慢病毒侵染细胞。质粒本身带有ZsGreen1荧光蛋白和Puro筛选标记，可利用荧光显微镜观察转染效率，也可用嘌呤霉素筛选转染后的细胞得到阳性克隆。

质粒图谱

质粒序列

LOCUS       Exported                8538 bp ds-DNA    circular SYN 10-七月-2017
DEFINITION .
ACCESSION   .
VERSION     .
KEYWORDS    pCDH-ECMV-MCS-3×FLAG-EF1-alpha-ZsGreen1-T2A-Puro
SOURCE      synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE   1 (bases 1 to 8538)
AUTHORS   .
TITLE     Direct Submission
JOURNAL   Exported 2017年7月10日 from MiaoLingPlasmid
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8538
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             234..414
                     /note="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    458..583
                     /note="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1076..1309
                     /note="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     misc_feature    1801..1916
                     /note="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1 (lacking the first T)"
     enhancer        1974..2277
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2278..2481
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     misc_feature    2576..2614
                     /note="MCS"
     CDS             2633..2704
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /note="3xFLAG"
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     promoter        2758..2969
                     /note="EF-1-alpha core promoter"
                     /note="core promoter for human elongation factor
                     EF-1-alpha"
     LTR             2982..3250
                     /note="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             3278..3970
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein"
                     /note="ZsGreen1"
                     /note="mammalian codon-optimized"
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
     CDS             3980..4033
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid
                     protein"
                     /note="T2A"
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /protein_id="
                     "
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             4034..4633
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /note="PuroR"
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     misc_feature    4634..5221
                     /note="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             5295..5528
                     /note="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5600..5721
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
    rep_origin      5740..5875
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     rep_origin      complement(6315..6903)
                     /direction=LEFT
                     /note="ori"
                    /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7074..7934)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7935..8039)
                     /gene="bla"
                     /note="AmpR promoter"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca
     1441 attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg
     1501 aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa
     1561 attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa
     1621 tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt
     1681 ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag
     1741 gtggagagag agacagagac agatccattc gattagtgaa cggatctcga cggtatcgcc
     1801 tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata
     1861 gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg
     1921 gtttattaca gggacagcag agatccagtt tatcgataag cttgggagtt ccgcgttaca
     1981 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     2041 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     2101 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     2161 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     2221 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
     2281 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
     2341 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
     2401 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
     2461 gaggtctata taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca
     2521 cgctgttttg acctccatag aagacaccga ctctactaga ggatctattt ccggtgaatt
     2581 cctcgagact agttctagaa gcggccgcgg atcctcggta ccaagcttaa gtgactacaa
     2641 ggatgacgat gacaaggatt acaaagacga cgatgataag gactataagg atgatgacga
     2701 caaataaaga tccatcgata ctagtaagga tctgcgatcg ctccggtgcc cgtcagtggg
     2761 cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaacgg
     2821 gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc
     2881 tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt
     2941 ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc atctctcctt
     3001 cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc gttctgccgc
     3061 ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta aagctcaggt
     3121 cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc cggctctcca
     3181 cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct gttctgcgcc
     3241 gttacagatc caagctgtga ccggcgccta ctctagcatg gcccagtcca agcacggcct
     3301 gaccaaggag atgaccatga agtaccgcat ggagggctgc gtggacggcc acaagttcgt
     3361 gatcaccggc gagggcatcg gctacccctt caagggcaag caggccatca acctgtgcgt
     3421 ggtggagggc ggccccttgc ccttcgccga ggacatcttg tccgccgcct tcatgtacgg
     3481 caaccgcgtg ttcaccgagt acccccagga catcgtcgac tacttcaaga actcctgccc
     3541 cgccggctac acctgggacc gctccttcct gttcgaggac ggcgccgtgt gcatctgcaa
     3601 cgccgacatc accgtgagcg tggaggagaa ctgcatgtac cacgagtcca agttctacgg
     3661 cgtgaacttc cccgccgacg gccccgtgat gaagaagatg accgacaact gggagccctc
     3721 ctgcgagaag atcatccccg tgcccaagca gggcatcttg aagggcgacg tgagcatgta
     3781 cctgctgctg aaggacggtg gccgcttgcg ctgccagttc gacaccgtgt acaaggccaa
     3841 gtccgtgccc cgcaagatgc ccgactggca cttcatccag cacaagctga cccgcgagga
     3901 ccgcagcgac gccaagaacc agaagtggca cctgaccgag cacgccatcg cctccggctc
     3961 cgccttgccc gcggccgctg agggcagagg aagtcttcta acatgcggtg acgtggagga
     4021 gaatcccggc cctatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
     4081 ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
     4141 cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
     4201 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
     4261 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
     4321 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     4381 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
     4441 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
     4501 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     4561 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     4621 gcccggtgcc tgaaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     4681 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     4741 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     4801 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     4861 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     4921 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     4981 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     5041 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     5101 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     5161 tcttccgcgt ctccgccttc gccctcagac gagtcggatc tccctttggc cgcctccccg
     5221 cctggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
     5281 agaaaagggg ggactggaag ggctaattca ctcccaacga aaataagatc tgctttttgc
     5341 ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     5401 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     5461 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     5521 ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg
     5581 aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa
     5641 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5701 caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc
     5761 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     5821 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac
     5881 ttttgcagag acggcccaaa ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     5941 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     6001 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     6061 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     6121 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     6181 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     6241 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     6301 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6361 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6421 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6481 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     6541 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6601 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6661 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6721 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     6781 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6841 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6901 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6961 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     7021 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     7081 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7141 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7201 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     7261 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     7321 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     7381 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     7441 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7501 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7561 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7621 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7681 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7741 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7801 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     7861 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7921 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7981 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     8041 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     8101 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     8161 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     8221 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     8281 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     8341 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
     8401 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     8461 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     8521 acggccagtg ccaagctg
//

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