pOTENT-CMV-Flag-His-Puro

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启动子: CMV,T7
复制子: M13 ori,ori ,SV40 ori
终止子: bGH poly(A) signal
质粒分类: 腺病毒表达质粒
质粒大小: 7509bp
原核抗性: Kan
筛选标记: Neo
克隆菌株: Stbl3
培养条件: 37℃,有氧 LB
表达宿主: 哺乳细胞
诱导方式: 无须诱导,瞬时表达
5'测序引物: CMV-F:CGCAAATGGGCGGTAGGCGTG
3'测序引物: 根据序列设计引物
质粒宿主:
质粒用途:
质粒类型:
基因物种:
原核抗性:
筛选标记:
荧光标记:
pOTENT-CMV-Flag-His-Puro
LOCUS       Exported                7509 bp ds-DNA     circular SYN 12-JAN-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pOTENT-CMV-Flag-His-Puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7509)
  AUTHORS   Huanghua
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, September 8, 2016 from SnapGene Viewer 3.1.4
            http://www.miaolingbio.com
FEATURES             Location/Qualifiers
     source          1..7509
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   21..123
                     /note="ITR"
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     misc_signal     211..361
                     /note="Ad5 Psi"
                     /note="packaging signal for adenovirus serotype 5"
     enhancer        419..722
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        723..926
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        954..972
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1073..1096
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /note="FLAG"
                     /translation="DYKDDDDK"
     CDS             1106..1123
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     polyA_signal    1294..1415
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     promoter        2342..2671
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      2522..2657
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     CDS             2729..3328
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /note="PuroR"
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    3329..3553
                     /note="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4464..4566
                     /note="ITR"
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     rep_origin      complement(4838..5426)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             6258..7052
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="NeoR/KanR"
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     rep_origin      complement(7100..7480)
                     /direction=LEFT
                     /note="M13 ori"
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 ttcgaattta aattaattaa catcatcaat aatatacctt attttggatt gaagccaata
       61 tgataatgag ggggtggagt ttgtgacgtg gcgcggggcg tgggaacggg gcgggtgacg
      121 tagtagtgtg gcggaagtgt gatgttgcaa gtgtggcgga acacatgtaa gcgacggatg
      181 tggcaaaagt gacgtttttg gtgtgcgccg gtgtacacag gaagtgacaa ttttcgcgcg
      241 gttttaggcg gatgttgtag taaatttggg cgtaaccgag taagatttgg ccattttcgc
      301 gggaaaactg aataagagga agtgaaatct gaataatttt gtgttactca tagcgcgtaa
      361 tactgtaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      601 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      661 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      721 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      841 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      901 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagtaatacg
      961 actcactata gggagaggat ccggtaccga ggagatctgc cgccgcgatc gccggcgcgc
     1021 cagatctcaa gcttaactag ctagcggacc gacgcgtacg cggccgctcg aggattataa
     1081 ggatgacgac gataaattcg tcgagcacca ccaccaccac cactaataag gtttatccga
     1141 tccaccggat ctagataaga tatccgatcc accggatcta gataactgat cataatcagc
     1201 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     1261 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     1321 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     1381 agttgtggtt tgtccaaact catcaatgta tcttaacgcg gatctgggcg tggttaaggg
     1441 tgggaaagaa tatataaggt gggggtctta tgtagttttg tatctgtttt gcagcagccg
     1501 ccgccgccat gagcaccaac tcgtttgatg gaagcattgt gagctcatat ttgacaacgc
     1561 gcatgccccc atgggccggg gtgcgtcaga atgtgatggg ctccagcatt gatggtcgcc
     1621 ccgtcctgcc cgcaaactct actaccttga cctacgagac cgtgtctgga acgccgttgg
     1681 agactgcagc ctccgccgcc gcttcagccg ctgcagccac cgcccgcggg attgtgactg
     1741 actttgcttt cctgagcccg cttgcaagca gtgcagcttc ccgttcatcc gcccgcgatg
     1801 acaagttgac ggctcttttg gcacaattgg attctttgac ccgggaactt aatgtcgttt
     1861 ctcagcagct gttggatctg cgccagcagg tttctgccct gaaggcttcc tcccctccca
     1921 atgcggttta aaacataaat aaaaaaccag actctgtttg gatttggatc aagcaagtgt
     1981 cttgctgtct ttatttaggg gttttgcgcg cgcggtaggc ccgggaccag cggtctcggt
     2041 cgttgagggt cctgtgtatt ttttccagga cgtggtaaag gtgactctgg atgttcagat
     2101 acatgggcat aagcccgtct ctggggtgga ggtagcacca ctgcagagct tcatgctgcg
     2161 gggtggtgtt gtagatgatc cagtcgtagc aggagcgctg ggcgtggtgc ctaaaaatgt
     2221 ctttcagtag caagctgatt gccaggggca ggcccttggt gtaagtgttt acaaagcggt
     2281 taagctggga tgggtgcata cgtgtttaaa ctagggataa cagggtaatc ggctgtggaa
     2341 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2401 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     2461 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     2521 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     2581 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     2641 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     2701 cggatctgat caagagacac gtacgaccat gaccgagtac aagcccacgg tgcgcctcgc
     2761 cacccgcgac gacgtccccc gggcagtacg caccctcgcc gccgcgttcg ccgactaccc
     2821 cgccacgcgc cacaccgtcg atccagaccg ccacatcgag cgggtcaccg agctgcaaga
     2881 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     2941 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     3001 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     3061 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     3121 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     3181 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     3241 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     3301 gtgcatgacc cgcaagcccg gtgcctgact gtgccttcta gttgccagcc atctgttgtt
     3361 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     3421 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     3481 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     3541 gtgggctcta tggtagggat aacagggtaa tgtttaaact ataaaatgca aggtgctgct
     3601 caaaaaatca ggcaaagcct cgcgcaaaaa agaaagcaca tcgtagtcat gctcatgcag
     3661 ataaaggcag gtaagctccg gaaccaccac agaaaaagac accatttttc tctcaaacat
     3721 gtctgcgggt ttctgcataa acacaaaata aaataacaaa aaaacattta aacattagaa
     3781 gcctgtctta caacaggaaa aacaaccctt ataagcataa gacggactac ggccatgccg
     3841 gcgtgaccgt aaaaaaactg gtcaccgtga ttaaaaagca ccaccgacag ctcctcggtc
     3901 atgtccggag tcataatgta agactcggta aacacatcag gttgattcac atcggtcagt
     3961 gctaaaaagc gaccgaaata gcccggggga atacataccc gcaggcgtag agacaacatt
     4021 acagccccca taggaggtat aacaaaatta ataggagaga aaaacacata aacacctgaa
     4081 aaaccctcct gcctaggcaa aatagcaccc tcccgctcca gaacaacata cagcgcttcc
     4141 acagcggcag ccataacagt cagccttacc agtaaaaaag aaaacctatt aaaaaaacac
     4201 cactcgacac ggcaccagct caatcagtca cagtgtaaaa aagggccaag tgcagagcga
     4261 gtatatatag gactaaaaaa tgacgtaacg gttaaagtcc acaaaaaaca cccagaaaac
     4321 cgcacgcgaa cctacgccca gaaacgaaag ccaaaaaacc cacaacttcc tcaaatcgtc
     4381 acttccgttt tcccacgtta cgtcacttcc cattttaaga aaactacaat tcccaacaca
     4441 tacaagttac tccgccctaa aacctacgtc acccgccccg ttcccacgcc ccgcgccacg
     4501 tcacaaactc caccccctca ttatcatatt ggcttcaatc caaaataagg tatattattg
     4561 atgatgttaa ttaatttaaa ttcgaacatg catggatcca tatgcggtgt gaaataccgc
     4621 acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact
     4681 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4741 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4801 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4861 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4921 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4981 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5041 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5101 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5161 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5221 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     5281 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5341 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5401 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5461 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5521 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5581 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5641 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5701 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5761 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5821 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5881 ttaatagttt gcgcaacgtt gttgccattg ctgcagccat gagattatca aaaaggatct
     5941 tcacctagat ccttttcacg tagaaagcca gtccgcagaa acggtgctga ccccggatga
     6001 atgtcagcta ctgggctatc tggacaaggg aaaacgcaag cgcaaagaga aagcaggtag
     6061 cttgcagtgg gcttacatgg cgatagctag actgggcggt tttatggaca gcaagcgaac
     6121 cggaattgcc agctggggcg ccctctggta aggttgggaa gccctgcaaa gtaaactgga
     6181 tggctttctt gccgccaagg atctgatggc gcaggggatc aagctctgat caagagacag
     6241 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     6301 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     6361 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     6421 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     6481 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     6541 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     6601 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     6661 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     6721 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     6781 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     6841 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     6901 ccgaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     6961 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     7021 ccttctatcg ccttcttgac gagttcttct gaattttgtt aaaatttttg ttaaatcagc
     7081 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     7141 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     7201 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     7261 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     7321 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     7381 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     7441 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt ccattcgcca ttcaggatcg
     7501 aattaattc
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