pCDH-EF1-MCS-T2A-copGFP

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别名: pCDH-EF1a-MCS-T2A-copGFP
启动子: EF1α
复制子: pUC
终止子: SV40 poly(A) signal
质粒分类: 病毒系列,慢病毒克隆载体
质粒大小: 7266bp
原核抗性: Amp
克隆菌株: Stbl3
培养条件: 37℃
表达宿主: 哺乳细胞
诱导方式: 无须诱导
5'测序引物: 根据序列设计引物
3'测序引物: 根据序列设计引物
质粒宿主: 哺乳细胞,慢病毒
质粒用途: 蛋白表达
质粒类型: 基因CDS
基因物种:
原核抗性: 氨苄青霉素
筛选标记:
荧光标记: 绿色
    Stable overexpression with cDNA lentivectors Strong and ubiquitous expression of the gene of interestSingle or double expression cassette with choice of reporter geneTarget gene expressed from CMV, EF1, or MSCV promoter Choose from FIV- or HIV-based vectors Gene expression from constitutive promoters,The multiple cloning site (MCS) located downstream of an internal promoter allows convenient cloning of your gene of interest.
LOCUS       Exported                7266 bp ds-DNA     circular SYN 05-AUG-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pCDH-EF1-MCS-T2A-copGFP (CD521A-1)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7266)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, September 8, 2016 from SnapGene Viewer 3.1.4
            http://www.miaolingbio.com
FEATURES             Location/Qualifiers
     source          1..7266
                     /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    1804..1921
                     /note="cPPT/CTS"
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1966..2177
                     /note="EF-1-alpha core promoter"
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             2190..2458
                     /note="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             2524..2577
                     /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."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             2608..3270
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina 
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /note="CopGFP"
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
     misc_feature    3361..3949
                     /note="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(3832..3843)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /note="Factor Xa site"
                     /translation="IEGR"
     LTR             4023..4256
                     /note="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4328..4449
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      4468..4603
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     primer_bind     complement(4641..4657)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4665..4681
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /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)."
     promoter        complement(4689..4719)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    4734..4755
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5043..5631)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5802..6662)
                     /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(6663..6767)
                     /gene="bla"
                     /note="AmpR promoter"
     primer_bind     7241..7257
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
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 gggcgcagcc 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 acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     1921 tatcgatact agtaaggatc tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat
     1981 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaacgggt gcctagagaa
     2041 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
     2101 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
     2161 ttgccgccag aacacagctg aagcttcgag gggctcgcat ctctccttca cgcgcccgcc
     2221 gccctacctg aggccgccat ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg
     2281 gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg agaccgggcc
     2341 tttgtccggc gctcccttgg agcctaccta gactcagccg gctctccacg ctttgcctga
     2401 ccctgcttgc tcaactctac gtctttgttt cgttttctgt tctgcgccgt tacagatcca
     2461 agctgtgacc ggcgcctact ctagagctag cgaattcgaa tttaaatcgg atccgcggcc
     2521 gctgagggca gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc cggcccttcc
     2581 ggaatggaga gcgacgagag cggcctgccc gccatggaga tcgagtgccg catcaccggc
     2641 accctgaacg gcgtggagtt cgagctggtg ggcggcggag agggcacccc caagcagggc
     2701 cgcatgacca acaagatgaa gagcaccaaa ggcgccctga ccttcagccc ctacctgctg
     2761 agccacgtga tgggctacgg cttctaccac ttcggcacct accccagcgg ctacgagaac
     2821 cccttcctgc acgccatcaa caacggcggc tacaccaaca cccgcatcga gaagtacgag
     2881 gacggcggcg tgctgcacgt gagcttcagc taccgctacg aggccggccg cgtgatcggc
     2941 gacttcaagg tggtgggcac cggcttcccc gaggacagcg tgatcttcac cgacaagatc
     3001 atccgcagca acgccaccgt ggagcacctg caccccatgg gcgataacgt gctggtgggc
     3061 agcttcgccc gcaccttcag cctgcgcgac ggcggctact acagcttcgt ggtggacagc
     3121 cacatgcact tcaagagcgc catccacccc agcatcctgc agaacggggg ccccatgttc
     3181 gccttccgcc gcgtggagga gctgcacagc aacaccgagc tgggcatcgt ggagtaccag
     3241 cacgccttca agacccccat cgccttcgcc agatcccgcg ctcagtcgtc caattctgcc
     3301 gtggacggca ccgccggacc cggctccacc ggatctcgct agagctgaat ctaagtcgac
     3361 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     3421 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     3481 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     3541 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     3601 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     3661 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     3721 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     3781 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     3841 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     3901 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggtaccttt
     3961 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg
     4021 actggaaggg ctaattcact cccaacgaaa ataagatctg ctttttgctt gtactgggtc
     4081 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     4141 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     4201 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag
     4261 tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag
     4321 tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4381 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4441 tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccag ttccgcccat
     4501 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     4561 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagactt ttgcagagac
     4621 ggcccaaatt cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     4681 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     4741 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4801 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4861 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4921 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4981 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5041 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5101 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5161 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5221 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5281 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5341 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5401 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5461 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     5521 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5581 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5641 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5701 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5761 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5821 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5881 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5941 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6001 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6061 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6121 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6181 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6241 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6301 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6361 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6421 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6481 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6541 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     6601 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6661 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6721 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6781 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     6841 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     6901 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     6961 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     7021 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
     7081 agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     7141 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     7201 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc
     7261 aagctg
//