pLVshRNA-EGFP(2A)Puro

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P0684 pLVshRNA-EGFP(2A)Puro 质粒干粉 请加QQ咨询 请加QQ咨询
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启动子: U6
复制子: pUC ori
终止子: SV40 poly(A) signal
原核抗性: 氨苄青霉素Amp
筛选标记: 嘌呤霉素Puro
克隆菌株: 大肠杆菌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: U6:ATGGACTATCATATGCTTACCGTA
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:
         pLVshRNA-EGFP(2A)Puro质粒是基于HIV1的慢病毒RNA干扰载体,用于表达人工设计合成的shRNA,干扰目标基因的表达。
LOCUS       Exported                8593 bp ds-DNA     circular SYN 28-DEC-2017
DEFINITION  synthetic circular DNA
FEATURES             Location/Qualifiers
     source          1..8593
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    2028..2143
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     promoter        2196..2436
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     promoter        2510..3009
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             3036..3752
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             3768..3827
                     /codon_start=1
                     /product="2A peptide from equine rhinitis A virus 
                     polyprotein"
                     /label=E2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="QCTNYALLKLAGDVESNPGP"
     CDS             3828..4427
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     misc_feature    4521..5106
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             5310..5943
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6473..7061)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7232..8092)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8093..8197)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    8245..8379
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgatctgggc aggaagaggg cctatttccc atgattcctt
     2221 catatttgca tatacgatac aaggctgtta gagagataat tagaattaat ttgactgtaa
     2281 acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg
     2341 cagttttaaa attatgtttt aaaatggact atcatatgct taccgtaact tgaaagtatt
     2401 tcgatttctt ggctttatat atcttgtgga aaggacgagg atccgcctaa ggttaagtcg
     2461 ccctcgctcg agcgagggcg acttaacctt aggttttttg aattctaccg ggtaggggag
     2521 gcgcttttcc caaggcagtc tggagcatgc gctttagcag ccccgctggg cacttggcgc
     2581 tacacaagtg gcctctggcc tcgcacacat tccacatcca ccggtaggcg ccaaccggct
     2641 ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag gaagttcccc
     2701 cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg tctcactagt
     2761 ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc
     2821 caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg
     2881 ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg
     2941 gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg
     3001 ggcctttcga cctgcagccc aagcttaccg ccaccatggt gagcaagggc gaggagctgt
     3061 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     3121 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     3181 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
     3241 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     3301 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     3361 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     3421 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     3481 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     3541 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     3601 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
     3661 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     3721 ggatcactct cggcatggac gagctgtaca aggaatttgg cagcggccag tgcaccaact
     3781 atgccctgct gaaactggct ggcgatgtgg aaagcaaccc cggacccatg accgagtaca
     3841 agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg
     3901 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc
     3961 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
     4021 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
     4081 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
     4141 cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
     4201 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
     4261 tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
     4321 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
     4381 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgacgc ccgccccacg
     4441 acccgcagcg cccgaccgaa aggagcgcac gaccccatgc atcggcgtct cgagtctaga
     4501 gggcccgacg cgtctggaac aatcaacctc tggattacaa aatttgtgaa agattgactg
     4561 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atggctttgt
     4621 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     4681 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     4741 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     4801 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     4861 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga
     4921 cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     4981 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     5041 tgcggcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     5101 ccccgcctgg aattaattct gcagtcgaga cctagaaaaa catggagcaa tcacaagtag
     5161 caatacagca gctatcaatg attgtgcctg gctagaagca caagaggagg aggaggtggg
     5221 ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct
     5281 tagccacttt ttaaaagaaa agaggggact ggaagggcta attcactccc aacgaagaca
     5341 agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt agcagaacta
     5401 cacaccaggg ccaggggtca gatatccact gacctttgga tggtgctaca agctagtacc
     5461 agttgagcca gataaggtag aagaggccaa taaaggagag aacaccagct tgttacaccc
     5521 tgtgagcctg catgggatgg atgacccgga gagagaagtg ttagagtgga ggtttgacag
     5581 ccgcctagca tttcatcacg tggcccgaga gctgcatccg gagtacttca agaactgctg
     5641 atatcgagct tgctacaagg gactttccgc tggggacttt ccagggaggc gtggcctggg
     5701 cgggactggg gagtggcgag ccctcagatc ctgcatataa gcagctgctt tttgcctgta
     5761 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     5821 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     5881 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     5941 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat
     6001 cagagagtga gaggccttga cattgctagc gtttaccgtc gacctctagc tagagcttgg
     6061 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     6121 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     6181 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     6241 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     6301 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     6361 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     6421 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6481 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6541 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6601 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6661 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6721 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6781 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6841 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6901 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6961 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     7021 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     7081 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     7141 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     7201 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     7261 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     7321 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     7381 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     7441 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     7501 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     7561 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     7621 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     7681 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7741 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     7801 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     7861 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7921 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7981 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     8041 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     8101 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     8161 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     8221 ctgacgtcga cggatcggga gatcaacttg tttattgcag cttataatgg ttacaaataa
     8281 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     8341 ttgtccaaac tcatcaatgt atcttatcat gtctggatca actggataac tcaagctaac
     8401 caaaatcatc ccaaacttcc caccccatac cctattacca ctgccaatta cctgtggttt
     8461 catttactct aaacctgtga ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt
     8521 taaatatgta ctacaaactt agtagttttt aaagaaattg tatttgttaa atatgtacta
     8581 caaacttagt agt
//