pLenti-Bi-cistronic

货号 名称 规格 小包装 大包装
P0270 pLenti-Bi-cistronic 质粒干粉 QQ询价 QQ询价
亲爱的科研工作者,感谢您信任淼灵质粒平台,我们致力于为大家分享质优价适的质粒,我们的各批次质粒干粉在发货前均经过严格的多重验证,您收到后请先转化,再挑取单克隆扩增,万一存在质量问题,请及时通知我们退换。如本页面质粒信息不全,请及时联系所在区域负责人QQ加急上传。
启动子: RSV/PGK
复制子: M13
终止子: SV40 poly(A) signal
质粒分类: 病毒系列,慢病毒双表达载体
质粒大小: 8292bp
原核抗性: Kan
真核抗性: Puro
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,瞬时表达
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
片段类型: 基因CDS
片段物种: 双框空载体
原核抗性: 卡那霉素
真核抗性: 嘌呤霉素
荧光标记:
    This study was aimed to construct, package and purify the recombinant lentivirus vector carrying the firefly luciferase gene (FLUC) and red fluorescent protein gene (RFP) and to transfect the recombinant lentivirus into HeLa cells, so as to observe the expression levels of these two genes. The FLUC and RFP genes were amplified by RT-PCR and inserted in the lentiviral expression vector (pLenti-Bi-cistronic) to construct the lentiviral vector pLenti-FLUC-RFP. The viral particles were generated by cotransfection of 293T cells with pLenti-FLUC-RFP and three packaging vectors, and the virus titer was determined by calculating the percentage of RFP positive cells. After transfection of pLenti-FLUC-RFP into HeLa cells, the expression of RFP was observed by fluorescent microscopy, and the activity of FLUC was determined by luciferase reporter gene assay kit. The results showed that the inserting orientation of the RFP and FLUC genes in the lentiviral vector pLenti-FLUC-RFP were verified by restriction analysis. Targeted RFP and FLUC sequences were confirmed by DNA sequencing. The final titer obtained was 1×10TU/ml. The expressions of RFP and FLUC were observed in the transfected HeLa cells. It is concluded that the pLenti-III-FLUC-RFP recombinant lentivirus vector carrying RFP gene and FLUC gene with high viral titer is constructed and packaged successfully, and provides experimental basis for studying dynamic distribution of mesenchymal stem cells in vivo.
LOCUS       Exported                8292 bp ds-DNA     circular SYN 08-SEP-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Untitled 2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8292)
  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..8292
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_signal     170..320
                     /note="Ad5 Psi"
                     /note="packaging signal for adenovirus serotype 5"
     promoter        488..714
                     /note="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             715..895
                     /note="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    942..1067
                     /note="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1557..1790
                     /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    2297..2412
                     /note="cPPT/CTS"
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     polyA_signal    2441..2522
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     promoter        2733..3232
                     /note="PGK promoter"
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             3309..3335
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /note="HA"
                     /translation="YPYDVPDYA"
     promoter        3353..3650
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      3520..3655
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     CDS             3656..4255
                     /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    4268..4855
                     /note="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(4739..4750)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /note="Factor Xa site"
                     /translation="IEGR"
     LTR             4865..5098
                     /note="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5170..5291
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(5619..6207)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             7014..7808
                     /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(7856..8235)
                     /direction=LEFT
                     /note="M13 ori"
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     repeat_region   8273..82
                     /note="ITR"
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
ORIGIN
        1 ttttggattg aagccaatat gataatgagg gggtggagtt tgtgacgtgg cgcggggcgt
       61 gggaacgggg cgggtgacgt agtagtgtgg cggaagtgtg atgttgcaag tgtggcggaa
      121 cacatgtaag cgacggatgt ggcaaaagtg acgtttttgg tgtgcgccgg tgtacacagg
      181 aagtgacaat tttcgcgcgg ttttaggcgg atgttgtagt aaatttgggc gtaaccgagt
      241 aagatttggc cattttcgcg ggaaaactga ataagaggaa gtgaaatctg aataattttg
      301 tgttactcat agcgcgtaat acggcagacc tcagcgctag attattgaag catttatcag
      361 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
      421 gttccgcgca catttccccg aaaagtgcca cctgacgtta actataacgg tcctaaggta
      481 gcgaaaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag
      541 caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg
      601 tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact
      661 gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct
      721 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      781 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      841 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
      901 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
      961 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1021 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1081 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1141 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1201 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1261 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1321 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1381 aaagtaagac caccgcacag caagcccgct gatcttcaga cctggaggag gagatatgag
     1441 ggacattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     1501 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     1561 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     1621 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     1681 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     1741 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     1801 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     1861 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     1921 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     1981 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     2041 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     2101 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     2161 tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa
     2221 ggtggagaga gagacagaga cagatccatt cgattagtga acggatctcg acggtatcga
     2281 aagcttggga ttcgaattta aaagaaaagg ggggattggg gggtacagtg caggggaaag
     2341 aatagtagac ataatagcaa cagacataca aactaaagaa ctacaaaaac aaattacaaa
     2401 aattcaaaat tttcgggttt ttcgaaccta gcagacgtcg gcagtgaaaa aaatgcttta
     2461 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     2521 ttaacaacaa gaattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     2581 tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga ttatgatctc acctaggctc
     2641 gagggatccc ccgggcgatc tgacggttca ctaaacgagc tctgcttata taggcctccc
     2701 accgtacacg ccacctcgac ataaattcta ccgggtaggg gaggcgcttt tcccaaggca
     2761 gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg
     2821 gcctcgcaca cattccacat ccaccggtag gcgccaaccg gctccgttct ttggtggccc
     2881 cttcgcgcca ccttctactc ctcccctagt caggaagttc ccccccgccc cgcagctcgc
     2941 gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact agtctcgtgc agatggacag
     3001 caccgctgag caatggaagc gggtaggcct ttggggcagc ggccaatagc agctttgctc
     3061 cttcgctttc tgggctcaga ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg
     3121 gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc ccggcattct gcacgcttca
     3181 aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct ccgggccttt cgacctatcg
     3241 atcaattgag tacttacgta ggtaccgaat tcgatatcgg gcccgcggcc gcgtctagat
     3301 gagtcgagta cccatacgac gtcccagact acgcttgagt ttaaacacgc gtggtgtgga
     3361 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3421 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3481 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3541 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3601 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggccatgac
     3661 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccctcggg ccgtacgcac
     3721 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtggacc cggaccgcca
     3781 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     3841 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt
     3901 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     3961 gctggccgcg cagcaacaga tggaagggct cctggcgccg caccggccca aggagcccgc
     4021 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc
     4081 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     4141 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     4201 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaacgcg
     4261 ttccggaaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     4321 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     4381 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     4441 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     4501 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     4561 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     4621 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     4681 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     4741 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     4801 gcgtctcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgtc
     4861 cggatggaag ggctaattca ctcccaacga atacaagatc tgctttttgc ttgtactggg
     4921 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     4981 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     5041 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
     5101 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag
     5161 agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     5221 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     5281 aatgtatctt atcatgtctg gcatctatgt cgggtgcgga gaaagaggta atgaaatggc
     5341 attatgggta ttatgggtct gcattaatga atcggccaac gatcccggtg tgaaataccg
     5401 cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac
     5461 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5521 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5581 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5641 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5701 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5761 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5821 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5881 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5941 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6001 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     6061 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6121 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6181 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6241 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6301 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6361 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6421 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6481 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6541 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6601 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6661 gttaatagtt tgcgcaacgt tgttgaaaaa ggatcttcac ctagatcctt ttcacgtaga
     6721 aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt cagctactgg gctatctgga
     6781 caagggaaaa cgcaagcgca aagagaaagc aggtagcttg cagtgggctt acatggcgat
     6841 agctagactg ggcggtttta tggacagcaa gcgaaccgga attgccagct ggggcgccct
     6901 ctggtaaggt tgggaagccc tgcaaagtaa actggatggc tttctcgccg ccaaggatct
     6961 gatggcgcag gggatcaagc tctgatcaag agacaggatg aggatcgttt cgcatgattg
     7021 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     7081 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     7141 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     7201 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     7261 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     7321 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     7381 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     7441 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     7501 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     7561 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     7621 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     7681 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     7741 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     7801 tcttctgaat tttgttaaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     7861 atcggcaaca tcccttataa atcaaaagaa tagaccgcga tagggttgag tgttgttcca
     7921 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     7981 gtctatcagg gcgatggccc actacgtgaa ccatcaccca aatcaagttt tttgcggtcg
     8041 aggtgccgta aagctctaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     8101 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     8161 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgcgcg cttaatgcgc
     8221 cgctacaggg cgcgtccatt cgccattcag gatcgaatta attcttaatt aacatcatca
     8281 ataatatacc tt
//