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pLVX-EF1伪-IRES-mCherry/ pLVX-EF1伪-IRES-mCherry质粒/ pLVX-EF1伪-IRES-mCherry Plasmid

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  • pLVX-EF1α-IRES-mCherry

    PVT2309    2ug

     

    pLVX-EF1α-IRES-mCherry Informaiton

    Promoter: EF-1 alpha promoter

    Replicon: pUC ori

    Terminator: SV40 poly (A) signal

    Plasmid classification: virus series, lentivirus cloning vector

    Prokaryotic resistance: ampicillin Amp

    Selection marker: red fluorescence

    Clone strain: Escherichia coli Stbl3

    Culture conditions: 37 C, aerobic, LB

    Expression host: lactation cells

    Induction mode: no need to induce, transient expression.

    5'sequencing primers: WPRE-R:CATAGCGTAAAAGGAGCAACA

    Primers for 3'sequencing: primers designed based on sequences

    Remarks: there is a difference from the sequence of Clontech.

     

     

    pLVX-EF1α-IRES-mCherry Description

    Plvx-ef1 alpha-IRES mCherry is a bigenic lentiviral expression vector that can be used to transduce mammalian cells to produce high titers of lentiviruses, including primary and stem cells, in almost any division or non-division. The vector contains an internal ribosome entry site (IRES) that allows an interesting gene and red fluorescent protein mCherry to be simultaneously transcribed from a single mRNA expression. Transcriptionally expressed human elongation factor-1 alpha (EF-1 alpha) promoters are driven and continue to be activated even when the genome of the host cell is stably integrated into the vector. Stable transcription allows the monitoring of various cellular processes, such as primary or stem cell differentiation, not to be silenced with CMV promoters. In addition, vectors allow efficient flow cytometry to detect the expressed proteins, mCherry, and benefits of stable or transient transfection of mammalian cells without time-consuming drug and cloning options.

     

    pLVX-EF1α-IRES-mCherry Sequence

    LOCUS       Exported                8935 bp ds-DNA     circular SYN 07-SEP-2016
    DEFINITION  synthetic circular DNA.
    ACCESSION   .
    VERSION     .
    KEYWORDS    pLVX-EF1-alpha-IRES-mCherry
    SOURCE      synthetic DNA construct
      ORGANISM  synthetic DNA construct
    REFERENCE   1  (bases 1 to 8935)
      AUTHORS   .
      TITLE     Direct Submission
      JOURNAL   Exported Wednesday, September 7, 2016 from SnapGene Viewer 3.1.4
         
    FEATURES             Location/Qualifiers
         source          1..8935
                         /organism="synthetic DNA construct"
                         /mol_type="other DNA"
         LTR             1..634
                         /note="3' LTR"
                         /note="3' long terminal repeat (LTR) from HIV-1"
         misc_feature    681..806
                         /note="HIV-1 Psi"
                         /note="packaging signal of human immunodeficiency virus 
                         type 1"
         misc_feature    1303..1536
                         /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    2028..2143
                         /note="cPPT/CTS"
                         /note="central polypurine tract and central termination 
                         sequence of HIV-1 (lacking the first T)"
         promoter        2338..3519
                         /note="EF-1-alpha promoter"
                         /note="strong constitutive promoter for human elongation 
                         factor EF-1-alpha"
         intron          2568..3510
                         /note="EF-1-alpha intron A"
                         /note="intron upstream of the start codon of human 
                         EF-1-alpha"
         misc_feature    3575..4147
                         /note="IRES"
                         /note="internal ribosome entry site (IRES) of the 
                         encephalomyocarditis virus (EMCV)"
         CDS             4180..4890
                         /codon_start=1
                         /product="monomeric derivative of DsRed fluorescent protein
                         (Shaner et al., 2004)"
                         /note="mCherry"
                         /note="mammalian codon-optimized"
                         /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                         TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                         EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                         GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                         EGRHSTGGMDELYK"
         misc_feature    4904..5492
                         /note="WPRE"
                         /note="woodchuck hepatitis virus posttranscriptional 
                         regulatory element"
         CDS             complement(5375..5386)
                         /codon_start=1
                         /product="Factor Xa recognition and cleavage site"
                         /note="Factor Xa site"
                         /translation="IEGR"
         LTR             5699..6332
                         /note="3' LTR"
                         /note="3' long terminal repeat (LTR) from HIV-1"
         primer_bind     complement(6460..6476)
                         /note="M13 rev"
                         /note="common sequencing primer, one of multiple similar 
                         variants"
         protein_bind    6484..6500
                         /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(6508..6538)
                         /note="lac promoter"
                         /note="promoter for the E. coli lac operon"
         protein_bind    6553..6574
                         /bound_moiety="E. coli catabolite activator protein"
                         /note="CAP binding site"
                         /note="CAP binding activates transcription in the presence 
                         of cAMP."
         rep_origin      complement(6862..7450)
                         /direction=LEFT
                         /note="ori"
                         /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                         replication"
         CDS             complement(7621..8481)
                         /codon_start=1
                         /gene="bla"
                         /product="beta-lactamase"
                         /note="AmpR"
                         /note="confers resistance to ampicillin, carbenicillin, and
                         related antibiotics"
                         /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                         ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                         PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
    温馨提示:不可用于临床ZL。
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