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pLVX- IRES- Neomycin/ pLVX- IRES- Neomycin质粒/ pLVX- IRES- Neomycin Plasmid

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  • pLVX-IRES-Neomycin

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    pLVX-IRES-Neomycin,Plasmid pLVX-IRES-Neomycin,pLVX-IRES-Neomycin vector

    pLVX-IRES-Neomycin Plasmid information

    Promoter: CMV

    Replicon: pUC ori

    Terminator: SV40 poly (A) signal

    Plasmid classification: virus series, lentivirus cloning vector

    Plasmid size: 8269bp

    Prokaryotic resistance: Amp

    Screening markers: Neo

    Cloned strain: Stbl3

    Culture conditions: 37 centigrade, aerobic LB

    Expression host: mammalian cells

    Induction mode: no induction, instantaneous expression

    5'sequencing primers: CMV-F:CGCAAATGGGCGGTAGGCGTG

    3'sequencing primers: primers designed according to sequence

    Use:Lentiviral

    pLVX-IRES-Neomycin Plasmid Description

    pLVX-IRES-Neo is an HIV-1-based, lentiviral expression vector that allows the simultaneous expression of your protein of interest and G418 resistance (Neor) in virtually any mammalian cell type, including primary cells. The vector expresses your protein of interest and Neor from a bicistronic mRNA transcript, allowing G418 resistance to be used as an indicator of transduction efficiency and a marker for selection. pLVX-IRES-Neo contains all of the viral processing elements necessary for the production of replication-incompetent lentivirus, as well as elements to improve viral titer, transgene expression, and overall vector function. The woodchuck hepatitis virus posttranscriptional regulatory element (WPRE) promotes RNA processing events and enhances nuclear export of viral RNA , leading to increased viral titers from packaging cells. In addition, the vector includes a Rev-response element (RRE), which further increases viral titers by enhancing the transport of unspliced viral RNA out of the nucleus . Finally, pLVX-IRES-Neo also contains a central polypurine tract/central termination sequence element (cPPT/CTS). During target cell infection, this element creates a central DNA flap that increases nuclear import of the viral genome, resulting in improved vector integration and more efficient transduction . The vector also contains a pUC origin of replication and an E. coli ampicillin resistance gene (Ampr) for propagation and selection in bacteria

    pLVX-IRES-Neomycin Plasmid Sequence
     

    LOCUS       Exported                8269 bp ds-DNA     circular SYN 27-AUG-2016
    DEFINITION  synthetic circular DNA
    ACCESSION   .
    VERSION     .
    KEYWORDS    Untitled 20
    SOURCE      synthetic DNA construct
      ORGANISM  synthetic DNA construct
    REFERENCE   1  (bases 1 to 8269)
      AUTHORS   .
      TITLE     Direct Submission
      JOURNAL   Exported Saturday, August 27, 2016 from SnapGene Viewer 3.1.4
                
    FEATURES             Location/Qualifiers
         source          1..8269
                         /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)"
         enhancer        2201..2504
                         /note="CMV enhancer"
                         /note="human cytomegalovirus immediate early enhancer"
         promoter        2505..2708
                         /note="CMV promoter"
                         /note="human cytomegalovirus (CMV) immediate early 
                         promoter"
         misc_feature    2843..3416
                         /note="IRES"
                         /note="internal ribosome entry site (IRES) of the 
                         encephalomyocarditis virus (EMCV)"
         CDS             3430..4224
                         /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="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                         VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                         SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                         GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                         LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
         misc_feature    4238..4826
                         /note="WPRE"
                         /note="woodchuck hepatitis virus posttranscriptional 
                         regulatory element"
         CDS             complement(4709..4720)
                         /codon_start=1
                         /product="Factor Xa recognition and cleavage site"
                         /note="Factor Xa site"
                         /translation="IEGR"
         LTR             5033..5666
                         /note="3' LTR"
                         /note="3' long terminal repeat (LTR) from HIV-1"
         primer_bind     complement(5794..5810)
                         /note="M13 rev"
                         /note="common sequencing primer, one of multiple similar 
                         variants"
         protein_bind    5818..5834
                         /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(5842..5872)
                         /note="lac promoter"
                         /note="promoter for the E. coli lac operon"
         protein_bind    5887..5908
                         /bound_moiety="E. coli catabolite activator protein"
                         /note="CAP binding site"
                         /note="CAP binding activates transcription in the presence 
                         of cAMP."
         rep_origin      complement(6196..6784)
                         /direction=LEFT
                         /note="ori"
                         /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                         replication"
         CDS             complement(6955..7815)
                         /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(7816..7920)
                         /gene="bla"
                         /note="AmpR promoter"
         polyA_signal    7968..8102
                         /note="SV40 poly(A) signal"
                         /note="SV40 polyadenylation signal"
    ORIGIN
            1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
           61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
    温馨提示:不可用于临床ZL。
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