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LMBP plasmid details

Last data update: 26 September 2020 04:22 CEST

Plasmid name: pLVX-EF1alpha-Flag-HOIL1-C-Strep-IRES-ZsGreen1 (LMBP 9113)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: -
Sequence
analysis results
Genecorner:

-

Cloned DNA: Human RANBP2-type and C3HC4-type zinc finger containing 1 cDNA (RBCK1, HOIL1, RBCK2, RNF54, UBCE7IP3, XAP4, ZRANB4, GeneID 10616); fragment
FLAG epitope tag; N-terminal
Strep-tag II; C-terminal
Zoanthus sp. green fluorescent protein cDNA (ZsGreen); human-codon-optimised variant (ZsGreen1)
Promoter: Human elongation factor 1α promoter (EF1α)
Phage T7 gene 10 promoter (T7g10)
Phage T3 promoter
Escherichia coli lac operon promoter
Human immunodeficiency virus long terminal repeat (HIV-1 5' LTR)
Ribosome
binding site:
Internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV) polyprotein gene
Terminator: Human immunodeficiency virus long terminal repeat (HIV-1 3' LTR); with deleted U3 region (ΔU3/3' LTR)
Selection marker: Ampicillin (amp)
Replicon: Escherichia coli plasmid pMB1 origin
Host range: Escherichia coli
Mammalian cells
Parental clone: pLVX-EF1alpha-IRES-ZsGreen1; pCDNA3zeo-F-HOIL-C-strep
Further information: The plasmid was constructed by isolating the FLAG- and Strep-tagged N-terminal fragment of the human RBCK1 coding sequence as a PmeI fragment from pCDNA3zeo-F-HOIL-C-strep and cloning it into the pLVX-EF1alpha-IRES-ZsGreen1 vector.
This bicistronic lentiviral expression vector can be used to generate high-titer lentivirus for transducing virtually any dividing or nondividing mammalian cell type, including primary and stem cells. The vector contains an internal ribosomal entry site (IRES) that allows the tagged RBCK1 fragment and the ZsGreen1 fluorescent protein to be simultaneously coexpressed from a single mRNA transcript. Expression of the transcript is driven by the human elongation factor 1 alpha (EF1α) promoter, which continues to be constitutively active even after stable integration of the vector into the host cell genome. Stable expression of the transcript allows the monitoring of a variety of cellular processes (such as differentiation in primary or stem cells) without the transgene silencing associated with CMV promoters. In addition, the vector allows efficient flow cytometric detection of stably or transiently transfected mammalian cells expressing ZsGreen1 and human RBCK1, without time-consuming drug - and clonal selection.
EMBL Accession number: -
Latest sequence update: 26/04/2017
Authenticity test: The plasmid still needs to be subjected to the authenticity test.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: The plasmid was deposited by DR J. Staal(1)(2) and Prof. Dr R. Beyaert(1)(2).
(1) VIB Center for Inflammation Research, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: -
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 DH5α
Host reference: Focus 8 (1986), 9
Related host reference: Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051]
Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187]
Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791]
Rodriguez-Quinones et al., Focus 15 (1993), 110-112
Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660]
Cultivation medium: LB-Lennox + ampicillin (100 μg/ml)
Cultivation temperature: 37°C
Biosafety level: L1 in E. coli; L2 in mammalian cells
Other culture collection numbers: -

Note: Up-to-date, validated data are enclosed with the biological material. Nevertheless, these data are a snapshot at a given moment; further updates are always possible.

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