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LMBP plasmid details

Last data update: 26 September 2020 04:22 CEST

Plasmid name: pLVX-EF1alpha-hCARMA2-H171N-IRES-mCherry (LMBP 9628)

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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 caspase recruitment domain family member 14 cDNA (CARD14, BIMP2, CARMA2, PSORS2, GeneID 79092); mutated sequence
Discosoma sp. red fluorescent protein DNA (DsRed1); mutated monomeric variant (mCherry)
Promoter: Human elongation factor 1α promoter (EF1α)
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; preferably recombination-deficient strains
Mammalian cells
Parental clone: pLVX-EF1alpha-IRES-mCherry; pENTR3C-hCARMA2-H171N
Further information: The plasmid was constructed by isolating the human CARD14 coding sequence from pENTR3C-hCARMA2-H171N as an EcoRI/XbaI fragment, and cloning it into the pLVX-EF1alpha-IRES-mCherry vector.
The human CARD14 coding sequence contains an H171N mutation.
pLVX-EF1alpha-hCARMA2-H171N-IRES-mCherry is an HIV-1-based, lentiviral expression vector that allows the simultaneous expression of mutated human CARD14 and mCherry in any mammalian cell type, including primary cells.
The plasmid contains all viral processing elements necessary for the production of replication-incompetent lentivirus, as well as elements to improve viral titer and transgene expression. 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. The Rev-response element (RRE) further increases viral titers by enhancing the transport of unspliced viral RNA out of the nucleus and the central polypurine tract/central termination sequence element (cPPT/CTS) creates a central DNA flap during target cell infection that increases nuclear import of the viral genome, resulting in improved vector integration and more efficient transduction.
The nucleotide sequence of the wild type human CARD14 coding sequence corresponds with the EMBL Nucleotide Sequence Database accession number AF322642.1.
EMBL Accession number: AF322642.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 28/02/2018
Authenticity test: The plasmid still needs to be subjected to the authenticity test.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Dr J. Staal(1) (2) and Prof. R. Beyaert(1) (2).
(1) VIB-UGent Center for Inflammation Research, VIB, 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: 28°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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