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GeneCorner plasmid details

Last data update: 24 January 2024 16:39 CET

Plasmid name: pEF6-hMALT1-P2A-Hyg (LMBP 9271)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p9271.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Human MALT1 paracaspase cDNA (MALT1, MLT, PCASP1, paracaspase 1)
Porcine teschovirus-1 2A peptide (P2A)
Promoter: Human elongation factor 1α promoter (EF1α)
Phage T7 gene 10 promoter (T7g10)
Escherichia coli lac operon promoter
Ribosome
binding site:
-
Terminator: Simian virus 40 polyadenylation signal (SV40 polyA)
Selection marker: Hygromycin (hyg); in-frame translation
Ampicillin (amp)
Replicon: Escherichia coli plasmid pMB1 origin
Host range: Escherichia coli
Mammalian cells
Parental clone: pEF6-P2A-Hyg; pCD-MK
Further information: The plasmid was constructed by PCR amplifying the human MALT1 coding sequence from pCD-MK and cloning it in the KpnI/SpeI opened pEF6-P2A-Hyg vector.
P2A is a self-cleaving peptide, fulfilling the same role as an IRES, but has a much shorter sequence.
The purpose of this plasmid is to enforce expression of the human MALT1 mutant in selection-resistant cells by expressing the antibiotic resistance marker as an in-frame translation with your gene of interest, linked by the P2A autoprocessing peptide.
The parental pEF6-P2A-Hyg vector lacks part of the pEF6 backbone, resulting in a deletion of among others the SV40 origin of replication. As a result, this plasmid is only suitable for stable expression after random genomic integration of the linearised plasmid.
The nucleotide sequence of the human MALT1 coding sequence corresponds with the EMBL Nucleotide Sequence Database accession number AF130356.2.
Other name of the plasmid is pEF6-hMALT1-WT-P2A-Hyg.
EMBL Accession number: AF130356.2, view at EMBL, GenBank, DDBJ
Latest sequence update: 12/04/2017
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: Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660]
Rodriguez-Quinones et al., Focus 15 (1993), 110-112
Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791] [DOI: 10.1016/s0022-2836(83)80284-8]
Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187]
Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051]
Cultivation medium: LB-Lennox + ampicillin (100 μg/ml)
Cultivation temperature: 37°C
Biosafety level: L1
Other culture collection numbers: -

Refer in your Materials and Methods:

pEF6-hMALT1-P2A-Hyg (LMBP 9271) is available at BCCM/GeneCorner. The plasmid was deposited by Dr J. Staal and Prof. Dr R. Beyaert.

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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