Last data update: 24 January 2024 16:39 CET
Plasmid name: pEF6-hMALT1-C464A-P2A-Hyg (LMBP 9272)
New search | Print data sheet |
Price category: | Cat. 1 (cf. price list) |
Status: | GeneCorner non-core plasmid |
Depositor's sequence: | not available |
Sequence analysis results Genecorner: |
- |
Cloned DNA: |
Human MALT1 paracaspase cDNA (MALT1, MLT, PCASP1, paracaspase 1); mutated sequence 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: | Bovine growth hormone polyadenylation signal (BGH 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-p20mut |
Further information: | The plasmid was constructed by PCR amplifying the mutant human MALT1 coding sequence from pCD-MK-p20mut and cloning it in the KpnI/SpeI opened pEF6-P2A-Hyg vector. As compared to the wt hMLT cDNA, the predicted catalytic cysteine (C) codon at position 464, in the caspase(p20)-like active site of human MALT1, was replaced by an alanine (A) codon, creating an extra PauI site. 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 wild type human MALT1 coding sequence corresponds with the EMBL Nucleotide Sequence Database accession number AF130356.2. |
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-C464A-P2A-Hyg (LMBP 9272) 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.