Last data update: 24 January 2024 16:39 CET
Plasmid name: pRMCE-DV1-hMALT1 (LMBP 9111)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p9111.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human MALT1 paracaspase cDNA (MALT1, MLT, PCASP1, paracaspase 1) Aequorea victoria green fluorescent protein DNA (GFP); enhanced red-shifted variant (EGFP) |
| Promoter: | Mouse phosphoglycerate kinase 1 promoter (PGK1) Escherichia coli lac operon promoter |
| Ribosome binding site: |
Internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV) polyprotein gene |
| Terminator: | Bovine growth hormone polyadenylation signal (BGH polyA) Simian virus 40 polyadenylation signal (SV40 polyA) |
| Selection marker: | Ampicillin (amp) |
| Replicon: | Escherichia coli plasmid pMB1 origin |
| Host range: | Escherichia coli Mammalian cells |
| Parental clone: | pRMCE DV1; pENTR221-hMALT1 |
| Further information: | This conditional Gateway expression plasmid was constructed by cloning the human MALT1 coding sequence from pENTR221-hMALT1 into the pRMCE DV1 vector via Gateway LR recombination. This plasmid is a highly efficient Recombinase-Mediated Cassette Exchange (RMCE)-compatible, conditional Gateway expression vector. The plasmid contains: - two heterospecific Saccharomyces cerevisiae 2μ FLP Recombinase Target sites (one wt and one mutant FRT site) - two bacteriophage P1 Cre recombinase target sites (loxP sites) - two adjacent copies of the chicken β-globin 5' DNase I hypersensitive site 4 (5'HS4) insulator core sequence. Insulators are DNA sequences which possess the ability to protect expressing genes from inappropriate signals emanating from their surrounding environment by acting as barriers that prevent the advance of nearby condensed chromatin that may otherwise silence expression. - a splice acceptor (SA) for linking the cDNA of interest to the endogenous ROSA26 promoter. This Gateway expression vector, by co-transfection of a FlpE plasmid, can be targeted to the ROSA26 locus of G4 ROSALUC ES cells using a trap-coupled RMCE approach, hereby restoring neomycin resistance. Cre-mediated removal of the loxP-flanked stop cassette leads to ROSA26-based constitutive expression of the inserted cDNA. The G4 ROSALUC ES cells (LMBP 10507CB) are available at BCCM/GeneCorner as well. 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 pRMCE-DV1-hMALT1-WT. |
| EMBL Accession number: | AF130356.2, view at EMBL, GenBank, DDBJ |
| 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: | - |
| Related plasmid reference: | Haenebalcke et al., Stem Cell Rev. Rep. 9 (2013), 774-785 [PMID: 23877658] [DOI: 10.1007/s12015-013-9458-z] |
| 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: |
pRMCE-DV1-hMALT1 (LMBP 9111) 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.