Last data update: 24 January 2024 16:39 CET
Plasmid name: pUC19-KpnI-BamHI-CaspM1-EcoRI-stop-ApaI (LMBP 9794)
| 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, GeneID 10892); mutated sequence |
| Promoter: | Escherichia coli lac operon promoter |
| Ribosome binding site: |
Ribosome binding site (RBS) of the Escherichia coli lac Z gene (lacZ) |
| Terminator: | - |
| Selection marker: | Ampicillin (amp) |
| Replicon: | Escherichia coli plasmid pMB1 origin |
| Host range: | Escherichia coli |
| Parental clone: | pUC19 |
| Further information: | The plasmid was constructed by cloning the synthetic human MALT1 coding sequence into the pUC19 vector. The human MALT1 coding sequence contains the following mutations: R149D, L539R, D462Q, A498S, E500R, Q502G. This leads to a caspase version of MALT1 (CaspM). Additionally, the internal EcoRI site has been deleted. R149D is an autoprocessing mutation. L539R, D462Q, A498S, E500R, Q502G are active site mutations predicted by modelling to switch MALT1 from a paracaspase to a caspase. The purpose of this mutant is 2-fold: * Proof-of-concept switching between paracaspase and caspase * For screening of novel inhibitors - using survival as inhibition marker could simultaneously select against inhibitors with poor inhibition capacity in cells and toxic compounds. |
| EMBL Accession number: | - |
| Latest sequence update: | 28/10/2019 |
| 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 Prof. Dr R. Beyaert(1) (2) and Dr J. Staal(1) (2). (1) 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: | 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: |
pUC19-KpnI-BamHI-CaspM1-EcoRI-stop-ApaI (LMBP 9794) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr R. Beyaert and Dr J. Staal . |
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.