Last data update: 24 January 2024 16:39 CET
Plasmid name: pET-23b-hCASP6-D23A-D193A-His (LMBP 9093)
| 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 caspase 6 cDNA (CASP6, MCH2, GeneID 839); mutated sequence Histidine tag (His-tag); C-terminal |
| Promoter: | Phage T7 gene 10 promoter (T7g10) |
| Ribosome binding site: |
Ribosome binding site (RBS) of the phage T7 gene 10 (T7g10) |
| Terminator: | Phage T7 gene 10 terminator (T7g10) |
| Selection marker: | Ampicillin (amp) |
| Replicon: | Escherichia coli plasmid pMB1 origin Phage f1 origin |
| Host range: | Escherichia coli |
| Parental clone: | Caspase-6 pET23b 6xHis |
| Further information: | The plasmid was constructed by introducing a D23A and a D193A mutation into the human CASP6 coding sequence of Caspase-6 pET23b 6xHis via site-directed mutagenesis. The D23A and D193A mutations in the human CASP6 coding sequence prevent autoprocessing by respectively making the pro-domain and the auto-processing site uncleavable. This mutant expresses as a single chain zymogen but gains activation upon processing with human CASP3. This plasmid contains the T7 gene 10 promoter whose transcription is initiated by T7 RNA polymerase, making the vector useful for in vitro preparation of single-stranded RNAs from the cloned human CASP6 gene. Other name of the plasmid is Caspase-6 6xHis D23AD193A pET23b. |
| EMBL Accession number: | - |
| Latest sequence update: | 05/04/2017 |
| 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 Prof. Dr R. Beyaert(1)(2). It was constructed by Dr. B. Van Raam(3) (1) Center for Inflammation Research, VIB, Ghent, Belgium (2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium (3) BioMed X Innovation Center, Heidelberg, Germany |
| Plasmid reference: | - |
| Related plasmid reference: | Edgington et al., Chem. Biol. 19 (2012), 340-352 [PMID: 22444589] [DOI: 10.1016/j.chembiol.2011.12.021] |
| 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 |
| Cultivation remark: | - |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pET-23b-hCASP6-D23A-D193A-His (LMBP 9093) is available at BCCM/GeneCorner. The plasmid was deposited by 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.