Last data update: 24 January 2024 16:39 CET
Plasmid name: pAS3mCASP8p20C/S (LMBP 4309)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p4309.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Mouse cysteinyl aspartate specific proteinase 8 cDNA (caspase-8, CASP-8, Casp8, MCH5, MACH, FLICE) Saccharomyces cerevisiae GAL4 DNA-binding domain (GALbd) Influenza HA epitope encoding the haemagglutinin tagging peptide; N-terminal |
| Promoter: | Saccharomyces cerevisiae alcohol dehydrogenase promoter (ADH1) Escherichia coli lac operon promoter |
| Ribosome binding site: |
- |
| Terminator: | Saccharomyces cerevisiae alcohol dehydrogenase terminator (ADH1) Saccharomyces cerevisiae 2 micron plasmid (2μ) FLP terminator |
| Selection marker: | Ampicillin (amp) ADE2; auxotrophic |
| Replicon: | Escherichia coli plasmid pMB1 origin Saccharomyces cerevisiae 2 micron plasmid origin (2μ); incl. region conferring stability (STB) |
| Host range: | Escherichia coli Saccharomyces cerevisiae; ade2(-) |
| Parental clone: | pAS3 |
| Further information: | The plasmid was created by PCR amplifying the mCASP8p20C/S cds fragment and ligating it as an NcoI-SalI fragment into the NcoI-SalI opened pAS3 vector. The mCASP8 cds was mutated via PCR, replacing the cystein in the catalytic site by serin. pAS3 is a yeast two-hybrid vector expressing the cloned mCASP8 gene as a fusion protein with GAL4DB. The vector is derived from the pAS2 vector containing the ade2 gene for auxotrophic selection in yeasts. |
| EMBL Accession number: | - |
| Latest sequence update: | 21/04/2009 |
| Sequence detail: | Primers used for PCR amplification and mutation of the mCASP8 cds:
Forward: 5' CC.GCC.ATG.GCC.AGT.GAG.TCA.CGG.ACT.TCA.GAC.A
NcoI
Reverse: 5' GCGGGTCGAC.TCA.ATC.AGG.CAC.TCC.TTT.CTG.GAA.GTT.ACT.TCC.TTG.CGC.AGC.CTG.AAT.GA
SalI +++ ***
+++: Termination codon of GAL4DB-mCASP8 fusion protein
***: mutated nucleotides |
| 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 P. Vandenabeele(1) (2). (1) Department for Molecular Biomedical 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 MC1061 |
| Host reference: | Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493] |
| Related host reference: | Brigé et al., Biochem. J. 394 (2006), 335-344 [PMID: 16293111] |
| 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: |
pAS3mCASP8p20C/S (LMBP 4309) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr P. Vandenabeele . |
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.