Last data update: 03 December 2020 04:26 CET
Plasmid name: pAS2mCASP-11m (LMBP 3922)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner non-core plasmid|
|Cloned DNA:||Mouse cysteinyl aspartate specific proteinase 11 cDNA (caspase-11, CASP-11, Ich3, Casp4); mutated sequence
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
|Terminator:||Saccharomyces cerevisiae alcohol dehydrogenase terminator (ADH1)
Saccharomyces cerevisiae iso-1-cytochrome C terminator (CYC1)
Saccharomyces cerevisiae 2 micron plasmid (2μ) FLP terminator
|Selection marker:||Ampicillin (amp)
|Replicon:||Escherichia coli plasmid pMB1 origin
Phage f1 origin
Saccharomyces cerevisiae 2 micron plasmid origin (2μ); incl. region conferring stability (STB)
|Host range:||Escherichia coli
Saccharomyces cerevisiae; trp1(-)
|Further information:||The plasmid was constructed as follows: the plasmid pAS2 was opened with BalI(MscI)/SalI and the mutated mouse cysteinyl aspartate specific proteinase 11 cDNA (caspase-11, mCASP-11) was inserted by the use of primers.
mCASP-11 contains a C254S mutation. The cysteine codon at position 254 was replaced by a serine codon, creating an additional NheI site.
The nucleotide sequence of the pAS2 vector corresponds with the EMBL Nucleotide Sequence Database accession number U30496.1, but adapted as follows: 1) the 'N' nucleotide was removed, restoring the coding sequence of cycloheximide; 2) the sequence GATTTC between GALbd and the HA epitope was replaced by GAATTC, an EcoRI site that has been experimentally confirmed in pAS2; 3) the sequence TTCGAA (Csp45I site) in the ADH1 promoter was replaced by TCGAAG, eliminating the Csp45I site that could not be experimentally confirmed in pAS2.
Other name of the plasmid is pAS2PROCASP-11M.
|EMBL Accession number:||U30496.1, view at EMBL, GenBank, DDBJ|
|Latest sequence update:||06/02/2004|
Nucleotide sequence around the HA epitope and the mutated mouse CASP-11 coding sequence: GALbd -> 147 ----------- HA epitope -----------> 5' ... TCG.CCG.GAA.TTC.ATG.GCT.TAC.CCA.TAC.GAT.GTT.CCA.GAT.TAC.GCT.AGC.TTG.GGT.GGT. Ser Pro Glu Phe Met Ala Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Ser Leu Gly Gly EcoRI NheI ------------------------ mCASP-11m ------------------------> 1 254 373 CAT.ATG.GCC.ACC.ACC.ATG.GCT.GAA.AAC.AAA ... GCT.AGC.AGA ... TTC.TTT.CCT.GGC.AAC. His Met Ala Thr Thr Met Ala Glu Asn Lys Ala Ser Arg Phe Phe Pro Gly Asn BalI NcoI NheI StyI ^ ^ GTC.GAC.CTG.CAG.CCA.AGC.TAA.TTCCGGGCG ... 3' Val Asp Leu Gln Pro Ser +++ SalI PstI HindII ^ : Mutated nucleotide. +++: Termination codon. Punctuation indicates reading frame.
|Authenticity test:||The plasmid still needs to be subjected to the authenticity test.|
|History of deposit:||This plasmid was deposited by Prof. Dr R. Beyaert(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
|Related plasmid reference:||Wang et al., Cell 92 (1998), 501-509 [PMID: 9491891]
|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)|
|Other culture collection numbers:||-|
Refer in your Materials and Methods:
|pAS2mCASP-11m (LMBP 3922) is available at BCCM/GeneCorner. This 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.