Last data update: 28 October 2020 04:17 CET
Plasmid name: pCAGGS-FLAG-hCASP-1-C285A (LMBP 7424)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner core plasmid|
(View with Genome Compiler)
|Cloned DNA:||Human apoptosis-related cysteine peptidase cDNA (caspase-1, CASP1, CASP-1, ICE, P45, IL1BC); catalytically inactive mutant
FLAG epitope tag; N-terminal
|Promoter:||Chicken β-actin/rabbit β-globin hybrid promoter (AG)
Human cytomegalovirus immediate early promoter (CMV-IE); enhancer only
Escherichia coli lac operon promoter
|Terminator:||Rabbit β-globin polyadenylation signal (β-globin polyA)
Simian virus 40 polyadenylation signal (SV40 polyA)
|Selection marker:||Ampicillin (amp)|
|Replicon:||Escherichia coli plasmid pMB1 origin
Simian virus 40 bidirectional origin (SV40)
|Host range:||Escherichia coli
Mammalian cells; SV40 permissive cells
|Further information:||The plasmid contains a C285A mutation, resulting in a catalytically inactive form of human CASP-1.
This plasmid is useful for highly efficient expression of this catalytically inactive human CASP-1 gene under the control of the AG promoter and the human CMV-IE enhancer in various mammalian cells.
The AG promoter sequence consists of the chicken β-actin promoter, the first exon and part of the first intron (that seems to have a strong enhancer-like activity) linked to a rabbit β-globin fragment, consisting of a 3' part of the second intron (inclusive a branch point which is required for normal splicing reactions) and a 5' part of the third exon.
When cloning a fragment downstream from the lac promoter it may be advisable to use lacI(q) strains in order to prevent fortuitous expression of a possibly noxious polypeptide.
The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT726859.1.
The parental plasmid pCAGGS was completely sequenced by BCCM/GeneCorner.
Name mentioned in Kersse et al. (2011) is pCAGGS-FLAG-procaspase-1 C285A.
Other name of the plasmid is pCAGGS-Flag-hcaspase-1 C/A.
|EMBL Accession number:||LT726859.1, view at EMBL, GenBank, DDBJ|
|Latest sequence update:||07/12/2011|
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: BglII/XhoI, EcoRI, HindIII/SacII and NcoI/ScaI.|
|History of deposit:||This plasmid was deposited by K. Kersse(1) (2) and 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:||Kersse et al., J. Biol. Chem. 286 (2011), 35874-35882 [PMID: 21862576] [DOI: 10.1074/jbc.M111.242321]
|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:
|pCAGGS-FLAG-hCASP-1-C285A (LMBP 7424) is available at BCCM/GeneCorner. This plasmid was deposited by K. Kersse and Prof. Dr P. Vandenabeele and was published in Kersse et al., 2011.|
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.