Last data update: 30 October 2020 04:11 CET
Plasmid name: pCAGGS-hCASP-6 (LMBP 3831)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner core plasmid|
(View with Genome Compiler)
|Cloned DNA:||Human cysteinyl aspartate specific proteinase 6 cDNA (caspase-6, hCASP-6, Mch2)
|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
|Parental clone:||pCAGGS; pBluhCASP-6|
|Further information:||The plasmid was constructed by inserting the human caspase-6 cDNA as a XhoI fragment from pBluhCASP-6 (blunted with Pfu DNA polymerase) into the unique MscI (BalI) site of pCAGGS. The orientation of the inserted fragment is sense relative to the AG promoter.
pCAGGS-hCASP-6 is useful for highly efficient expression of human caspase-6 under the control of the chicken β-actin/rabbit β-globin hybrid promoter (AG) 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 LT727412.1.
The nucleotide sequence of hCASP-6 corresponds with the Genbank accession number U20536.1.
Other name of the plasmid is pcaggsmch#5.
|EMBL Accession number:||U20536.1, view at EMBL, GenBank, DDBJ
LT727412.1, view at EMBL, GenBank, DDBJ
|Latest sequence update:||28/01/2005|
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: AvaIII, BglII and NcoI.|
|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
|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:||Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051]
Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187]
Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791]
Rodriguez-Quinones et al., Focus 15 (1993), 110-112
Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660]
|Cultivation medium:||LB-Lennox + ampicillin (100 μg/ml)|
|Other culture collection numbers:||-|
Refer in your Materials and Methods:
|pCAGGS-hCASP-6 (LMBP 3831) 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.