Last data update: 05 December 2020 04:15 CET
Plasmid name: pCAGGS-E-hCASP-2 (LMBP 4716)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner core plasmid|
(View with Genome Compiler)
Human cysteinyl aspartate specific proteinase 2 cDNA (caspase-2, CASP-2, CASP2, NEDD2, ICH1)|
|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 was constructed by inserting a NotI-BglII PCR fragment, containing the hCASP-2 coding sequence, between the NotI and BglII site of pCAGGS-Etag-INCA.
pCAGGS-E-hCASP-2 is useful for highly efficient expression of hCASP-2, fused in phase to the N-terminal E-tag, under the control of the AG promoter and the hCMV-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.
Other name of the plasmid is pCAGGS-E-hCASPASE-2.
|EMBL Accession number:||-|
|Latest sequence update:||06/11/2003|
Nucleotide sequence at the start of the fusion gene: ---------------------- E-tag ---------------------> 5' ... TTCCACC.ATG.GGT.GCG.CCG.GTG.CCG.TAT.CCG.GAC.CCG.CTG.GAA.CCG.CGT.GCG. Met Gly Ala Pro Val Pro Tyr Pro Asp Pro Leu Glu Pro Arg Ala *** NotI NcoI StyI ----------------------- hCASP-2 -----------------------> GCC.GCC.ATG.GCC.GCT.GAC.AGG.GGA.CGC.AGG.ATA.TTG.GGA.GTG.TGT.GGC ... 3' Ala Ala Met Ala Ala Asp Arg Gly Arg Arg Ile Leu Gly Val Cys Gly BglI *** SphI NcoI StyI Nucleotide sequence at the end of the fusion protein: ----------------- hCASP-2 -----------------> -- ß-globin polyA -> 5' ... CTC.TAC.CTG.TTC.CCA.GGA.CAC.CCT.CCC.ACA.TGA.AGATCTTTTTCCCTCTGCC ... 3' Leu Tyr Leu Phe Pro Gly His Pro Pro Thr +++ BglII ***: start codon. +++: termination codon. Punctuation indicates reading frame.
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: BamHI/XmnI, BglII/NotI, EcoRI, HindIII/SpeI and XbaI.|
|History of deposit:||This plasmid was deposited by M. Lamkanfi(1) (2), Dr M. Kalai(1) (2) and Prof. Dr P. Vandenabeele(1) (2). It was constructed by M. Lamkanfi(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 MC1061(λ)|
|Host reference:||Mertens et al., Gene 164 (1995), 9-15 [PMID: 7590329]
|Related host reference:||Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493]
|Cultivation medium:||LB-Lennox + ampicillin (100 μg/ml)|
|Other culture collection numbers:||-|
Refer in your Materials and Methods:
|pCAGGS-E-hCASP-2 (LMBP 4716) is available at BCCM/GeneCorner. This plasmid was deposited by M. Lamkanfi , Dr M. Kalai and 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.