Last data update: 24 January 2024 16:39 CET
Plasmid name: pCAGGS-E-hCASP-2 (LMBP 4716)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p4716.gb
(View with Genome Compiler) p4716.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human cysteinyl aspartate specific proteinase 2 cDNA (caspase-2, CASP-2, CASP2, NEDD2, ICH1) E-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 |
| Ribosome binding site: |
- |
| 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-Etag-INCA |
| 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 |
| Sequence detail: | 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. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| 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 |
| Plasmid reference: | - |
| 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) |
| Cultivation temperature: | 28°C |
| Biosafety level: | L1 |
| Cultivation remark: | - |
| 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.