Last data update: 24 January 2024 16:39 CET
Plasmid name: pCAGGS-EmCASP-1C285A (LMBP 4937)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p4937.gb
(View with Genome Compiler) p4937.txt p4937.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Mouse cysteinyl aspartate specific proteinase 1 cDNA (caspase-1, CASP-1, ICE, Casp1); C285A mutant 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 |
| Further information: | pCAGGS-EmCASP-1C285A codes for the enzymatically inactive mutant of mouse caspase-1. The mutation is described as C285A, according to the numbering of the codons in the human caspase-1 coding sequence, although the mutated mouse codon is located on position 284. The plasmid is useful for highly efficient expression of mouse CASP-1C285A, fused to the N-terminal E-tag, under the control of the AG promoter and the human CMV-IE enhancer in various mammalian cells. 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 LT727059.1. The nucleotide sequence of the wild type mouse CASP-1 cDNA corresponds with the EMBL Nucleotide Sequence Database accession number BC008152.1. Other name of the plasmid is pCAGGS-E-mCASP1 C/A. |
| EMBL Accession number: | BC008152.1, view at EMBL, GenBank, DDBJ LT727059.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 15/05/2008 |
| Sequence detail: | E-tag
---------------------------------------------------
5' ... GAATTCCACC ATG.GGT.GCG.CCG.GTG.CCG.TAT.CCG.GAC.CCG.CTG.GAA.CCG.CGT.GCG.GCC.
Met Gly Ala Pro Val Pro Tyr Pro Asp Pro Leu Glu Pro Arg Ala Ala
EcoRI BspMII NotI
|--> mouse CASP-1
| 2 3 4 280 284
GCT.GAC.AAG...GAC.AAG.CCC.AAG.GTG.ATC.ATT.ATT.CAG.GCA.GCC.CGT.GGA.GAG.AAA.CAA...
Asp Lys Pro Lys Val Ile Ile Ile Gln Ala Ala Arg Gly Glu Lys Gln
**
Q A A R G
*: mutated nucleotide.
Punctuation indicates reading frame. |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BglII/NotI, NcoI, SphI and XbaI. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by 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: | - |
| Related plasmid reference: | Lamkanfi et al., J. Biol. Chem. 279 (2004), 24785-24793 [PMID: 15039421] |
| 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) |
| Cultivation temperature: | 37°C |
| Biosafety level: | L1 |
| Cultivation remark: | - |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pCAGGS-EmCASP-1C285A (LMBP 4937) 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.