Last data update: 24 January 2024 16:39 CET
Plasmid name: pCAGGS-CrmA-T291R (LMBP 4577)
New search | Print data sheet |
Price category: | Cat. 1 (cf. price list) |
Status: | GeneCorner core plasmid |
GeneCorner sequence: |
p4577.gb
(View with Genome Compiler) p4577.txt p4577.pdf |
Sequence analysis results Genecorner: |
- |
Cloned DNA: |
Cowpox virus cytokine response modifier A cDNA (crmA); mutated sequence |
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: | pCAGGScrmA |
Further information: | pCAGGS-CrmA-T291R is useful for highly efficient expression of mutated crmA under the control of the AG promoter and the human CMV-IE enhancer in various mammalian cells. As compared to the wt crmA cDNA, the threonine (T) codon at position 291 was replaced by an arginine (R) codon, creating a unique NruI (Bsp68I) site and abolishing a PvuII site. The crmA pointmutant can no longer inhibit caspase activity and consequently apoptosis. 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 LT727106.1. The nucleotide sequence of the crmA cDNA corresponds with the EMBL Nucleotide Sequence Database accession number M14217.1. |
EMBL Accession number: | M14217.1, view at EMBL, GenBank, DDBJ LT727106.1, view at EMBL, GenBank, DDBJ |
Latest sequence update: | 26/02/2008 |
Sequence detail: | 285 290 291 295 300 wt CrmA: 5' ... GAT.GTC.AAT.GAA.GAG.TAT.ACA.GAA.GCA.GCT.GCA.GCA.ACT.TGT.GCG.CTG ... 3' Asp Val Asn Glu Glu Tyr Thr Glu Ala Ala Ala Ala Thr Cys Ala Leu PvuII PstI 285 290 291 295 300 crmA mutant: 5' ... GAT.GTC.AAT.GAA.GAG.TAT.CGC.GAA.GCG.GCT.GCA.GCA.ACT.TGT.GCG.CTG ... 3' Asp Val Asn Glu Glu Tyr Arg Glu Ala Ala Ala Ala Thr Cys Ala Leu ^^^ ^ Bsp98I PstI NruI ^: mutated nucleotide. Punctuation indicates reading frame. |
Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BamHI/PvuII, Bsp68I/NcoI, Bsu15I/HindIII, EcoRI, PstI/PvuI 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 G. Van Loo(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: | Vercammen et al., J. Exp. Med. 187 (1998), 1477-1485 [PMID: 9565639] Tewari et al., Cell 81 (1995), 801-809 [PMID: 7774019] |
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-CrmA-T291R (LMBP 4577) 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.