Last data update: 25 October 2020 04:11 CET
Plasmid name: pCAGGS-E-exoT-GAPmin (LMBP 4884)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner non-core plasmid|
|Cloned DNA:||Pseudomonas aeruginosa PAO1 exoenzyme T cDNA (exoT); with mutation in the GAP domain
|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:||pCAGGS-E-exoT-GAPmin is useful for highly efficient expression of mutated exoT, lacking GAP activity, under control of the AG promoter and the hCMV-IE enhancer in various mammalian cells.
As compared to the wt exoT cDNA, the arginine (R) codon at position 149 was replaced by a lysine (K) codon, abolishing a PvuI site.
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 the wt exoT cDNA corresponds with the Genbank accession number NC_002516.2.
Other name of the plasmid is pCAGGSE-ExoT GAP-.
|EMBL Accession number:||NC_002516.2, view at EMBL, GenBank, DDBJ|
|Latest sequence update:||03/09/2008|
|Authenticity test:||The plasmid still needs to be subjected to the authenticity test.|
|History of deposit:||This plasmid was deposited by Prof. Dr R. Beyaert(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
|Related plasmid reference:||Galle et al., J. Cell. Mol. Med. 12 (2008), 1767-1776 [PMID: 18081695]
|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)|
|Other culture collection numbers:||-|
Refer in your Materials and Methods:
|pCAGGS-E-exoT-GAPmin (LMBP 4884) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr R. Beyaert .|
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.