Last data update: 24 November 2020 04:18 CET
Plasmid name: pPNhANCTN (LMBP 2938)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner non-core plasmid|
|Cloned DNA:||Human αN-catenin cDNA (aNctn, CAP-R, CTNNA2, catna2)|
|Promoter:||Mouse phosphoglycerate kinase 1 promoter (PGK1)
Escherichia coli lac operon promoter
Phage T3 promoter
|Terminator:||Herpes simplex virus (HSV) thymidine kinase polyadenylation signal (TK polyA)
Mouse phosphoglycerate kinase 1 polyadenylation signal (PGK1 polyA)
|Selection marker:||Ampicillin (amp)
Neomycin (neo; G418)
|Replicon:||Escherichia coli plasmid pMB1/ColE1 origin|
|Host range:||Escherichia coli; preferably recombination-deficient strains
|Parental clone:||pPNT; phKALfhANCTN; pBluhANCTNf|
|Further information:||This plasmid was constructed by a four-point-ligation: 1) a BstEII-KpnI vector fragment from pPNT, 2) a KpnI-PstI (nucleotide position 2684) fragment from pPNT, containing the mouse PGK1 promoter, 3) a PstI-NcoI fragment from pBluhANCTNf, containing part of the human αN-catenin gene, and 4) an NcoI-BstEII fragment from phKALfhANCTN, containing the completing part of the human αN-catenin gene.
This plasmid can be used for expression of human αN-catenin in mammalian cells.
The neomycin resistance gene contains some modifications at the start of the coding sequence as compared to the wild type gene.
As the plasmid contains the mouse PGK1 polyA and promoter sequence twice, one should be aware of the possibility of deletions due to homologous recombination.
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 names of the plasmid are pPNαNCAT and pPNHANCAT.
|EMBL Accession number:||-|
|Latest sequence update:||21/10/2002|
|Authenticity test:||The plasmid still needs to be subjected to the authenticity test.|
|History of deposit:||This plasmid was deposited by Prof. Dr F. Van Roy(1) (2). It was constructed by F. Nollet(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
|Related plasmid reference:||Tybulewicz et al., Cell 65 (1991), 1153-1163 [PMID: 2065352]
|Restricted distribution:||- BCCM MTA|
|Distributed as:||H/P active culture or plasmid DNA|
|Host for distribution:||Escherichia coli K12 DH5αF(-)|
|Cultivation medium:||LB-Lennox + ampicillin (100 μg/ml)|
|Other culture collection numbers:||-|
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.