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LMBP plasmid details

Last data update: 24 November 2020 04:18 CET

Plasmid name: pPNhANCTN (LMBP 2938)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p2938.gb
Sequence
analysis results
Genecorner:

-

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
Ribosome
binding site:
-
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
Mammalian cells
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.
Class: Recombinant plasmid
Type: Plasmid
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
Plasmid reference: -
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(-)
Host reference: -
Cultivation medium: LB-Lennox + ampicillin (100 μg/ml)
Cultivation temperature: 37°C
Biosafety level: L1
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.

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