Last data update: 24 November 2020 04:18 CET
Plasmid name: pPHT (LMBP 3960)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner core plasmid|
(View with Genome Compiler)
|Promoter:||Mouse phosphoglycerate kinase 1 promoter (PGK1)
Escherichia coli lac operon promoter
Phage T3 promoter
|Terminator:||Mouse phosphoglycerate kinase 1 polyadenylation signal (PGK1 polyA)
Herpes simplex virus (HSV) thymidine kinase polyadenylation signal (TK polyA)
|Selection marker:||Ampicillin (amp)
Thymidine kinase (TK)
|Replicon:||Escherichia coli plasmid ColE1 origin|
|Host range:||Escherichia coli; preferably recombination-deficient strains
|Parental clone:||pPNT; pSP72hyg|
|Further information:||The plasmid was constructed by inserting an XhoI-XbaI PCR fragment of pSP72hyg, containing the hygromycin resistance gene, between the XhoI and XbaI sites of pPNT, replacing the neomycin resistance gene.
This plasmid was developed for gene 'knock-out' experiments in mammalian cells by double homologous recombination (positive and negative selection).
The plasmid can be used to make stable cell lines.
As the plasmid contains the mouse PGK1 promoter and the mouse PGK1 polyA 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.
The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727296.1.
The XhoI-XbaI hygromycin insert has been sequenced by BCCM/GeneCorner.
|EMBL Accession number:||LT727296.1, view at EMBL, GenBank, DDBJ|
|Latest sequence update:||24/10/2008|
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: EcoRI, HindIII, NotI, NotI/NruI, NotI/XbaI and SmaI.|
|History of deposit:||This plasmid was deposited by Dr R. De Vos(1). It was constructed by Dr J. Tavernier(1).
(1) Roche Research Gent, 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 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:||LMBP 02823|
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.