Last data update: 29 November 2020 04:28 CET
Plasmid name: pBKEF-NLScre (LMBP 4477)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner non-core plasmid|
|Cloned DNA:||Phage P1 cre gene encoding recombinase
SV40 nuclear localization signal (NLS); N-terminal
|Promoter:||Human elongation factor 1α promoter (EF1α)
Escherichia coli β-lactamase promoter (amp)
Simian virus 40 early promoter (SV40 early)
|Terminator:||Human granulocyte macrophage colony stimulating factor polyadenylation signal (GM-CSF polyA)
Herpes simplex virus (HSV) thymidine kinase polyadenylation signal (TK polyA)
|Selection marker:||Neomycin (neo; G418; kanamycin (kan))|
|Replicon:||Escherichia coli plasmid pMB1 origin
Simian virus 40 bidirectional origin (SV40)
Phage f1 origin
|Host range:||Escherichia coli
Mammalian cells; SV40 permissive cells
|Parental clone:||pBK-CMV; pEF-BOS; pGBF-NLScre|
|Further information:||The plasmid was constructed by inserting the phage P1 cre gene encoding recombinase (with N-terminal SV40 nuclear localization signal (NLS)) as a NheI/SpeI fragment of pGBF-NLScre between the XbaI sites of pBKEF-8.
pBKEF-8 was derived from pBK-CMV (Stratagene) by replacing the human CMV promoter and the SV40 polyA by the human EF1α promoter and the human GM-CSF polyA from pEF-BOS.
To generate stably transfected cells, use the NotI linearised plasmid.
pBKEF-NLScre has been successfully used to delete a loxP flanked neomycin resistance gene in ES cells. Still, if one is intending to perform partial deletions of three loxP sites in ES cells, it is recommended to try pBKEF-Kozakcre first, since NLScre might preferentially lead to complete deletions.
The nucleotide sequence of the cre gene corresponds with the EMBL Nucleotide Sequence Database accession number X03453.1.
|EMBL Accession number:||X03453.1, view at EMBL, GenBank, DDBJ|
|Latest sequence update:||15/05/2002|
|Authenticity test:||The plasmid still needs to be subjected to the authenticity test.|
|History of deposit:||This plasmid was deposited by Prof. Dr W. Muller(1).
(1) Faculty of Life Sciences, University of Manchester, Manchester, United Kingdom
|Restricted distribution:||- BCCM MTA|
|Distributed as:||H/P active culture or plasmid DNA|
|Host for distribution:||Escherichia coli K12 XL1-Blue MRF'|
|Host reference:||Jerpseth et al., Strategies 5 (1992), 81-83
|Cultivation medium:||LB-Lennox + kanamycin (25 μg/ml)|
|Other culture collection numbers:||-|
Refer in your Materials and Methods:
|pBKEF-NLScre (LMBP 4477) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr W. Muller.|
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.