Last data update: 24 January 2024 16:39 CET
Plasmid name: pBKEF-NLScre (LMBP 4477)
New search | Print data sheet |
Price category: | Cat. 1 (cf. price list) |
Status: | GeneCorner non-core plasmid |
Depositor's sequence: | p4477.gb |
Sequence analysis results Genecorner: |
- |
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) |
Ribosome binding site: |
- |
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. |
Class: | Recombinant plasmid |
Type: | Plasmid |
History of deposit: | This plasmid was deposited by Prof. Dr W. Muller(1). (1) Faculty of Life Sciences, University of Manchester, Manchester, United Kingdom |
Plasmid reference: | - |
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) |
Cultivation temperature: | 37°C |
Biosafety level: | L1 |
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.