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GeneCorner plasmid details

Last data update: 24 January 2024 16:39 CET

Plasmid name: pBKEF-NLScre (LMBP 4477)

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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.

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