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GeneCorner plasmid details

Last data update: 29 November 2020 04:28 CET

Plasmid name: pBlue-loxP-PGK-gb2-hyg-loxP (LMBP 6665)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p6665.gb (View with Genome Compiler)
p6665.txt
p6665.pdf
Sequence
analysis results
Genecorner:

-

Cloned DNA: -
Promoter: Mouse phosphoglycerate kinase 1 promoter (PGK1)
Synthetic prokaryotic EM7 promoter
Phage T7 gene 10 promoter (T7g10)
Phage T3 promoter
Escherichia coli lac operon promoter
Ribosome
binding site:
Ribosome binding site (RBS) of the Escherichia coli lac Z gene (lacZ)
Terminator: Synthetic polyadenylation signal (polyA)
Selection marker: Ampicillin (amp)
Hygromycin (hyg)
Replicon: Escherichia coli plasmid pMB1 origin
Phage f1 origin
Host range: Escherichia coli
Mammalian cells
Parental clone: pBluescriptII KS+
Further information: The plasmid was constructed by ligating the loxP-PGK-gb2-hyg-loxP PCR-cassette from Gene Bridges into the NotI/XhoI-opened pBluescriptII KS+ vector.
The plasmid contains two bacteriophage P1 Cre recombinase target sites (loxP sites) for excision of the cassette by Cre-recombinase.
The plasmid allows hygromycin selection in prokaryotic and eukaryotic cells.
When cloning a fragment downstream from the lac/tac/trc/N25/O2 promoter it may be advisable to use lacIq 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 LT726838.1.
The nucleotide sequence of the parental pBluescriptII KS+ vector corresponds with the EMBL Nucleotide Sequence Database accession number X52327.1.
EMBL Accession number: X52327.1, view at EMBL, GenBank, DDBJ
LT726838.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 23/04/2010
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BglI/EcoRI, NcoI and NotI/XhoI.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by I. Kahr and Prof. Dr F. Van Roy(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: PhD thesis Jäckel er al. (2017)
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 DH5α
Host reference: Focus 8 (1986), 9
Related host reference: Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051]
Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187]
Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791]
Rodriguez-Quinones et al., Focus 15 (1993), 110-112
Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660]
Cultivation medium: LB-Lennox + ampicillin (100 μg/ml)
Cultivation temperature: 37°C
Biosafety level: L1
Cultivation remark: -
Other culture collection numbers: -
Related website: http://www.genebridges.com/selection-cassettes/A011

Refer in your Materials and Methods:

pBlue-loxP-PGK-gb2-hyg-loxP (LMBP 6665) is available at BCCM/GeneCorner. This plasmid was deposited by I. Kahr and Prof. Dr F. Van Roy .

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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