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LMBP plasmid details

Last data update: 24 November 2020 04:18 CET

Plasmid name: pPNTNeoflox-msGCA1 (LMBP 4472)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p4472.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Mouse guanylate cyclase 1, soluble, alpha 3 cDNA (Gucy1a3, alfa 1 sGC, sGC-alfa1, sGCA1); homology arms
Promoter: Mouse phosphoglycerate kinase 1 promoter (PGK1)
Escherichia coli lac operon promoter
Phage T3 promoter
Ribosome
binding site:
-
Terminator: Herpes simplex virus (HSV) thymidine kinase polyadenylation signal (TK polyA)
Mouse phosphoglycerate kinase 1 polyadenylation signal (PGK1 polyA)
Selection marker: Ampicillin (amp)
Neomycin (neo; G418)
Thymidine kinase (TK)
Replicon: Escherichia coli plasmid pMB1/ColE1 origin
Host range: Escherichia coli; preferably recombination-deficient strains
Mammalian cells
Parental clone: pPNT
Further information: This plasmid was developed for 'knock-out' experiments in mammalian cells by double homologous recombination (positive and negative selection).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.
EMBL Accession number: -
Latest sequence update: 16/05/2002
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BglI, NotI/XhoI and SmaI.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Prof. Dr P. Brouckaert(1) (2). It was constructed by M. Buys(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: -
Restricted distribution: - VIB/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: -

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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