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LMBP plasmid details

Last data update: 24 November 2020 04:18 CET

Plasmid name: pPN1 (LMBP 2912)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: ppn1.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Transposon Tn5 neomycin phosphotransferase gene (neo)
Promoter: Mouse phosphoglycerate kinase 1 promoter (PGK1)
Escherichia coli lac operon promoter
Phage T3 promoter
Ribosome
binding site:
-
Terminator: Mouse phosphoglycerate kinase 1 polyadenylation signal (PGK1 polyA)
Selection marker: Ampicillin (amp)
Neomycin (neo; G418)
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 derived from pPNT digested with Eco47III and HindII, followed by blunt-blunt ligation; in this way, the herpes simplex virus thymidine kinase gene was deleted.
This plasmid can be used for expression of heterologous genes under the control of the mouse PGK1 promoter by cloning them into the unique BstEII site. pPN1 can also be used as a negative control (e.g. for mouse E-cadherin, in combination with plasmid pPNEcad).
The neomycin resistance gene contains some modifications at the start of the coding sequence as compared to the wild type gene.
Since the plasmid contains the mouse PGK1 promoter and polyA sequence twice, one should be aware of the possibility of deletions due to homologous recombination.
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.
Other name of the plasmid is pPN.
EMBL Accession number: -
Latest sequence update: 13/04/1994
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 K. Staes(1) (2) 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: -
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
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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