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LMBP plasmid details

Last data update: 24 November 2020 04:18 CET

Plasmid name: pPNT (LMBP 2682)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p2682.gb (View with Genome Compiler)
p2682.txt
p2682.pdf
Sequence
analysis results
Genecorner:

-

Cloned DNA: -
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 ColE1 origin
Host range: Escherichia coli; preferably recombination-deficient strains
Mammalian cells
Parental clone: pUCNot
Further information: This plasmid was developed as a vector for gene 'knock-out' experiments in mammalian cells by double homologous recombination (positive and negative selection).
The neomycin resistance gene contains some modifications at the start of the coding sequence as compared to the wild type gene.
As the plasmid contains the mouse PGK1 promoter and the mouse PGK1 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.
The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727535.1.
The nucleotide sequence of this plasmid was compiled based on the publications mentioned in 'Plasmid reference' and 'Related reference'.
Unpublished sequences and linkers have been sequenced at the Department of Biomedical Molecular Biology, Ghent University, Belgium.
The linker region between the 3' UTR of the mouse PGK1 gene and the mouse PGK1 promoter has been sequenced at Roche Research Ghent.
EMBL Accession number: LT727535.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 22/11/2006
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: EcoRI, FspI, SmaI and SpeI.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Dr R.C. Mulligan(1).
(1) Whitehead Institute for Biomedical Research, Massachusetts Institute of Technology, Cambridge, USA
Plasmid reference: Tybulewicz et al., Cell 65 (1991), 1153-1163 [PMID: 2065352]
Related plasmid reference: Thomas et al., Cell 51 (1987), 503-512 [PMID: 2822260]
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 MC1061
Host reference: Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493]
Related host reference: Brigé et al., Biochem. J. 394 (2006), 335-344 [PMID: 16293111]
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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