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LMBP plasmid details

Last data update: 24 November 2020 04:18 CET

Plasmid name: pPLc28BHTNFs (LMBP 1708)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p1708.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Ampicillin resistance gene (amp); signal sequence
Human tumor necrosis factor cDNA (TNF); mature sequence
Promoter: Escherichia coli β-lactamase promoter (amp)
Phage λ major leftward promoter (λ PL)
Ribosome
binding site:
Ribosome binding site (RBS) of the Escherichia coli ampicillin resistance gene (amp)
Terminator: -
Selection marker: Ampicillin (amp)
Replicon: Escherichia coli plasmid pMB1 origin
Host range: Escherichia coli; use strains with a cI function, cIts for PL controlled expression
Parental clone: pPLc2820BA; pPLcmuHTNF1
Further information: This plasmid was constructed as follows : to the ApaI site (blunted by T4 polymerase exonuclease treatment) of pPLc2820BA, an AvaI linker was ligated, and the AvaI-HindIII fragment of pPLcmuHTNF1 (containing the mature HTNF gene) was inserted.
The plasmid contains the AMP signal sequence, fused in frame with the mature human TNF coding sequence.
Strains, containing pPLc28BHTNF secrete mature human TNF into the periplasmic space.
The ampicillin resistance gene does not contain a PstI restriction site.
Expresssion in λ wt lysogen : constitutive synthesis under control of the amp promoter at any physiological temperature.
Expression in cIts : λ-PL induction at 42°C.
The first 7 amino acids of the mature human TNF are identical to wild type, but are encoded by a synthetic linker sequence (nucleotide sequence different from wild type human TNF)
Other name of the plasmid is pPLc2820BTNF.
EMBL Accession number: -
Latest sequence update: 29/08/1990
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 E. Remaut(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: Leemans et al., Gene 85 (1989), 99-108 [PMID: 2695405]
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 MC1061(λ)
Host reference: Mertens et al., Gene 164 (1995), 9-15 [PMID: 7590329]
Related host reference: Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493]
Cultivation medium: LB-Lennox + ampicillin (100 μg/ml)
Cultivation temperature: 28°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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