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LMBP plasmid details

Last data update: 24 November 2020 04:18 CET

Plasmid name: pPLc28BHTNFm28Fs (LMBP 1799)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p1799.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Ampicillin resistance gene (amp); signal sequence
Human tumor necrosis factor cDNA (TNF); mutated 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: pPLc28BHTNF; pMaT4HTNFTm28F
Further information: The plasmid was constructed by inserting the AvaI-BstEII fragment of pMaT4HTNFTm28F, containing the
mutated mature hTNF sequence, into the AvaI-BstEII opened pPLc28BHTNF plasmid.
The plasmid contains the amp signal sequence, fused in frame to the mutated mature hTNF sequence.
The first 7 amino acids of the mature hTNFm28F are identical to wild type, but are encoded by the synthetic linker sequence.
Strains, containing pPLc28BHTNFm28F, secrete mature hTNFm28F into the periplasmic space, in a temperature dependent mode.
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.
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 R. Beyaert(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: Simon et al., Biotechnology 1 (1983), 784-791
Related plasmid reference: Van Ostade et al., FEBS Lett. 238 (1988), 347-352 [PMID: 3049162]
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 K514[pcI857]
Host reference: Zabeau et al., EMBO J. 1 (1982), 1217-1224 [PMID: 6327257]
Helper plasmid: pcI857
Cultivation medium: LB-Lennox + ampicillin (100 μg/ml) + kanamycin (50 μg/ml)
Cultivation temperature: 28°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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