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LMBP plasmid details

Last data update: 28 November 2020 04:24 CET

Plasmid name: pMcTrpMTNFm29S32W (LMBP 2656)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p2656.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Mouse tumor necrosis factor cDNA (Tnf, TNF); L29S-R32W mutant
Promoter: Escherichia coli tryptophan operon promoter (trp)
Ribosome
binding site:
Ribosome binding site (RBS) of the Escherichia coli tryptophan operon attenuator peptide (trp)
Terminator: Phage fd terminator
Selection marker: Chloramphenicol (cam)
Replicon: Escherichia coli plasmid pMB1 origin
Phage f1 origin
Host range: Escherichia coli
Parental clone: pMaTrpMTNF
Further information: This plasmid was constructed by site-specific mutagenesis, where the nucleotide sequences of amino acids 29 and 32 of the mature mouse TNF (CTG, coding for Leu and CGC, coding for Arg) were converted to TCG (coding for Ser) and TGG (coding for Trp).
After mutagenesis, use mutS strains for primary transformation (e.g. sup(-) strain WK6mutS, sup(+) strain BMH71-18mutS); for segregation of possible mutants: sup(-) strains (e.g. WK6).
EMBL Accession number: M11731, view at EMBL, GenBank, DDBJ
Latest sequence update: 17/01/1993
Sequence detail:
RBS trp       | mature mouse TNF
                        | 1   2                               10
5'...    GTATCGATTCC.ATG.GTA.AGA.TCA.AGT.AGT.CAA.AAT.TCG.AGT.GAC.AAG.CCT.GTA.GCC.CAC.GTC.GTA.
                  NcoI   * *         **  **          TaqI
                         Val 

          20                                  29          32                       
.GCA.AAC.CAC.CAA.GTG.GAG.GAG.CAG.CTG.GAG.TGG.TCG.AGC.CAG.TGG.GCC.AAC ...3'
                                             **          **
                                             TaqI         HaeIII 
                                             Ser         Trp
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).
The plasmid was constructed by Dr Xaveer Van Ostade.
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: Van Ostade et al., Eur. J. Biochem. 220 (1994), 771-779 [PMID: 8143731]
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 SURE
Host reference: Greener, Strategies 3 (1990), 5-6
Cultivation medium: LB-Lennox + chloramphenicol (25 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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