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GeneCorner plasmid details

Last data update: 20 October 2020 04:14 CEST

Plasmid name: pATTrpMTNFm29S32W (LMBP 2658)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p2658.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: -
Selection marker: Tetracycline (tet)
Replicon: Escherichia coli plasmid pMB1 origin
Host range: Escherichia coli; use strains expressing the trpR gene coding for the tryptophan operon repressor protein (e.g. K514).
Parental clone: pATTrpMTNF; pMcTrpMTNFm29S32W
Further information: This plasmid was constructed as follows : the EcoRI-PstI fragment of pMcTrpMTNFm29S32W (containing mutant MTNF)
was inserted into the unique EcoRI and PstI restriction sites of pATTrpMTNF, replacing the MTNF wild type.
The first 8 amino acids of mature MTNF are encoded by a synthetic NcoI-TaqI linker, that only differs for the first amino acid, where Leu is replaced by Val.
In the mouse TNF L29S-R32W mutant the nucleotide sequences of amino acids 29 and 32 of the wild type mature mouse TNF (CTG, coding for Leu and CGC, coding for Arg) are converted to TCG (coding for Ser) and TGG (coding for Trp).
EMBL Accession number: M11731, view at EMBL, GenBank, DDBJ
Latest sequence update: 28/01/1993
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). It was constructed by Dr X. 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 + tetracycline (10 μg/ml)
Cultivation temperature: 37°C
Biosafety level: L1
Cultivation remark: -
Other culture collection numbers: -

Refer in your Materials and Methods:

pATTrpMTNFm29S32W (LMBP 2658) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr R. Beyaert and was published in Van Ostade et al., 1994.

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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