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LMBP plasmid details

Last data update: 28 November 2020 04:24 CET

Plasmid name: pMaTrpMTNF (LMBP 2350)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p2350.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Mouse tumor necrosis factor cDNA (Tnf, TNF); mature sequence
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: Ampicillin (amp)
Replicon: Escherichia coli plasmid pMB1 origin
Phage f1 origin
Host range: Escherichia coli
Parental clone: pMa58; pATTrpMTNF
Further information: This plasmid was constructed by inserting the PstI-XbaI fragment of pATTrpMTNF (containing mTnf and the trp promotor into unique PstI and unique XbaI sites of pMa58. The unique SalI site of the polylinker of pMa58 has been deleted. The PstI restriction site of the ampicillin resistance gene has been removed, but the plasmid conserves its AMP resistance capacity, and in a Sup- host strain it is sensitive to CAM.
The exact sequence between the PstI site and the lost PvuII site of the trp promoter sequence is unknown.
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: 20/12/1991
Sequence detail:
Sequence at the start of mature mTnf:

                      signal sequence| mature mouse Tnf
mTNF wild type: 5' ...CAG.ACC.CTC.ACA.CTC.AGA.TCA.TCT.TCT.CAA.AAT.TCG.AGT…3'
                      Gln Thr Leu Thr Leu Arg Ser Ser Ser Gln Asn Ser Ser

                       RBS trp       | mature mouse Tnf
pMaTrpMTNF:     5'    GTATCGATTCC.ATG.GTA.AGA.TCA.AGT.AGT.CAA.AAT.TCG.AGT…3'
                                  Met Val Arg Ser Ser Ser Gln Asn Ser Ser
                               NcoI
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: -
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 DH1(λ)
Host reference: Bachmann, in 'Escherichia coli and Salmonella: Cellular and Molecular Biology', American Society for Microbiology, Washington DC (1987), 1190-1219
Cultivation medium: LB-Lennox + ampicillin (100 μg/ml)
Cultivation temperature: 37°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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