Last data update: 28 November 2020 04:24 CET
Plasmid name: pMaTrpMTNF (LMBP 2350)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner non-core plasmid|
|Cloned DNA:||Mouse tumor necrosis factor cDNA (Tnf, TNF); mature sequence|
|Promoter:||Escherichia coli tryptophan operon promoter (trp)|
|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 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.|
|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
|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)|
|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.