Last data update: 28 November 2020 04:24 CET
Plasmid name: pMcT4HTNFm38V (LMBP 2630)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner non-core plasmid|
|Cloned DNA:||Human tumor necrosis factor cDNA (TNF); mutated mature sequence|
|Promoter:||Phage T4 gene 32 promoter (T4g32)|
|Ribosome binding site (RBS) of the phage T4 gene 32 (T4g32)|
|Terminator:||Phage fd terminator
Escherichia coli tryptophan operon terminator (trp); synthetic sequence
|Selection marker:||Chloramphenicol (cam)|
|Replicon:||Escherichia coli plasmid pMB1 origin
Phage f1 origin
|Host range:||Escherichia coli|
|Further information:||The plasmid was constructed by site-specific mutagenesis of pMa-T4-hTNF-TA. As a result, codon 38 (GCC) of mature hTNF encoding alanine (A) was replaced by GTC encoding valine (V), resulting in the loss of the unique BalI site and in the loss of a BstXI, EaeI and BglI site, by which these sites became unique in the plasmid.
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).
The nucleotide sequence of the wt hTNF cDNA corresponds with the EMBL Nucleotide Sequence Database accession number X01394.1.
Other name of the plasmid is pMcT4HTNFTm38V.
|EMBL Accession number:||X01394.1, view at EMBL, GenBank, DDBJ|
|Latest sequence update:||04/01/1993|
|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 SURE|
|Host reference:||Greener, Strategies 3 (1990), 5-6
|Cultivation medium:||LB-Lennox + chloramphenicol (25 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.