Last data update: 24 November 2020 04:18 CET
Plasmid name: pMcT4HTNFm29D37F (LMBP 2700)
|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 AA29 and AA37 of mature hTNF.
The mutagenesis of AA29 results in the loss of an AlwNI site, by which AlwNI became unique.
The mutagenesis of AA37 and the silent mutation of AA38 create a unique NruI site, and result in the loss of a XcmI, BalI, BglI, EaeI and BstXI site, by which the last 3 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.
|EMBL Accession number:||X01394.1, view at EMBL, GenBank, DDBJ|
|Latest sequence update:||16/12/1992|
|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.