Last data update: 24 November 2020 04:18 CET
Plasmid name: pMa-T4-hTNF-TA (LMBP 2345)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner core plasmid|
(View with Genome Compiler)
|Cloned DNA:||Human tumor necrosis factor cDNA (TNF); 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:||Ampicillin (amp)|
|Replicon:||Escherichia coli plasmid pMB1 origin
Phage f1 origin
|Host range:||Escherichia coli|
|Parental clone:||pMa58; pPLT4HTNFST8|
|Further information:||This plasmid was constructed by inserting the EcoRI fragment from pPLT4HTNFST8, containing the phage T4 promoter, the hTNF mature sequence and the synthetic trpA terminator, clockwise into the unique EcoRI site of pMa58.
The unique SalI site of the polylinker of pMa58 was deleted.
Although the PstI restriction site of the ampicillin resistance gene was removed, the plasmid conserves its resistance to ampicillin, but it is sensitive to chloramphenicol (in sup(-) host strains).
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 this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727505.1.
The nucleotide sequence of the hTNF cDNA corresponds with the EMBL Nucleotide Sequence Database accession number X01394.1.
Other name of the plasmid is pMaT4hTNFT.
|EMBL Accession number:||X01394.1, view at EMBL, GenBank, DDBJ
LT727505.1, view at EMBL, GenBank, DDBJ
|Latest sequence update:||04/02/1992|
Nucleotide sequence at the 5' end of the mature human TNF cDNA: T4 promoter + RBS -->| |--> mature human TNF 5' ... AAATTAAAAAGGAAAT|CGATACTACT.ATG.GTC.AGA.TCA.TCT.TCT.CGA.ACC.CCG. ... 3' ClaI Met Val Arg ser Ser Ser Arg Thr Pro
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: AvaI, BstEII, EcoRI and PstI.|
|History of deposit:||This plasmid was deposited by Prof. Dr F. Van Roy(1) (2). It was constructed by X. Van Ostade(1) (2).
(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., FEBS Lett. 238 (1988), 347-352 [PMID: 3049162]
|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.