Last data update: 28 November 2020 04:24 CET
Plasmid name: pMahTNFR55fm (LMBP 2784)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner core plasmid|
(View with Genome Compiler)
|Cloned DNA:||Human tumor necrosis factor receptor p55 cDNA (TNFR55, TNFR1, TNFRSF1A); part of the mutated mature sequence|
|Terminator:||Phage fd terminator|
|Selection marker:||Ampicillin (amp)|
|Replicon:||Escherichia coli plasmid pMB1 origin
Phage f1 origin
|Host range:||Escherichia coli|
|Parental clone:||pMa58; pUChTNFR55|
|Further information:||This plasmid was constructed as follows: the PvuII fragment from pUChTNFR55, containing part of the mature sequence of hTNFR55, was inserted into the unique SmaI site of pMa58.
The hTNFR55 fragment contains part of the extracellular region encoding amino acids 73 up to and including 182, the transmembrane domain encoding amino acids 183 up to and including 203, the intracellular region encoding amino acids 204 up to and including 427 and part of the 3' UTR.
Following site-specific mutagenesis, an ATG initiation codon was introduced at the end of the transmembrane domain, encoding amino acid 201 of the mature hTNFR55. At the same time, a unique NcoI site was introduced.
The start position of the intracellular part of hTNFR55 differs from one publication to another. Here the description of Dr Loetscher is used.
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 LT727543.1.
Other name of the plasmid is pMaHTNFR1fm.
|EMBL Accession number:||LT727543.1, view at EMBL, GenBank, DDBJ|
|Latest sequence update:||16/11/1994|
Nucleotide sequence of the mutated hTNFR55 cDNA: transmembrane domain -->| | 200 |204 hTNFR55wt: 5' TTA TCC CTC CTC TTC ATT GGT TTA ATG TAT CGC 3' Leu Ser Leu Leu Phe Ile Gly Leu Met Tyr Arg | 200 |204 hTNFR55m: 5' TTA TCC CTC CTC TCC ATG GGT TTA ATG TAT CGC 3' Leu Ser Leu Leu Ser Met Gly Leu Met Tyr Arg NcoI linker 3' GG GAG GAG AGG TAC CCA AAT TAC ATA GCG 5'
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: EcoRI and NcoI.|
|History of deposit:||This plasmid was deposited by Prof. Dr G. Haegeman(1). It was constructed by Dr V. Vandevoorde(1).
(1) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
|Plasmid reference:||Vandevoorde et al., J. Cell Biol. 137 (1997), 1627-1638 [PMID: 9199176]
|Restricted distribution:||- BCCM MTA|
|Distributed as:||H/P active culture or plasmid DNA|
|Host for distribution:||Escherichia coli K12 MC1061|
|Host reference:||Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493]
|Related host reference:||Brigé et al., Biochem. J. 394 (2006), 335-344 [PMID: 16293111]
|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.