Last data update: 29 October 2020 04:15 CET
Plasmid name: p1168hIL6m3AP1-luc+ (LMBP 4492)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner core plasmid|
(View with Genome Compiler)
|Cloned DNA:||Photinus pyralis (firefly) luciferase gene (LUC); mutated coding region (LUCm; luc(+))|
|Promoter:||Human interleukin 6 promoter (IL6); mutated sequence|
|Terminator:||Simian virus 40 polyadenylation signal (SV40 polyA)|
|Selection marker:||Ampicillin (amp)|
|Replicon:||Escherichia coli plasmid pMB1 origin|
|Host range:||Escherichia coli
|Parental clone:||pGL3-Basic; pBLHIL6CAT|
|Further information:||The plasmid was constructed by ligating the 2338 bp AseI (VspI)/XhoI fragment of pBLHIL6CAT, containing the human IL6 promoter, to the 3293 bp AseI (VspI)/XhoI fragment of pGL3-Basic.
The 3' binding site for the activator protein 1 (AP1) transcription factor of the human IL6 promoter was abolished by site-directed mutagenesis, creating a unique EcoRI site.
Mammalian cells can be directly transfected transiently with the plasmid. However, promoter regulation can be better evaluated when the mammalian cells are stably transfected.
The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727228.1.
The nucleotide sequence of the genomic human IL6 DNA corresponds with the EMBL Nucleotide Sequence Database accession number AC073072.11, except for the C nucleotide at promoter position -174 (polymorphism mutant) which was changed into a G nucleotide (wild type) and for the abolished 3AP1 site.
Other name of the plasmid is p1168hu.IL6P-luc+3AP1(EcoRI)mut.
|EMBL Accession number:||AC073072.11, view at EMBL, GenBank, DDBJ
LT727228.1, view at EMBL, GenBank, DDBJ
|Latest sequence update:||30/04/2004|
Mutator oligonucleotide used: 3'AP1 mutant 5' GGGATTTTCCCAGAATTCTCAATATTAG 3' EcoRI
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: Eco47III/XbaI, EcoRI, NcoI/SalI and XhoI.|
|History of deposit:||This plasmid was deposited by Dr W. Vanden Berghe(1) and Prof. Dr G. Haegeman(1).
(1) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
|Plasmid reference:||Vanden Berghe et al., J. Biol. Chem. 273 (1998), 3285-3290 [PMID: 9452444]
|Related plasmid reference:||Vanden Berghe et al., J. Biol. Chem. 274 (1999), 32091-32098 [PMID: 10542243]
|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.