Last data update: 15 January 2021 04:14 CET
Plasmid name: p1168hIL6mCRE-luc+ (LMBP 4499)
|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)
Synthetic polyadenylation signal (polyA)
|Selection marker:||Ampicillin (amp)|
|Replicon:||Escherichia coli plasmid pMB1 origin
Phage f1 origin
|Host range:||Escherichia coli
|Parental clone:||pGL3-Basic; pMahIL6P|
|Further information:||The plasmid was constructed by inserting the XmaI/HindIII fragment of pMahIL6P, containing the human IL6 promoter, between the XmaI and HindIII sites of pGL3-Basic.
The binding site for the cyclic AMP-responsive element-binding protein (CRE) transcription factor of the human IL6 promoter was abolished by site-directed mutagenesis, creating an additional BamHI site.
The SalI site between the SV40 polyA and the pMB1 origin was filled in. However, the resulting PvuI site could not be experimentally confirmed.
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 LT727234.1.
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).
Other name of the plasmid is p1168huIL6Pluc+CRE(BamHI)mut*.
|EMBL Accession number:||AC073072.11, view at EMBL, GenBank, DDBJ
LT727234.1, view at EMBL, GenBank, DDBJ
|Latest sequence update:||29/04/2004|
Mutator oligonucleotide used: CRE mutant 5' GCGATGCTAAAGGGATCCACATTGCACAAT 3' BamHI
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: BamHI, BglI/SphI, HindIII, NcoI/SalI, PvuI and SphI.|
|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]
Bogard et al., Naunyn Schmiedebergs Arch. Pharmacol. 387 (2014), 329-339 [PMID: 24363043] [DOI: 10.1007/s00210-013-0950-4]
|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.