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GeneCorner plasmid details

Last data update: 02 December 2020 04:29 CET

Plasmid name: pBLhIL6LUC (LMBP 3287)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p3287.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Photinus pyralis (firefly) luciferase gene (LUC)
Promoter: Human interleukin 6 promoter (IL6)
Escherichia coli lac operon promoter
Ribosome
binding site:
-
Terminator: Simian virus 40 polyadenylation signal (SV40 polyA)
Selection marker: Ampicillin (amp)
Replicon: Escherichia coli plasmid pMB1 origin
Host range: Escherichia coli; preferably recombination-deficient strains
Mammalian cells
Parental clone: pSV2L5; pBLHIL6CAT
Further information: The plasmid was constructed as follows: the luciferase gene of pSV2L5 was inserted as a BamHI-HindIII fragment between the XhoI and XmaI (SmaI) sites of pBLHIL6CAT, replacing the chloramphenicol acetyltransferase gene. The restriction sites involved were filled in with Klenow DNA polymerase, restoring the HindIII, BamHI and XmaI (SmaI) sites.
The resulting plasmid allows functional analysis of the human IL6 promoter.
This plasmid contains the SV40 small t-antigen splicing signal. It also contains three SV40 polyadenylation segments. Hence, the possibility might exist that the plasmid could loose information by homologous recombination.
When cloning a fragment downstream from the lac promoter it may be advisable to use lacI(q) strains in order to prevent fortuitous expression of a possibly noxious polypeptide.
The nucleotide sequence of the genomic hIL6 DNA corresponds with the EMBL Nucleotide Sequence Database accession number AC073072.11. However, the C nucleotide at promoter position -174 (polymorphism mutant) was changed into a G nucleotide (wild type).
Other name of the plasmid is 1168Luc.
EMBL Accession number: AC073072.11, view at EMBL, GenBank, DDBJ
Latest sequence update: 30/04/2004
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: AvaII, BamHI, HindIII/PvuII, NheI/XmaI, SalI and StyI/XbaI.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Prof. Dr R. Beyaert(1) (2). It was constructed in the research unit of Em. Prof. Dr G. Haegeman(2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: -
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)
Cultivation temperature: 37°C
Biosafety level: L1
Cultivation remark: -
Other culture collection numbers: -

Refer in your Materials and Methods:

pBLhIL6LUC (LMBP 3287) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr R. Beyaert .

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.

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