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GeneCorner plasmid details

Last data update: 05 December 2020 04:15 CET

Plasmid name: pBLHIL6CAT (LMBP 2861)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p2861.gb (View with Genome Compiler)
p2861.pdf
Sequence
analysis results
Genecorner:

-

Cloned DNA: Chloramphenicol acetyltransferase gene (cat)
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: pBLCAT5; pGEM1gHIL61
Further information: The plasmid was constructed by replacing the BamHI-XhoI fragment, containing the HSV-TK promoter, of pBLCAT5 by the 1186 bp BamHI-XhoI fragment, containing the hIL6 promoter, of pGEM1gHIL61.
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 polyA segments. Hence, the possibility might exist that the plasmid could loose information by homologous recombination. So far, no observations have been made in this respect.
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 this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727549.1.
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 names of the plasmid are pBLCAT5HIL6 and pBLHIL6CAT5.
EMBL Accession number: AC073072.11, view at EMBL, GenBank, DDBJ
LT727549.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 29/04/2004
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BamHI/XhoI, HindIII, NheI, SphI, SspI and StyI.
Class: Recombinant plasmid
Type: Plasmid
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: -
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 DH5α
Host reference: Focus 8 (1986), 9
Related host reference: Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051]
Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187]
Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791]
Rodriguez-Quinones et al., Focus 15 (1993), 110-112
Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660]
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:

pBLHIL6CAT (LMBP 2861) is available at BCCM/GeneCorner. This plasmid was deposited by Dr W. Vanden Berghe and Prof. Dr G. Haegeman.

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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