Last data update: 28 November 2020 04:24 CET
Plasmid name: pMc59TACBLAHI (LMBP 2269)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner non-core plasmid|
|Cloned DNA:||Ampicillin resistance gene (amp)
Human immunoglobulin γ3 cDNA (hIG3); heavy chain hinge region
|Promoter:||Escherichia coli hybrid tryptophan/lacUV5 promoter (tac)|
|Terminator:||Phage fd terminator|
|Selection marker:||Ampicillin (amp)
|Replicon:||Escherichia coli plasmid pMB1 origin
Phage f1 origin
|Host range:||Escherichia coli|
|Parental clone:||pMc59BLAHI; pMc519|
|Further information:||The plasmid was constructed as follows : the PvuII-BamHI fragment of pMc59BLAHI was substituted by the PvuII-BamHI (nucleotide position 3857) fragment of pMc519, resulting in the insertion of the tac promoter before the BLA-HI fusion gene
This pMc59BLAHI-derivative contains the tac promoter upstream to the BlaHi fusion gene.
This plasmid provides resistance to chloramphenicol. It also provides resistance to ampicillin even without induction of the tac promoter. This resistance to ampicillin could partly be due to leakage of the tac promoter, but according to the features of the parental clone pMc59BLAHI, we may also conclude that the constitutive promoter of kanamycin phosphoribosyltransferase is still present in a functional way right in front of the fusion gene.
The translation of the fusion gene stops 8 amino acids behind the end of the hinge region of hIG3 pMc59TACBLAHI contains the replication origin of the single-stranded DNA phage F1, so that it can be rapidly switched between the plasmid and the phage mode (ssDNA) of replication.
When cloning a fragment downstream from the tac promoter it may be advisable to use lacI(q) strains in order to prevent fortuitous expression of a possibly noxious polypeptide.
Other name of the plasmid is pMc59TACBLAHI1.
|EMBL Accession number:||-|
|Latest sequence update:||23/01/1997|
|Authenticity test:||The plasmid still needs to be subjected to the authenticity test.|
|History of deposit:||This plasmid was deposited by Dr K. De Sutter(1).
(1) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
|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) + chloramphenicol (25 μg/ml)*|
|Cultivation remark:||*: selection of transformants on chloramphenicol; subsequent cultivation of a single colony in liquid medium with ampicillin.|
|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.