GREAT AT SMALL THINGS

0

LMBP plasmid details

Last data update: 28 November 2020 04:24 CET

Plasmid name: pMc59TACBLAHI (LMBP 2269)

Add to cart

New search

Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p2269.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Ampicillin resistance gene (amp)
Human immunoglobulin γ3 cDNA (hIG3); heavy chain hinge region
Promoter: Escherichia coli hybrid tryptophan/lacUV5 promoter (tac)
Ribosome
binding site:
-
Terminator: Phage fd terminator
Selection marker: Ampicillin (amp)
Chloramphenicol (cam)
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.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Dr K. De Sutter(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) + chloramphenicol (25 μg/ml)*
Cultivation temperature: 37°C
Biosafety level: L1
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.

New search