Last data update: 28 November 2020 04:24 CET
Plasmid name: pMc59BLAHI (LMBP 2268)
|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
|Terminator:||Phage fd terminator|
|Selection marker:||Ampicillin (amp)
|Replicon:||Escherichia coli plasmid pMB1 origin
Phage f1 origin
|Host range:||Escherichia coli|
|Parental clone:||pMc59; pMac254BLAHI|
|Further information:||The plasmid was constructed by ligating the BssHII (nucleotide position 2926; filled in with Klenow DNA polymerase) - AlwNI (blunted with T4 DNA polymerase) fragment of pMac254BLAHI, containing the fusion gene, in clockwise orientation in the unique XbaI site (filled in with Klenow DNA polymerase) of pMc59.
This pMc59-derivative is similar to pMac254BLAHI, but the fusion gene is clockwise orientated and located between the chloramphenicol resistance and the fd terminators.
The phasmid provides resistance to chloramphenicol. Since it provides also resistance to ampicillin, one can conclude that the constitutive promoter of kanamycin phosphoribosyltransferase is still present in a functional way upstream of the fusion gene.
The translation of the fusion gene stops 8 amino acids behind the end of the hinge region of the human IgG3 heavy chain.
pMc59BLAHI 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. The latter requires infection with the helper phage M13KO7.
|EMBL Accession number:||-|
|Latest sequence update:||22/01/1997|
|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.