Last data update: 28 November 2020 04:24 CET
Plasmid name: pMc58SA (LMBP 4329)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner non-core plasmid|
|Cloned DNA:||Streptomyces avidinii streptavidin gene (SA)|
|Terminator:||Phage fd terminator|
|Selection marker:||Chloramphenicol (cam)|
|Replicon:||Escherichia coli plasmid pMB1 origin
Phage f1 origin
|Host range:||Escherichia coli|
|Parental clone:||pMc58; pSA307|
|Further information:||The plasmid was constructed by cloning the EcoRI-HindIII fragment from pSA307, containing the Streptomyces avidinii streptavidin gene, between the unique EcoRI and HindIII sites of pMc58.
The sequence of the streptavidin gene was obtained from 'GENBANK' (Release 55; Locus: STMSTRAVG) and shows as compared to Dr Lieberman's sequence map of the parental clone pSA307, a few differences in the 5' and 3' untranslated regions of the gene.
This phasmid provides resistance to chloramphenicol but is sensitive to ampicillin.
More than one mutagenesis round is possible by switching from chloramphenicol resistance to ampicillin resistance.
pMc58SA 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 a helper phage M13KO7.
After mutagenesis, use mutS strains for primary transformation (e.g. sup(-) strain WK6mutS, sup(+) strain BMH71-18mutS); for segregation of possible mutants: sup(-) strains (e.g. WK6).
|EMBL Accession number:||-|
|Latest sequence update:||10/07/1990|
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: BamHI, BglI and EcoRI/HindIII. The plasmid seems to be approx. 200 bp larger as compared to the compiled nucleotide sequence, the extra nucleotides being located in the vector part of the plasmid.|
|History of deposit:||This plasmid was deposited by J. Viaene(1) and Prof. Dr E. Remaut(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 + chloramphenicol (25 μg/ml)|
|Other culture collection numbers:||LMBP 01992|
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.