Last data update: 28 November 2020 04:24 CET
Plasmid name: pMc254 (LMBP 2098)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner core plasmid|
(View with Genome Compiler)
|Terminator:||Phage fd terminator|
|Selection marker:||Chloramphenicol (cam)|
|Replicon:||Escherichia coli plasmid pMB1 origin
Phage f1 origin
|Host range:||Escherichia coli|
|Further information:||The plasmid was designed for site-specific mutagenesis where the mutated sequence can be physically linked with a selectable marker (ampicillin).
The plasmid provides resistance to chloramphenicol, but is sensitive to ampicillin.
More than one mutagenesis round is possible via switching from chloramphenicol resistance to ampicillin resistance.
pMc254 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, e.g. M13KO7.
The f1 part of the plasmid contains the f1 origin but also the sequence around the gene II promoter.
The two trp terminators at the end of the chloramphenicol gene are not present.
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).
The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727619.1.
|EMBL Accession number:||LT727619.1, view at EMBL, GenBank, DDBJ|
|Latest sequence update:||10/11/1989|
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: BsrDI, MspA1I and PdiI.|
|History of deposit:||This plasmid was deposited by Dr P. Stanssens(1).
(1) Corvas International N.V., Ghent, Belgium
|Restricted distribution:||- BCCM MTA|
|Distributed as:||H/P active culture or plasmid DNA|
|Host for distribution:||Escherichia coli K12 F165|
|Cultivation medium:||LB-Lennox + chloramphenicol (25 μg/ml)|
|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.