Last data update: 24 November 2020 04:18 CET
Plasmid name: pKT240tet1 (LMBP 3162)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner core plasmid|
(View with Genome Compiler)
|Cloned DNA:||Broad-host-range plasmid RSF1010 plasmid mobilization genes A, B and C (mobA, mobB, mobC)
Broad-host-range plasmid RSF1010 plasmid replication genes A, B and C (repA, repB, repC)
Broad-host-range plasmid RSF1010 repressor F gene (rpsF)
|Selection marker:||Ampicillin (amp)
Neomycin (neo; kanamycin (kan))
|Replicon:||Broad-host-range plasmid RSF1010 vegetative replication origin; IncQ
Broad-host-range plasmid RSF1010 origin of transfer (oriT)
|Host range:||Escherichia coli
Gram-negative bacterial strains
|Parental clone:||pKT240; pBR322|
|Further information:||The plasmid was constructed by inserting the EcoRI-PvuII fragment from pBR322, containing the tetracycline resistance gene, between the EcoRI and NotI (filled-in with Klenow DNA polymerase) sites of pKT240. The structural streptomycin resistance genes (strA + strB) were lost due to the construction.
pKT240tet1 is a broad-host-range plasmid. It can be used in all gram-negative bacterial species that support replication of the natural plasmid RSF1010.
Introduce the plasmid by electroporation, or through conjugation, using plasmid RK2/RP4 transfer functions (provided in trans by RK2/RP4, RK2/RP4-derived vectors, other replicons carrying RK2/RP4 transfer functions (e.g. pRK2073) or strains expressing the tra genes of RK2/RP4).
The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727446.1.
The nucleotide sequence of the parental pKT240 is based on the information in Bagdasarian et al. (1983). The intention was to ligate the EcoRI-BstEII fragments of RSF1010 and pHSG415. However, the smallest EcoRI-BstEII fragment is approx. 1 kb bigger than expected. This region is indicated RSF' on the map as it contains NsiI and PstI sites, that fit with the RSF1010 sequence.
Since the nucleotide sequence of the plasmid could not be completely traced back, there is uncertainty as to the cutting frequency of the restriction enzymes, except for the sites that were analysed in the authenticity test.
The nucleotide sequence of RSF1010 corresponds with the EMBL Nucleotide Sequence Database accession number M28829.1.
Other name of the plasmid is pKT240Tet.
|EMBL Accession number:||M28829.1, view at EMBL, GenBank, DDBJ
LT727446.1, view at EMBL, GenBank, DDBJ
|Latest sequence update:||16/08/1995|
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: AvaI, EcoRI and SspI.|
|History of deposit:||This plasmid was deposited by Prof. Dr E. Remaut(1).
(1) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
|Related plasmid reference:||Scholz et al., Gene 75 (1989), 271-288 [PMID: 2653965]
|Restricted distribution:||- BCCM MTA|
|Distributed as:||H/P active culture or plasmid DNA|
|Host for distribution:||Escherichia coli K12 MC1061|
|Host reference:||Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493]
|Related host reference:||Brigé et al., Biochem. J. 394 (2006), 335-344 [PMID: 16293111]
|Cultivation medium:||LB-Lennox + ampicillin (100 μg/ml) + kanamycin (50 μg/ml) + tetracycline (10 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.