GREAT AT SMALL THINGS

0

LMBP plasmid details

Last data update: 24 November 2020 04:18 CET

Plasmid name: pKT240tet2 (LMBP 3226)

Add to cart

New search

Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p3226.gb (View with Genome Compiler)
p3226.txt
p3226.pdf
Sequence
analysis results
Genecorner:

-

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)
Promoter: -
Ribosome
binding site:
-
Terminator: -
Selection marker: Tetracycline (tet)
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: pKT240tet1
Further information: The plasmid was constructed by cutting, blunting (Klenow DNA polymerase) and ligating the PstI and XhoI sites of pKT240tet1. The structural streptomycin resistance genes (strA + strB), the ampicillin and kanamycin resistance genes of pKT240 were lost.
pKT240tet2 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 LT727452.1.
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. The PstI site inside this fragment is used for the construction of this plasmid. The remnant region is indicated RSF' on the map.
Since the nucleotide sequence of the pKT240tet2 vector 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 pKT240TetdKan.
EMBL Accession number: M28829.1, view at EMBL, GenBank, DDBJ
LT727452.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 16/08/1995
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BamHI, BstXI, EcoRV and NcoI.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Prof. Dr E. Remaut(1).
(1) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: -
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 + tetracycline (10 μg/ml)
Cultivation temperature: 37°C
Biosafety level: L1
Cultivation remark: -
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