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LMBP plasmid details

Last data update: 24 November 2020 04:18 CET

Plasmid name: pKT212 (LMBP 8021)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: -
Sequence
analysis results
Genecorner:

-

Cloned DNA: Broad-host-range plasmid RSF1010 plasmid replication genes A, B and C (repA, repB, repC)
Broad-host-range plasmid RSF1010 repressor F gene (rpsF)
Aminoglycoside phosphotransferase structural gene (aph); strA and strB genes (conferring streptomycin resistance) without promoter
Broad-host-range plasmid RSF1010 plasmid mobilization genes A, B and C (mobA, mobB, mobC)
Promoter: -
Ribosome
binding site:
-
Terminator: -
Selection marker: Chloramphenicol (cam)
Streptomycin (Sm)
Tetracycline (tet)
Replicon: Broad-host-range plasmid RSF1010 origin of transfer (oriT)
Broad-host-range plasmid RSF1010 vegetative replication origin; IncQ
Host range: Escherichia coli
Gram-negative bacterial strains
Parental clone: pKT210; pKT007
Further information: The plasmid was created by ligating a HindIII fragment of pKT007 into the HindIII opened pK210 vector. pKT007 was obtained by transposing the Tn10 fragment of R6 onto pML21.
pKT212 is a broad-host-range plasmid. It can be used in all gram-negative bacterial species that support replication of the natural plasmid RSF1010.
The plasmid contains the structural streptomycin resistance genes (strA + strB), lacking a functional promoter, and can be used as a promoter probe vector.
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 is based on the information in Bagdasarian et al. (1983).
Since the nucleotide sequence 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.
EMBL Accession number: M28829.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 18/03/2013
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: EcoRV, HindIII, NcoI and NotI.

As compared to the compiled sequence file, the expected 6445 bp NcoI fragment has a size of around 5300.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by M. Mergeay(1) and A. Provoost(1).
(1) SCK.CEN, Mol, Belgium
Plasmid reference: -
Related plasmid reference: Bagdasarian et al., in 'Plasmids of Medical, Environmental and Commercial Importance', Elsevier/North Holland Biomedical Press (1979), 411-422 [ISSN/ISBN: 0444801618]
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 SK1592
Host reference: -
Cultivation medium: LB-Lennox + chloramphenicol (34 μg/ml) + streptomycin (25 μg/ml) + tetracycline (10 μg/ml)*
Cultivation temperature: 37°C
Biosafety level: L1
Cultivation remark: *: selection of transformants on chloramphenicol; subsequent cultivation of a single colony in liquid medium with streptomycin and tetracycline.
Other culture collection numbers: SCK CM0262

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.

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