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LMBP plasmid details

Last data update: 24 November 2020 04:18 CET

Plasmid name: pPL1010 (LMBP 1121)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p1121.gb (View with Genome Compiler)
p1121.txt
p1121.pdf
Sequence
analysis results
Genecorner:

-

Cloned DNA: Aminoglycoside phosphotransferase structural gene (aph); strA+strB genes (conferring streptomycin resistance)
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: Phage λ major leftward promoter (λ PL)
Ribosome
binding site:
-
Terminator: -
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; use strans with a cI function, cIts for PL controlled expression
Parental clone: pKT240; pPLa832
Further information: The plasmid was constructed by excising the EcoRI-HindIII fragment from pKT240 and replacing it by the EcoRI-HindIII fragment from pPLa832, restoring the kanamycin resistance gene and introducing the λ PL promoter.
pPL1010 contains the structural streptomycin resistance genes (strA + strB) under control of the λ PL promoter.
pPL1010 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 LT727568.1.
The nucleotide sequence of this plasmid is based on the information for pKT240 in Bagdasarian et al. (1983). The intention was to ligate 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 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
LT727568.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 27/03/1995
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BstXI, EcoRI, HincII, NcoI, NsiI/ScaI and SspI.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by R. Leemans(1) and Prof. Dr E. Remaut(1).
(1) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: Leemans et al., J. Bacteriol. 169 (1987), 1899-1904 [PMID: 2952635]
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 ΔH1Δtrp
Host reference: Bernard et al., Gene 5 (1979), 59-76 [PMID: 372049]
Related host reference: Remaut et al., Gene 15 (1981), 81-93 [PMID: 6271633]
Cultivation medium: LB-Lennox + ampicillin (100 μg/ml) + kanamycin (50 μg/ml)
Cultivation temperature: 28°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.

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