Last data update: 24 November 2020 04:18 CET
Plasmid name: pPLGN1 (LMBP 1179)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner non-core plasmid|
|Cloned DNA:||Aminoglycoside phosphotransferase structural gene (aph); strA+strB genes (conferring streptomycin resistance) without promoter
Phage λ cI temperature-sensitive repressor gene (cIts, cI857)
|Promoter:||Phage λ major leftward promoter (λ PL)
Phage λ major rightward promoter (λ PR)
Phage λ promoter for repressor maintenance (λ PRM)
|Selection marker:||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
|Further information:||The plasmid was constructed by inserting the cI gene (cI857 mutant) from phage λ cI857 Sam7, coding for the λ repressor protein, as a BglII-PstI fragment into the unique BamHI and the PstI (nucleotide position 7774) sites of pPL1010, replacing the ampicillin resistance gene.
pPLGN1 is a broad-host-range plasmid. It can be used in all gram-negative bacterial species that support replication of the natural plasmid RSF1010.
Useful vector for λ PL-regulated expression of cloned genes after heat inactivation of the cIts function.
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). Propagation temperatures may not exceed 34°C, preferentially grow at 28°C.
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 smalest 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.
The nucleotide sequence of RSF1010 corresponds with the EMBL Nucleotide Sequence Database accession number M28829.1.
Other name of the plasmid is pPLCI1010.
|EMBL Accession number:||M28829.1, view at EMBL, GenBank, DDBJ|
|Latest sequence update:||25/03/2011|
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: AvaI, BstXI, EcoRI and SspI.|
|History of deposit:||This plasmid was deposited by R. Leemans 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 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 + kanamycin (50 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.