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LMBP plasmid details

Last data update: 28 November 2020 04:24 CET

Plasmid name: pMaLUC2 (LMBP 2837)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: pmaluc2.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Photinus pyralis (firefly) luciferase gene (LUC); 5' fragment (198 AA of 550)
Promoter: -
Ribosome
binding site:
-
Terminator: Phage fd terminator
Selection marker: Ampicillin (amp)
Replicon: Escherichia coli plasmid pMB1 origin
Phage f1 origin
Host range: Escherichia coli
Parental clone: pMaLUC1
Further information: The plasmid was constructed by in vitro mutagenesis of pMaLUC1, creating unique restriction sites at the start of the LUC gene.
The plasmid contains a fragment (the first 198 aa of 550) of the intronless hybrid genomic-cDNA luciferase gene from the firefly Photinus pyralis, missing 17 nucleotides of the 5' flanking DNA.
The phasmid provides resistance to ampicillin, but is sensitive to chloramphenicol.
More than one mutagenesis round is possible via switching from ampicillin resistance to chloramphenicol resistance.
pMa58 contains the replication origin of the single-stranded DNA phage f1 (clockwise orientated), so that it can be rapidly switched between the plasmid and the phage mode (ssDNA) of replication. The latter requires infection with a helper phage, e.g. M13KO7.
After mutagenesis, use mutS strains for primary transformation (e.g. sup(-) strain WK6mutS, sup(+) strain BMH71-18mutS); for segregation of possible mutants: sup(-) strains (e.g. WK6).
EMBL Accession number: -
Latest sequence update: 05/04/1995
Sequence detail:
wild type LUC gene :  GTTGGTAAA ATG GAA GAC

mutant             :  GTTGGTACC ATG GAA GAC
                             **
                             NcoI
                         KpnI

* : mutated nucleotide
Authenticity test: The plasmid still needs to be subjected to the authenticity test.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Dr L. Steidler(1) (2) and Prof. Dr E. Remaut(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: Steidler et al., Appl. Environ. Microbiol. 62 (1996), 2356-2359 [PMID: 8779573]
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 + chloramphenicol (25 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.

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