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LMBP plasmid details

Last data update: 24 November 2020 04:18 CET

Plasmid name: pMaLUC1 (LMBP 2810)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: pmaluc1.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: pMa58; pSV2L5
Further information: The plasmid was constructed by inserting a 5' fragment of the LUC gene (EcoRI- BamHI fragment of pSV2L5) between the EcoRI and BamHI sites of pMa58.
The plasmid contains a fragment (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 was designed for site-specific mutagenesis where the mutated sequence can be physically linked with a selectable marker (cam).
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: 04/04/1995
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: LMBP 02813

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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