Last data update: 24 January 2024 16:39 CET
Plasmid name: pMaLUC2 (LMBP 2837)
| New search | Print data sheet |
| 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: | - |
Refer in your Materials and Methods: |
pMaLUC2 (LMBP 2837) is available at BCCM/GeneCorner. This plasmid was deposited by Dr L. Steidler and Prof. Dr E. Remaut and was published in Steidler et al., 1996. |
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.