Last data update: 24 January 2024 16:39 CET
Plasmid name: pJBtrcLUC (LMBP 3134)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p3134.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Photinus pyralis (firefly) luciferase gene (LUC) Escherichia coli lac repressor gene (lacI) Broad-host-range plasmid RK2 trans-acting replication initiator protein DNA (trfA) |
| Promoter: | Transposon Tn5 neomycin resistance gene promoter (neo) Escherichia coli lac repressor promoter; mutant (lacI(q)) Escherichia coli hybrid tryptophan/lacUV5 promoter (trc) |
| Ribosome binding site: |
Ribosome binding site (RBS) of the Escherichia coli lac Z gene (lacZ) |
| Terminator: | - |
| Selection marker: | Tetracycline (tet) |
| Replicon: | Broad-host-range plasmid RK2 vegetative replication origin Broad-host-range plasmid RK2/RP4 origin of transfer (oriT) |
| Host range: | Escherichia coli Gram-negative bacterial strains |
| Parental clone: | pJBtrc; pMaLUC2; pSV2L5 |
| Further information: | The plasmid was constructed as follows: first the EcoRI-NcoI fragment of pMaLUC2 (containing the 5' part of the luciferase gene) was inserted between the EcoRI and NcoI sites of the polylinker of pJBtrc. Then the EcoRI fragment of pSV2L5 (containing the 3' part of the luciferase gene) was inserted into the EcoRI site of the polylinker of the intermediate construct, restoring the gene. The plasmid contains the full-length and intronless luciferase gene from the firefly Photinus pyralis, derived from genomic DNA (from -30 through +68) and cDNA (from +69 through +1480). 17 nucleotides of the 5' flanking DNA and 30 nucleotides of the 5' untranslated region are missing. The luciferase gene can be used as an efficient reporter gene. This IncP plasmid is used as a broad-host-range vector. The plasmid RK2 trfA gene encodes two polypeptide products P382 and P285 of 382 and 285 amino acids respectively, using a different initiation codon within the same reading frame. The resulting protein is essential for vegetative replication. 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). |
| EMBL Accession number: | - |
| Latest sequence update: | 18/04/1995 |
| 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 K. Keymeulen(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: | Remaut et al., in 'Biotechnological Applications of Cold-Adapted Organisms', Springer (1999), 1-16 [ISSN/ISBN: 3642636632] [DOI: 10.1007/978-3-642-58607-1] |
| Related plasmid reference: | De Wet et al., Mol. Cell. Biol. 7 (1987), 725-737 [PMID: 3821727] |
| 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 + tetracycline (10 μg/ml) |
| Cultivation temperature: | 37°C |
| Biosafety level: | L1 |
| Cultivation remark: | - |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pJBtrcLUC (LMBP 3134) is available at BCCM/GeneCorner. This plasmid was deposited by K. Keymeulen and Prof. Dr E. Remaut and was published in Remaut et al., 1999. |
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.