Last data update: 24 January 2024 16:39 CET
Plasmid name: pIRES-LUC (LMBP 3472)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p3472.gb
(View with Genome Compiler) p3472.txt p3472.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Photinus pyralis (firefly) luciferase gene (LUC); mutated coding region (LUCm; luc(+)) |
| Promoter: | Phage T7 gene 10 promoter (T7g10) Phage T3 promoter Escherichia coli lac operon promoter |
| Ribosome binding site: |
Internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV) polyprotein gene |
| Terminator: | Simian virus 40 polyadenylation signal (SV40 polyA) |
| Selection marker: | Ampicillin (amp) |
| Replicon: | Escherichia coli plasmid pMB1 origin Phage f1 origin |
| Host range: | Escherichia coli Mammalian cells |
| Parental clone: | pIRES-lacZ; pGL3-Basic |
| Further information: | This phasmid was derived from pIRES-lacZ by removing the lacZ gene as an NcoI-XhoI fragment and replacing it by an NcoI-SalI fragment from pGL3-Basic, containing the mutated P. pyralis luciferase gene. The IRES-LUCm insert allows to obtain bicistronic or polycistronic transcripts in mammalian cells, using the mutated firefly luciferase gene as an efficient reporter gene. LUCm contains several silent mutations to improve codon usage, as well as mutations resulting in a change of amino acids to eliminate potential glycosylation and ATF sites (amino acid positions 50 and 119) and to remove the peroxisome targeting sequence (amino acid positions 548-550). A complete description of all modifications of the luciferase gene can be found in Sherf et al. (1994). The internal ribosome entry site allows ribosomes to enter and bind functionally to an mRNA independent of a cap site. When cloning a fragment downstream from the lac promoter it may be advisable to use lacI(q) strains in order to prevent fortuitous expression of a possibly noxious polypeptide. The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727341.1. The nucleotide sequence of the 5' untranslated region of the EMCV polyprotein gene, containing the IRES sequence, corresponds with the EMBL Nucleotide Sequence Database accession number M81861.1. Other name of the plasmid is pBlue-IRESluc. |
| EMBL Accession number: | M81861.1, view at EMBL, GenBank, DDBJ LT727341.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 01/04/1996 |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: AvaI/ClaI, BamHI, BglI, EcoRI, HindIII and NcoI. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Dr J. Tavernier(1). (1) Roche Research Ghent, Gent, Belgium |
| Plasmid reference: | - |
| Related plasmid reference: | Sherf et al., Promega Notes Magazine 49 (1994), 14-21 |
| Restricted distribution: | - BCCM MTA |
| Distributed as: | H/P active culture or plasmid DNA |
| Host for distribution: | Escherichia coli K12 DH5α |
| Host reference: | Focus 8 (1986), 9 |
| Related host reference: | Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660] Rodriguez-Quinones et al., Focus 15 (1993), 110-112 Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791] [DOI: 10.1016/s0022-2836(83)80284-8] Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187] Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051] |
| Cultivation medium: | LB-Lennox + ampicillin (100 μg/ml) |
| Cultivation temperature: | 37°C |
| Biosafety level: | L1 |
| Cultivation remark: | - |
| Other culture collection numbers: | - |
| Related website: | http://www.iresite.org/IRESite_web.php?page=view&entry_id=140 |
Refer in your Materials and Methods: |
pIRES-LUC (LMBP 3472) is available at BCCM/GeneCorner. This plasmid was deposited by Dr J. Tavernier. |
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.