Last data update: 24 January 2024 16:39 CET
Plasmid name: pEFlacZneoflox (LMBP 4478)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p4478.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Transposon Tn5 neomycin phosphotransferase gene (neo) Escherichia coli lac Z gene (lacZ); inactive fragment |
| Promoter: | Human elongation factor 1α promoter (EF1α) Simian virus 40 early promoter (SV40 early) |
| Ribosome binding site: |
- |
| Terminator: | Human granulocyte macrophage colony stimulating factor polyadenylation signal (GM-CSF polyA) |
| Selection marker: | Ampicillin (amp) Neomycin (neo; G418) |
| Replicon: | Escherichia coli plasmid pMB1 origin Simian virus 40 bidirectional origin (SV40) Phage f1 origin |
| Host range: | Escherichia coli Mammalian cells; SV40 permissive cells |
| Parental clone: | pRFlacZ |
| Further information: | The plasmid can be used to test cre-expressing vectors in transient cotransfection assays or to generate reporter gene cell lines. In cotransfection experiments the cre expression vector and the reporter gene plasmid should be used in equimolar amounts. The lacZ gene in pEFlacZneoflox is inactive due to the insertion of a loxP flanked neomycin resistance gene. The lacZ functionality will be restored through the Cre-mediated excision of the neomycin resistance gene. Therefore neomycin (G418) has to be omitted in the cell culture medium during and after cre expression. The expressed β-galactosidase can subsequently be detected by X-Gal staining. |
| EMBL Accession number: | - |
| Latest sequence update: | 15/05/2002 |
| 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 Prof. Dr W. Muller(1). (1) Faculty of Life Sciences, University of Manchester, Manchester, United Kingdom |
| Plasmid reference: | - |
| 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 |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pEFlacZneoflox (LMBP 4478) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr W. Muller. |
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.