Last data update: 24 January 2024 16:39 CET
Plasmid name: pCfB2225 (LMBP 9501)
New search | Print data sheet |
Price category: | Cat. 1 (cf. price list) |
Status: | GeneCorner core plasmid |
GeneCorner sequence: |
p9501.gb
(View with Genome Compiler) p9501.txt p9501.pdf |
Sequence analysis results Genecorner: |
- |
Cloned DNA: |
- |
Promoter: | Phage T7 gene 10 promoter (T7g10) Eremothecium gossypii translation elongation factor 1α promoter (TEF1α) |
Ribosome binding site: |
- |
Terminator: | Saccharomyces cerevisiae alcohol dehydrogenase terminator (ADH1) Saccharomyces cerevisiae iso-1-cytochrome C terminator (CYC1) Eremothecium gossypii translation elongation factor 1α terminator (TEF1α) Escherichia coli rrnB operon T1 terminator |
Selection marker: | Ampicillin (amp) Neomycin (neo; G418) |
Replicon: | Escherichia coli plasmid pMB1 origin |
Host range: | Escherichia coli Saccharomyces cerevisiae |
Parental clone: | pXII-2 |
Further information: | The plasmid contains two S. cerevisiae 2μ FLP Recombinase Target sites (FRT sites), two S. cerevisiae chromosome XII integration targeting sequences and two bacteriophage P1 Cre recombinase target sites (loxP sites), next to the USER cloning cassette (AsiSI and Nb.BsmI). The plasmid is part of the EasyClone integrative vector set and contains a loxP-flanked antibiotic resistance selection marker. This set allows simultaneous expression of multiple genes in S. cerevisiae with an option of recycling selection markers. The kanMX cassette contains the Tn903 neomycin coding sequence fused to transcriptional and translational control sequences from the filamentous fungus E. gossypii TEF1α gene, rendering yeast strains resistance to geneticin (G418). The plasmid confers no resistance to kanamycin in bacteria. This plasmid allows for integration of a selection marker on S. cerevisiae chromosome XII, in between two essential genetic elements. The integration site provides a good level of gene expression, has a minimum risk of spontaneous loop out, and does not impair growth. Other name of the plasmid is pXII-2-loxP-KanMXsyn. |
EMBL Accession number: | - |
Latest sequence update: | 04/12/2014 |
Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BglII/XhoI, HindIII, NcoI and SspI. |
Class: | Recombinant plasmid |
Type: | Plasmid |
History of deposit: | This plasmid was deposited by Prof. Dr I. Borodina(1). It was constructed by Dr. V. Stovicek(1). (1) The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Horsholm, Denmark |
Plasmid reference: | - |
Related plasmid reference: | Jensen et al., FEMS Yeast Res. 14 (2014), 238-248 [PMID: 24151867] [DOI: 10.1111/1567-1364.12118] Mikkelsen et al., Metab. Eng. 14 (2012), 104-111 [PMID: 22326477] [DOI: 10.1016/j.ymben.2012.01.006] |
Restricted distribution: | - Depositor's Material Transfer Agreement (DTU-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: | - |
Refer in your Materials and Methods: |
pCfB2225 (LMBP 9501) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr I. Borodina. |
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.