Last data update: 24 January 2024 16:39 CET
Plasmid name: pCfB388 (LMBP 9502)
New search | Print data sheet |
Price category: | Cat. 1 (cf. price list) |
Status: | GeneCorner non-core plasmid |
Depositor's sequence: | p9502.gb |
Sequence analysis results Genecorner: |
- |
Cloned DNA: |
- |
Promoter: | Phage T7 gene 10 promoter (T7g10) Kluyveromyces lactis β-isopropylmalate dehydrogenase promoter (LEU2) |
Ribosome binding site: |
- |
Terminator: | Saccharomyces cerevisiae alcohol dehydrogenase terminator (ADH1) Saccharomyces cerevisiae iso-1-cytochrome C terminator (CYC1) Kluyveromyces lactis β-isopropylmalate dehydrogenase terminator (LEU2) |
Selection marker: | Ampicillin (amp) Kluyveromyces lactis LEU2; auxotrophic |
Replicon: | Escherichia coli plasmid pMB1 origin |
Host range: | Escherichia coli Saccharomyces cerevisiae; leu2(-), integrative |
Parental clone: | pCfB383 |
Further information: | The plasmid was constructed based on pCfB383 by replacing the direct repeats flanked K. lactis URA3 selection marker with a LoxP-flanked K. lactis LEU2 marker. The selection marker exchange was accomplished by uracil-specific excision reaction (USER). The plasmid contains two S. cerevisiae 2μ FLP Recombinase Target sites (FRT sites), two S. cerevisiae chromosome XI 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 auxotrophic selection marker. This set allows simultaneous expression of multiple genes in S. cerevisiae via genomic integration, with an option of recycling selection markers. This plasmid allows for integration of a selection marker on S. cerevisiae chromosome XI, 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 pXI-1-LoxP-KlLEU2. |
EMBL Accession number: | - |
Latest sequence update: | 04/12/2014 |
Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: HindIII/NheI, PstI, ScaI 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. N.B. Jensen(1). (1) The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Horsholm, Denmark |
Plasmid reference: | Jensen et al., FEMS Yeast Res. 14 (2014), 238-248 [PMID: 24151867] [DOI: 10.1111/1567-1364.12118] |
Related plasmid reference: | 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: |
pCfB388 (LMBP 9502) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr I. Borodina and was published in Jensen et al., 2014. |
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.