Last data update: 24 January 2024 16:39 CET
Plasmid name: pSCTyGAL1-L (LMBP 4913)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p4913.gb
(View with Genome Compiler) p4913.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
- |
| Promoter: | Saccharomyces cerevisiae galactokinase promoter (GAL1) |
| Ribosome binding site: |
- |
| Terminator: | Saccharomyces cerevisiae 2 micron plasmid (2μ) FLP terminator Saccharomyces cerevisiae iso-1-cytochrome C terminator (CYC1) |
| Selection marker: | Ampicillin (amp) LEU2; auxotrophic |
| Replicon: | Escherichia coli plasmid pMB1 origin |
| Host range: | Escherichia coli Saccharomyces cerevisiae; leu2(-), integrative |
| Parental clone: | pUT332; pSP64Gal1m; pEMBLyex4; pUD208; pFV17 |
| Further information: | The plasmid was constructed as follows: 1) The Saccharomyces cerevisiae 2 micron plasmid origin (2μ) and the S. cerevisiae uracil 3 (URA3) marker gene were removed from pUT332 by successive digestions with ClaI and BglII. 2) The BamHI-HindIII fragment from pSP64Gal1m, containing the S. cerevisiae galactokinase promoter (GAL1), was ligated into the BglII-HindIII opened plasmid. 3) The XbaI-FspI fragment from pEMBLyex4, containing the S. cerevisiae 2μ FLP terminator, was inserted into this XbaI-HindIII opened plasmid (HindIII was blunted with Klenow DNA polymerase). 4) The resulting YIpUT construct was KpnI-AatII opened (blunted with T4 DNA polymerase) and the EcoRI-BsaAI Ty1 δ fragment (EcoRI was blunted with Klenow DNA polymerase) from pUD208 was inserted. 5) Finally the BsaAI-BsrGI (BsrGI was blunted with Klenow DNA polymerase) from pFV17, containing the LEU2 coding sequence, was inserted into the BamHI (blunted with Klenow DNA polymerase) opened plasmid, resulting in pSCTyGAL1-L. pSCTyGAL1-L is designed for integrative expression in S. cerevisiae. After XhoI linearisation in its Ty1 δ element, multicopy genomical integration leads to stable expression. Other name of the plasmid is YIpUTyL. |
| EMBL Accession number: | - |
| Latest sequence update: | 30/06/2004 |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: AgeI/XmnI, BstEII/XhoI, EcoRI, HindIII, HindIII/XbaI, KpnI/XbaI, MluI, PstI and XhoI. The XmnI site at position 3569 could not be experimentally confirmed. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Prof. Dr R. Contreras(1) (2). (1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium (2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | Reekmans et al., FEMS Yeast Res. 5 (2005), 711-725 [PMID: 15851100] [DOI: 10.1016/j.femsyr.2004.12.010] |
| 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 + ampicillin (100 μg/ml) |
| Cultivation temperature: | 37°C |
| Biosafety level: | L1 |
| Cultivation remark: | - |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pSCTyGAL1-L (LMBP 4913) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr R. Contreras and was published in Reekmans et al., 2005. |
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.