Last data update: 24 January 2024 16:39 CET
Plasmid name: pSCGAL1-L (LMBP 4914)
New search | Print data sheet |
Price category: | Cat. 1 (cf. price list) |
Status: | GeneCorner core plasmid |
GeneCorner sequence: |
p4914.gb
(View with Genome Compiler) p4914.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; 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), resulting in YIpUT. 4) Finally the BsaAI-BsrGI fragment (BsrGI was blunted with Klenow DNA polymerase) from pFV17, containing the S. cerevisiae leucine 2 (LEU2) coding sequence, was inserted into this SmaI-Eco47III opened YIpUT plasmid, resulting in pSCGAL1-L. pSCGAL1-L is designed for integrative expression in S. cerevisiae. After BstEII linearisation in its LEU2 element, single-copy genomical integration leads to stable expression. Other name of the plasmid is YIpUTL. |
EMBL Accession number: | - |
Latest sequence update: | 30/06/2004 |
Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: AgeI/XmnI, BamHI/HindIII, BstEII/XhoI, HindIII/XbaI, MluI, PstI, SmaI/XbaI and XhoI. The XmnI site at position 3664 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: | - |
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: |
pSCGAL1-L (LMBP 4914) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr R. Contreras . |
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.