Last data update: 18 January 2021 04:24 CET
Plasmid name: pSCGAL1-L (LMBP 4914)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner core plasmid|
(View with Genome Compiler)
|Promoter:||Saccharomyces cerevisiae galactokinase promoter (GAL1)|
|Terminator:||Saccharomyces cerevisiae 2 micron plasmid (2μ) FLP terminator
Saccharomyces cerevisiae iso-1-cytochrome C terminator (CYC1)
|Selection marker:||Ampicillin (amp)
|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.
|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
|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)|
|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.