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GeneCorner plasmid details

Last data update: 18 January 2021 04:24 CET

Plasmid name: pSCGAL1-L (LMBP 4914)

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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.

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