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GeneCorner plasmid details

Last data update: 24 January 2024 16:39 CET

Plasmid name: pSCGAL1MF2 (LMBP 2154)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p2154.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Saccharomyces cerevisiae α-mating factor 1 gene (MFα1); prepro secretion signal sequence (ppMF)
Promoter: Saccharomyces cerevisiae galactokinase promoter (GAL1)
Escherichia coli lac operon promoter
Ribosome
binding site:
-
Terminator: -
Selection marker: Ampicillin (amp)
Replicon: Escherichia coli plasmid pMB1 origin
Host range: Escherichia coli
Parental clone: pSCGAL1MF1; pUC19
Further information: The plasmid was constructed as follows: the HindIII-ScaI fragment of pSCGAL1MF1 (containing the prepro mating factor, the GAL1 and lac promoters and the Eco-pMB1 origin), was ligated to the ScaI-HindIII fragment of pUC19 (containing part of the amp gene and the lacZα gene). This construction created new expression sites behind the prepro sequence of the mating factor gene.
Expression sites: AccI, BspMI, HindIII, KpnI, SalI, SphI, Sse8387I and StuI, which are all unique.
Attention: the vector part of the plasmid is probably not entirely correct: the restriction sites were not all experimentally verified.
When cloning a fragment downstream from the lac promoter it may be advisable to use lacI(q) strains in order to prevent fortuitous expression of a possibly noxious polypeptide.
The nucleotide sequence at the 3' end of the GAL1 promoter is not exactly known.
EMBL Accession number: -
Latest sequence update: 22/03/1990
Authenticity test: The plasmid still needs to be subjected to the authenticity test.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Dr K. De Sutter(1) (2) and Prof. Dr R. Contreras(1) (2). It was constructed by W. Lammerant(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
Other culture collection numbers: -

Refer in your Materials and Methods:

pSCGAL1MF2 (LMBP 2154) is available at BCCM/GeneCorner. This plasmid was deposited by Dr K. De Sutter and 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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