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GeneCorner plasmid details

Last data update: 24 January 2024 16:39 CET

Plasmid name: pSCGAL1MF1 (LMBP 2116)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p2116.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:
Ribosome binding site (RBS) of the Escherichia coli lac Z gene (lacZ)
Terminator: -
Selection marker: Ampicillin (amp)
Replicon: Escherichia coli plasmid pMB1 origin
Host range: Escherichia coli
Parental clone: pSCTPIMF1; pSP64GAL1m
Further information: This plasmid was derived from pSCTPIMF1 as follows: the SalI-EcoRI fragment of pSP64GAL1m (containing the galactokinase (GAL1) promoter) was inserted into the XhoI and EcoRI sites of pSCTPIMF1.
Expression sites: HindIII and StuI.
The BamHI site at the 3' end of GAL1 is lost; the sequence in this region is not exactly known.
ATTENTION: the vector part of the plasmid is probably not correct: restriction sites were not experimentally verified.
Because of the presence of the lac operator, Lac+ E. coli host cells turn blue on X-Gal.
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.
Other name of the plasmid is pSCGal1MF2.
EMBL Accession number: -
Latest sequence update: 13/02/1990
Sequence detail:
Nucleotide sequence at the end of the prepro sequence:

   ---- ppMF ---->|
    80          83|       |
5' GTA.TCT.TTG.GAT.AAA.AGG|CCT.CTT.GAA.GCT.TGG 3'
   Val Ser Leu Asp Lys Arg|Pro Ala Glu Ala Trp
                         *|*** **
                       StuI         HindIII

*: mutated nucleotide
|: Cutting site of the alpha mating factor Kex2 endoprotease.
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 J. Demolder(1) and Prof. Dr R. Contreras(1).
(1) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: Demolder et al., Gene 111 (1992), 207-213 [PMID: 1541399]
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 DH5α
Host reference: Focus 8 (1986), 9
Related host reference: Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660]
Rodriguez-Quinones et al., Focus 15 (1993), 110-112
Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791] [DOI: 10.1016/s0022-2836(83)80284-8]
Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187]
Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051]
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:

pSCGAL1MF1 (LMBP 2116) is available at BCCM/GeneCorner. This plasmid was deposited by J. Demolder and Prof. Dr R. Contreras and was published in Demolder et al., 1992.

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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