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GeneCorner plasmid details

Last data update: 15 November 2019 04:07 CET  


Name: pGAL1PSiST-1   Add to cart
Accession number: LMBP 4177
Sequence file: LMBP sequence: p4177.gb, p4177.txt     View with Genome Compiler
Circular map: p4177.pdf
Status: GeneCorner core plasmid
Cloned DNA: Human interferon β gene (IFNB, IFNB1); stuffer fragment
Promoter: Candida albicans galactokinase 1 promoter (CaGAL1)
Escherichia coli lac operon promoter
Ribosome
binding site:
-
Terminator: Saccharomyces cerevisiae iso-1-cytochrome C terminator (CYC1)
Selection marker: Ampicillin (amp)
Candida albicans URA3; auxotrophic
Replicon: Escherichia coli plasmid pMB1 origin
Host range: Escherichia coli
Candida albicans; ura3(-), integrative
Parental clone: pMAL2PHiET-3; YCp50SS; pGAL1PNiST-1
Further information: The plasmid was constructed as follows: the EcoRI-HindIII fragment from pMAL2PHiET-3, containing the human IFNβ coding sequence flanked by SfiI and NotI restriction sites, was exchanged by the EcoRI-HindIII fragment from YCp50SS, containing the human IFNβ coding sequence flanked by two SfiI restriction sites. In this intermediate pMAL2PSiST-1 vector, the NaeI-SalI MAL2 promoter fragment was replaced by the FspI-XhoI GAL1 promoter fragment from pGAL1PNiST-1, resulting in pGAL1PSiST-1.
This integrative vector is useful for the construction of Candida albicans libraries.
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 of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727322.1.
The nucleotide sequence of the hIFNβ cDNA corresponds with the EMBL Nucleotide Sequence Database accession number V00546.1.
The nucleotide sequence of the CaURA3 DNA corresponds with the EMBL Nucleotide Sequence Database accession number X14198.1, except for three nucleotides (silent mutations) at position 135 (A->G), 141 (T->C) and 144 (A->G) of the coding sequence respectively. LMBP WILL ANALYSE THE CaURA3 SEQUENCE TO VERIFY POSSIBLE CORRESPONDENCE WITH AF109400.1.
EMBL Accession number: V00546.1, view at EMBL, GenBank, DDBJ
X14198.1, view at EMBL, GenBank, DDBJ
LT727322.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 28/08/2000
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BamHI, ClaI, EcoRV, HindII, HindIII and SfiI.

Although a dam(-) strain was transformed with the plasmid, the ClaI digestion was ambiguous.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Janssen Pharmaceutica N.V.(1). It was constructed by Dr J. Viaene(2).
(1) Beerse, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: De Backer et al., Nat. Biotechnol. 19 (2001), 235-241 [PMID: 11231556]
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: -

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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