Last data update: 09 August 2020 04:16 CEST
Plasmid name: pGAL1PSiST-1 (LMBP 4177)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner core plasmid|
(View with Genome Compiler)
|Cloned DNA:||Human interferon β gene (IFNB, IFNB1); stuffer fragment|
|Promoter:||Candida albicans galactokinase 1 promoter (CaGAL1)
Escherichia coli lac operon promoter
|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.
|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)|
|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.