Restriction digests of the clone give the following sizes (kb): EcoRI/HindIII--6.0, 1.6, 0.6; HindIII--8.4; BamHI--8.4; PstI--3.7, 3.4, 1.3; BglII--7.4, 1.1.
A marker addition/transplacement vector.
Non-integrants grow poorly selecting for Ura3+ (pinpoints at 5 days) while integrants grow well because the SUP11 copy number is maintained at one. If there are no integrants on a transformation plate, streak onto -URA plates and pick papillae.
Constructed by inserting the equivalent of 2 genomic EcoRI/HindIII fragments (1.5 kb, centromere proximal and 0.8 kb, 3' end) from the TRP1 region into YRp14.
Represents the deletion of a 0.6 kb EcoRI/HindIII fragment containing the promoter and the 5' portion of the gene.
Media
ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°C
References
Sikorski RS, Hieter P. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics 122: 19-27, 1989. PubMed: 2659436
