Restriction digests of the clone give the following sizes (kb): EcoRI/ClaI--6.1, 1.6; SalI--5.9, 1.8; EcoRI/HindIII--6.2, 1.5; HindIII--7.7.
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.
A 0.6 kb EcoRI/ClaI fragment (blunt-ended) was deleted from a 2.2 kb genomic SalI/XhoI fragment having a SalI linker. The SalI fragment was cloned into YRp15.
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
