Restriction digests of the clone give the following sizes (kb): BamHI--11.0, 0.5; PstI--6.1, 4.0, 1.2; BglII--10.0, 1.15; KpnI--11.5.
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 1.0 kb region that encompasses the entire coding region of HIS3 was deleted from a genomic EcoRI/SalI fragment (final size=5.1 kb) cloned into the EcoRI/SalI sites of YRp14.
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
Fasullo MT, Davis RW. Direction of chhromosome rearrangements in Saccharomyces cerevisiae by use of his3 recombinational studies. Mol. Cell. Biol. 8: 4370-4380, 1988. PubMed: 3054515
