Restriction digests of the clone give the following sizes (kb): HindIII--4.6, 2.0; SmaI--3.8, 2.7.
To convert the host phenotype from URA3 to TRP1, transform with the SmaI digested vector and select for Trp+ transformants.
Some combinations of marker swap plasmids and target locus may result in relatively high reversion rates. In most but not all cases the frequencies of successful convertants are greater than 30%.
A marker swap vector designed to change the S. cerevisiae host phenotype by one-step gene disruption of the URA3 gene with the TRP1 and kanR markers.
When swapping markers on an episomal plasmid, appropriate phenotype may result from loss of the plasmid unless a second selectable or scorable marker is used to ensure plasmid maintenance.
Vector was constructed by replacing an internal StuI fragment of URA3 with a SmaI fragment containing the TRP1 and kanR coding sequences. TRP1 and URA3 are in the same orientation.
Media
ATCC® Medium 1948: LB medium (ATCC medium 1065) with 50 mcg/ml ampicillin and 20 mcg/ml kanamycin
