Restriction digests of the clone give the following sizes (kb): BglII--4.3, 4.1; HindIII--5.6, 2.8; HpaI--8.5; PvuI--5.1, 2.4, 0.9.
To inactivate URA3, transform with HindIII-digested plasmid (5.57 kb) and select for Lys+.
A marker swap (URA3->LYS2) vector for altering a Saccharomyces cerevisiae host by a one-step gene disruption.
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.
Constructed by replacing an internal EcoRV-PstI fragment of URA3 with a 4.6 kb PvuII-PstI fragment containing LYS2. URA3 and LYS2 are in the same orientation.
Growth Conditions
Temperature: 37.0°C
References
Voth WP, et al. New 'marker swap' plasmids for converting selectable markers on budding yeast gene disruptions and plasmids. Yeast 20: 985-993, 2003.
