pUH7 [URA3-->HIS3 converter] [URA3 --> HIS3 converter] (ATCC® 87551)

Applications: YI-type (integrating) shuttle vectorhost modificationmarker swap vector URA3 --> HIS3  /  Depositors: FR Cross

Permits and Restrictions

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Designations pUH7 [URA3-->HIS3 converter] [URA3 --> HIS3 converter]
Depositors FR Cross
Biosafety Level 1
Vector Information
Construct size (kb): 6.5
Applications
YI-type (integrating) shuttle vector
host modification
marker swap vector URA3 --> HIS3
Comments
Restriction digests of the clone give the following sizes (kb): EcoRI--6.4; SmaI--3.5, 2.8; XbaI--3.6, 2.7.
To convert the host phenotype from URA3 to HIS3, transform with the SmaI digested vector and select for His+ 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 HIS3 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 HIS3 and kanR coding sequences. HIS3 and URA3 are probably in the opposite orientation.
Media Medium 1948: LB medium (ATCC medium 1065) with 50 mcg/ml ampicillin and 20 mcg/ml kanamycin
Growth Conditions
Temperature: 37.0°C
References

Cross FR. 'Marker swap' plasmids: convenient tools for budding yeast molecular genetics. Yeast 13: 647-653, 1997. PubMed: 9200814

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