pUL9 [URA3 --> LEU2 converter] (ATCC® 87552)

Applications: YI-type (integrating) shuttle vectorhost modificationmarker swap vector URA3 --> LEU2  /  Depositors: FR Cross

Permits and Restrictions

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Designations pUL9 [URA3 --> LEU2 converter]
Depositors FR Cross
Biosafety Level 1
Vector Information
Size (kb): 7.5999999046325680
Vector: pUL9 (plasmid)
Construction: pUC19
Marker(s):LEU2,ampR,kanR
Construct size (kb): 7.599999904632568
Features: gene disruption cassette: ura3::kanR/LEU2
marker(s): ampR
replicon: pMB1
restriction site: SmaI, XbaI
restriction site: SmaI, XbaI, HindIII
Applications
YI-type (integrating) shuttle vector
host modification
marker swap vector URA3 --> LEU2
Comments
Restriction digests of the clone give the following sizes (kb): SmaI--4.3, 3.2; EcoRI--4.5, 2.9.
To convert the host phenotype from URA3 to LEU2, transform with the SmaI digested vector and select for Leu+ 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 LEU2 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 LEU2 and kanR coding sequences. LEU2 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
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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