pHL3 [HIS3 --> LEU2 converter] (ATCC® 87542)

Applications: YI-type (integrating) shuttle vectorhost modificationmarker swap vector HIS3 --> LEU2  /  Depositors: FR Cross

Permits and Restrictions

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Designations pHL3 [HIS3 --> LEU2 converter]
Depositors FR Cross
Biosafety Level 1
Vector Information
Size (kb): 7.6999998092651370
Vector: pHL3 (phagemid)
Construction: pBC KS+
Marker(s):LEU2,cmlR
Construct size (kb): 7.699999809265137
Features: gene disruption cassette: his3::LEU2
marker(s): cmlR
replicon: f1
replicon: pMB1
restriction site: ApaI, SalI
restriction site: PstI
Applications
YI-type (integrating) shuttle vector
host modification
marker swap vector HIS3 --> LEU2
Comments
To convert the host phenotype from HIS3 to LEU2, transform with the ApaI+PstI 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 HIS3 gene with a LEU2 marker.
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 BglII fragment of HIS3 with a 2.7 kb BglII fragment containing the LEU2 coding sequence. LEU2 and HIS3 are in the same orientation.
Media Medium 1675: LB medium (ATCC medium 1065) with 10 mcg/ml Chloramphenicol
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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