pHU10 [HIS3 --> URA3 converter] (ATCC® 87544)

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

Permits and Restrictions

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Designations pHU10 [HIS3 --> URA3 converter]
Depositors FR Cross
Biosafety Level 1
Host Escherichia coli HB101
Vector Information
Construct size (kb): 6.60
Type of vector: Phagemid
Host range: Saccharomyces cerevisiae, Escherichia coli

Features (with orientation and position when available):
replicon: f1, →
marker(s): cmlR, ←
replicon: pMB1
restriction site: XhoI, SalI
Applications
YI-type (integrating) shuttle vector
host modification
marker swap vector HIS3 --> URA3
Comments
Restriction digests of the clone give the following sizes (kb): HindIII--6.5; HindIII/PstI--5.0, 1.1, 0.5.
To convert the host phenotype from HIS3 to URA3, transform with the SmaI+XhoI digested vector and select for Ura+ 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 the URA3 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 HindIII fragment of HIS3 with a SmaI fragment containing the URA3 and kanR coding sequences. URA3 and HIS3 are in the opposite orientation.
Media ATCC Medium 1065 (see below) plus kanamycin (50.0 mcg/ml) plus chloramphenicol (25.0 mcg/ml) ATCC Medium 1065: Tryptone (Difco 0123), 10.0 g Yeast Extract (Difco 0127), 5.0 g NaCl, 10.0 g Distilled water, 1.0 L
Growth Conditions
Temperature: 37°C
References

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

Related Products
Component of: ATCC 87561
Shipped freeze-dried
Product Sheet
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Product Sheet
Attachments Vector map for 87544