pUT11 [URA3 --> TRP1 converter] (ATCC® 87553)

Applications: YI-type (integrating) shuttle vectorhost modificationmarker swap vector URA3 --> TRP1  /  Depositors: FR Cross

Permits and Restrictions

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Designations pUT11 [URA3 --> TRP1 converter]
Depositors FR Cross
Biosafety Level 1
Host
Distribution host: Escherichia coli HB101 (ATCC 33694)
Vector Information
Size (kb): 6.6999998092651370
DESCRIPTION OF VECTOR:
Intact vector size: 6.700
Type of vector: plasmid
Cloning sites:
Polylinker sites:
Construction: pUC19
Host range: Saccharomyces cerevisiae; Escherichia coli
Features (with orientation and position when available):
replicon: pMB1
marker(s): ampR, <-
restriction site: EcoRI, SmaI
gene disruption cassette: ura3::TRP1/kanR
restriction site: SmaI
Vector: pUT11 (plasmid)
Construction: pUC19
Marker(s):TRP1,ampR,kanR
Construct size (kb): 6.699999809265137
Features: gene disruption cassette: ura3::TRP1/kanR
marker(s): ampR
replicon: pMB1
restriction site: EcoRI, SmaI
restriction site: SmaI
Applications
YI-type (integrating) shuttle vector
host modification
marker swap vector URA3 --> TRP1
Comments
Restriction digests of the clone give the following sizes (kb): HindIII--4.6, 2.0; SmaI--3.8, 2.7.
To convert the host phenotype from URA3 to TRP1, transform with the SmaI digested vector and select for Trp+ 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 TRP1 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 TRP1 and kanR coding sequences. TRP1 and URA3 are in the same 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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component of:ATCC 87561
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Shipping Information Distributed: freeze-dried
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