M2660 [URA3 --> LYS2 converter] plasmid in E. coli (ATCC® 87559)

Applications: YI-type (integrating) shuttle vectorhost modificationmarker swap vector URA3 --> LYS2  /  Depositors: DJ Stillman

Permits and Restrictions

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Designations M2660 [URA3 --> LYS2 converter] plasmid in E. coli
Depositors DJ Stillman
Biosafety Level 1
Vector Information
Size (kb): 8.308
Vector: M2660 (plasmid)
Construction: pUC18
Marker(s):ampR LYS2
Construct size (kb): 8.308
Features: gene disruption cassette: ura3::LYS2
marker(s): ampR
replicon: pMB1
restriction site: HindIII
Applications
YI-type (integrating) shuttle vector
host modification
marker swap vector URA3 --> LYS2
Comments
Restriction digests of the clone give the following sizes (kb): BglII--4.3, 4.1; HindIII--5.6, 2.8; HpaI--8.5; PvuI--5.1, 2.4, 0.9.
To inactivate URA3, transform with HindIII-digested plasmid (5.57 kb) and select for Lys+.
A marker swap (URA3->LYS2) vector for altering a Saccharomyces cerevisiae host by a one-step gene disruption.
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.
Constructed by replacing an internal EcoRV-PstI fragment of URA3 with a 4.6 kb PvuII-PstI fragment containing LYS2. URA3 and LYS2 are in the same orientation.
Growth Conditions
Temperature: 37.0°C
References

Voth WP, et al. New 'marker swap' plasmids for converting selectable markers on budding yeast gene disruptions and plasmids. Yeast 20: 985-993, 2003.

Related Products
component of:ATCC 87561
Shipped frozen
Product Sheet
Product Sheet
Product Sheet
Attachments Vector Map for 87559