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

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

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