This material is cited in a US or other Patent and may not be used to
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Restriction digests of the clone give the following sizes (kb): BamHI--15.0; BglII--15.0; XhoI--9.4, 6.0; EcoRI--13.0, 2.1; KpnI--13.0, 2.2.
Saccharomyces cerevisiae containing a single copy of the neomycin phosphotransferase gene are sensitive to G418; resistance to 400 ug/ml is expressed if there are several copies per cell.
The promoter and terminator sequences were derived from the phosphoglycerate kinase gene.
The G418-resistance segment includes a 0.7 kb region of the PGK promoter and a 1.5 kb BglII/BamHI fragment from Tn5. The BamHI site is between the G418R and the PGK terminator.
Media
ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°C
References
Patel AC, Ratzkin BJ. Materials and methods for controlling plasmid copy number and stability. US Patent 4,935,350 dated Jun 19 1990