expression of antisense seq in pYES1.2

Ken Howe howe at DARWIN.UCSC.EDU
Sun Feb 25 13:53:49 EST 1996


Hey fellow Netters,

I've been working a bit with pYES1.2 (GAL promoter, CYC1 termination in a 
Kananmycin, URA3, ColE1/2micron plasmid) and discovered that my insert 
was being expressed in both orientations.  This is supposed to be a 
promoterless insert, so before I do any further analysis to see if 
there's a cryptic promoter present, I'd like to see if anyone has had any 
similar experiences with this plasmid and whether they saw antisense 
expression in the sense orientation or sense expression in the opposite 
orientation.  In other words, has anyone observed either transcription 
initiation within the CYC1 termination signal (and extending towards the 
GAL promoter) or of run-through transcription from the URA3 gene located 
about 2kb upstream?

Any comments would be helpful.

Thanks.

Ken Howe
Center for the Molecular Biology of RNA
University of California
Santa Cruz, CA  95064





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