so do u intend to make a fusion protein?
if so then use the last 20 amino acids , removing the stop codon and
aproximately 80 to 100 basepairs down the 3'UTR and use a downstream primer
of 60 basepairs.
this way i have constructed atleast 20 different tags and they all worked.
so good luck
Franco.
> -----Original Message-----
> From: Marjon A.van Slegtenhorst [mailto:MA_vanSlegtenhorst at fccc.edu]
> Sent: Wednesday, July 03, 2002 2:05 PM
> To: yeast at net.bio.net> Subject: integration of GFP tag
I want to design primers to intergrate a GFP tag C-term of my gene from
the pFA6a-GFP(S65T)-kanMX6 plasmid, but I'm not sure how far the two
gene specific sequences should be apart to get a successful integration.
I want to delete as little as possible from the endogenous 3' region...
Thanks for the advise..
Marjon
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