Hi everybody
I am trying to integrate a construct into the URA3 locus. I have two
constructs containing a partial deletion of that gene. The constructs
have either LEU or HIS selection. Upon cutting with a restriction enzyme
(the sites are different for the two plasmids) in the URA3 locus and
transformation into my URA3+ HIS- LEU- strain and selecting on LEU or
HIS plates, I get no colonies. As a control, I am using the same LEU
integration vector with a different locus cloned into it to integrate,
and this one gives me many (100s) colonies. Nobody I know has ever tried
integrating into the URA3 locus. If you do and you read this, please
email me with advice. I would greatly appreciate it. Thank you in
advance,
Amine
