I'm trying to express a gene in P.p. with pGAPZalpha B. I though to linearize with BglII, which is at the begining of pGAP, but I have other BglII restriction sites in my gene of interest. So, I was wondering if cutting with BamHI, at the end of AOX I terminator could work, directing the integration in the AOX I loci. Invitrogen pGAP Manual recomend to cut with AvrII or BspHI, I could use any of them to but I'm not so sure if disrupting pGAP promoter I can have then functional expressing cassettes. Please, if anyone have tryed with success such alternatives before I will appreciate a lot your comments. Thank you very much in advance.