How to exchange CMV for another promoter - (Nov/28/2005 )

I want to make a plasmid with some tag and an insertion of my gene but using my promoter exchanged with the original CMV, such as: promoter A+ATG+FLAG+gene A+stop codon. When i look for expression vectors with some tag and MCS sites, they always contain a CMV promoter, but i cannot find commonly used enzymes to cut off and exchange it for my promoter, does anyone have any idea about this? And is it possible to make such a plasmid construct? How to make it efficiently? Thanks.

--Jason

I think I suggested this before, but can't you clone your putative promoter+gene into the vector in the orientation opposite to the CMV promoter?

Or use a vector with a promoterless copy of an easily assayable reporter gene (luciferase? lacZ? GFP? antibiotic resistance?). We use a promoterless xylE gene -- if the fragment cloned upstream of it has promoter activity, colonies sprayed with cathecol turn yellow -- see photographs in Nature 414, 555-558 (29 November 2001).