Tet-On3G: cutting CMV promoter out of pCMV-Tet3G - (Dec/12/2013 )
Hi,
I'm going to use the Tet3G coding sequence from pCMV-Tet3G and link it to another promoter.
Do anybody have experience in excising the CMV promoter sequence from the pCMV-Tet3G plasmid - and replacing it with another promoter. And then transform it in competent cells and subsequently purify the plasmid DNA.
This way I can make the restriction sites match and do not need to do multiple subclonings steps.
Thanks
-dsjensen-
The easiest way to generate compatible ends is to design primers with RE sites on the 5' end, then do a PCR of the site of interest, digest, and clone.
-bob1-