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Tet-On3G: cutting CMV promoter out of pCMV-Tet3G - (Dec/12/2013 )



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.




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.