Kozak sequences.. - (Oct/21/2009 )

Hello, I'm starting a new part of my project that involves the overexpression of an enzyme that I have previously studied, silenced, and made chemical inhibitors for.

I haven't done much cloning in the past (only "posh" cloning using the invitrogen Gateway system so I lack experience in old school molecular genetics that I now need to do since I'm doing my phd for a state-funded institute).

We bought the cDNA from the MGC clone library, and it comes in a pCMV-sport6 vector. Since I need to make stable cell lines, I will transfer the insert with the cDNA into a pCDNA3.1 vector.

I see that I need a kozak sequence upstream the start codon. I checked and I have the following:

gtcGCCatgG

is this GCCATGG sequence ok for a Kozak? I would assume so but just wanted to make sure

Thanks a lot.

Usual kozak is GCCACCATG, shorter and longer versions work, but with lower efficiency.