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Kozak sequences..


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#1 khyros

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Posted 21 October 2009 - 05:09 AM

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.

#2 bob1

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Posted 21 October 2009 - 03:41 PM

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




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