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serema2003 06-27-2012 12:01 PM

Protein expression fused with GFP
 
Hi
sorry if my question seems very basic! I really do nott have access to my protocols now and I do not clearly remember what we did before.Protein has not been my area of expertise.
I want to clone a known cDNA with Age1,Nhe1 upstream of IRES sequence so my final protein be expressed in fusion with GFP located at 3prime.here is the address of vector 26670 at addgene

That is Promoter(Synapsin)+cDNA+IRES+GFP+wpre+ltr+PA terminator

Since my full length cDNA is already cloned in another vector I just need two primer with Age1 and Nhe1 sites to amplify the fragment and clone into that aforementioned vector.the problem is that I'm not sure where these primers should start and end?I mean the first of primer should be ATG of cDNA and the end before pa signal or after that in cDNA???
plz help me guys


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