IRES in expression vector - (Oct/03/2014 )

HI 

I am going to construct mammaian expression vectors for two independent genes and I need to use an IRES between them. I am not sure if my second gene  should be just after the IRES or it is possible to have a few nucleotides between IRES and second gene. and also do I need to have an ATG start codon for my second gene?

thanks

This may be too late, but yes, you can have a number of bases between an IRES and the ATG, and you do need an ATG.