Hi,

If your gene is from eukaryote so you have to clone its cDNA. Then I must say , yes, you must include the start and stop codons for proper expression.

Be successful

Thank you so much. I have seen people exclude stop codon while designing primers for amplifying cDNA as they say there is an alpha galactosidase coding sequence downstream of the MCS in pGADT7. They also comment that further downstream of the MCS and alpha galactosidase, there is ultimately the ADH terminator.

I would be grateful if you could share your views in this regard also. I am attaching the map of pGADT7 for your reference.

Thanks and Regards