I want to clone a PCR product with added restriction sites into a pLenti vector under an EF1a promoter. The forward primer contains the start codon (ATG) of the gene of interest, is this enough to get the insert in frame? If not, how do I make sure that my insert is in frame? Is this even important, since a frameshift would occur between the start and the stop codons, right?
Thanks in advance for your reply,