Hi, @ALL

In my fusion cloning construct, I noticed that instead of the expected stop codon 'TAA' at the end of the EGFP tag, the sequence includes an extra nucleotide"C", resulting in 'TCAA'. What could be the possible reasons for this unexpected insertion of an extra (C) in the stop codon? Could it be due to primer design, cloning artifacts, frame shifts, or vector-specific sequences? I would appreciate any insights on how such anomalies might occur during the cloning process. Thanks