If I use Gateway® cloning system for 5′ fusion protein, the recombination site (attB) and the Multiple Cloning Site (MCS) in the ORFEXPRESS™ Shuttle Clone will become coding sequence. How many amino acids will be added?

Answer: There are fourteen amino acids between the fusion peptide (or N-terminal Tag) and the ORF encoded protein. These fourteen amino acids are, from N-terminus to C-terminus, Thr Ser Leu Tyr Lys Lys Ala Gly Leu Gly Gly Val Arg Thr. Eight amino acids (Thr Ser Leu Tyr Lys Lys Ala Gly) are encoded by the attB recombination site and six (Leu Gly Gly Val Arg Thr) are encoded by the sequences between attB and the start codon, ATG.