The accD gene encoding the β-subunit of acetyl-CoA carboxyltransferase, the accA gene encoding the α-subunit of acetyl-CoA carboxyltransferase, and the accBC operon encoding biotin carboxyl carrier protein and biotin carboxylase were amplified by PCR from MG1655 genomic DNA with their putative upstream RBS using primers 7 and 8, 9 and 10, and 11 and 12, respectively.