GlgB and glgB-mutated strains in their E. coli DH5α genomic context, and the scar sequence left following in situ mutation. A. The five essential glycogen metabolism genes are organized into a single transcriptional unit as glgBXCAP in E. coli DH5α (Montero et al. ). For GlgB in E. coli DH5α, four domains have been identified, which are N-terminus, CBM48, α-amylase, and C-terminus. B. P1 and P2 (italic letters) were primers for amplifying linear PCR products from plasmid pKD4. Underlined sequence was FRT site with a palindromic sequence (blue sequences), separated by 12 asymmetric nucleotides. Red italic sequence was a ribosome-binding site and the green italic letters represented start codon for downstream gene expression.