Patent ID: 8394610

Claim:
A method for constructing an operon comprising translationally coupled genes comprising: A) constructing a DNA fragment comprising an excisable gene coding for a selectable marker flanked by intercistronic regions, wherein a first intercistronic region is located upstream of the excisable gene, and said first intercistronic region comprises a first termination codon of a first gene to be translationally coupled, and a first SD-sequence, and a first initiation codon, wherein the first SD-sequence is located upstream of the first initiation codon and downstream of the first termination codon, wherein the first SD sequence can be partially overlapping with the first termination codon, and wherein a second intercistronic region is located downstream of the excisable gene, and said second intercistronic region comprises a second termination codon, a second SD-sequence, and a second initiation codon for a second gene to be translationally coupled, wherein the second SD-sequence is located upstream of the second initiation codon and downstream of the second termination codon, wherein the second SD sequence can be partially overlapping with the second initiation codon; B) integrating said DNA fragment between said genes to be translationally coupled using homologous recombination; and C) excising the gene coding for the selectable marker by a site-specific recombination system, and as a result of said excising, an ORF coding for a peptide results between the genes; wherein said intercistronic regions and said genes are from bacteria belonging to the genus Escherichia.