Restructuring\na Complex Genetic Function on Episomal\nVectors in Escherichia coli

Genetic functions have evolved over long timescales and\ncan be\nencoded by multiple genes dispersed in different locations in genomes,\nand although contemporary molecular biology enables control over single\ngenes, more complex genetic functions remain challenging. Here, we\nstudy the restructuring and mobilization of a complex genetic function\nencoded by 10 genes, originally expressed from four operons and two\nloci on the Escherichia coli genome.\nWe observe subtle phenotypic differences and reduced fitness when\nexpressed from episomal DNA and demonstrate that mutations in the\ntranscriptional machinery are necessary for successful implementation\nin different bacteria. The work provides new approaches for advanced\ngenome editing and constitutes a first step toward modularization\nand genome-level engineering of complex genetic functions.