Postma ED, Dashko S, van Breemen L, Taylor Parkins SK, van den Broek M, Daran JM, Daran-Lapujade P.

Nucleic Acids Res. 2021 Feb 22;49(3):1769-1783. doi:10.1093/nar/gkaa1167

A supernumerary designer chromosome for modular in vivo pathway assembly in Saccharomyces cerevisiae.

Circular neochromosome assembled in vivo by homologous recombination of fragments containing short homology regions (60 bp). Includes “a centromere and autonomously replicating sequences (ARS) spaced every 30–40 kb, and markers to facilitate [...] selection”. Used as “landing pads” for integration of ~30kb-scale fragments, using CRISPR/Cas9 to cleave the integration site. The neochromosomes can contain entire metabolic pathways, either endogenous (after knocking-out the endogenous copies) or exogenous.