Fig. 3: Restructuring ChrI with CReATiNG.

a Chromosome I segments are cloned between a pair of position-specific adapters present into distinct versions of pASC1. Five possible restructured versions of BY ChrI were created by altering the adapters appended to each segment prior to their assembly. PCR of assembly junctions or ONT sequencing was used to confirm that the chromosome assemblies had the correct structure. b Growth analysis of the natural and five non-natural chromosome structures in a rich medium containing glucose at 30 °C. For each strain, a total of 12 independent growth experiments were carried out (n = 12). Each dot represents the value from an individual replicate of a given genotype, while the horizontal bar represents the mean across the 12 replicates. c The restructured ChrI containing the structures 1-3-2 and 3-1-2 present specific non-natural junctions involving the genes SNC1, MYO4, and NUP60. d Expression ratios of the genes SNC1, MYO4, and NUP60 in the control strain IEY402 (1-2-3) and restructured strains IEY421 (1-3-2) and IEY425 (3-1-2) were calculated using the Pfaffl method⁵⁰ and ACT1 as a control housekeeping gene. The values are the means and standard deviations of four independent biological replicates. Two-tailed t-tests with *, **, and *** corresponding to p < 0.05, p < 0.01, and p < 0.001, respectively. Exact p values are provided in Supplementary Table 15.