Synthesis， debugging， and effects of synthetic chromosome consolidation: synVI and beyond.

We describe design， rapid assembly， and characterization of synthetic yeast Sc2.0 chromosome VI (synVI). A mitochondrial defect in the synVI strain mapped to synonymous coding changes within ()， encoding an essential proteasome subunit; Sc2.0 coding changes reduced Pre4 protein accumulation by half. Completing Sc2.0 specifies consolidation of 16 synthetic chromosomes into a single strain. We investigated phenotypic， transcriptional， and proteomewide consequences of Sc2.0 chromosome consolidation in poly-synthetic strains. Another "bug" was discovered through proteomic analysis， associated with alteration of the transcription start due to transfer RNA deletion and loxPsym site insertion. Despite extensive genetic alterations across 6% of the genome， no major global changes were detected in the poly-synthetic strain "omics" analyses. This work sets the stage for completion of a designer， synthetic eukaryotic genome.