摘要：SummarySite-specific recombination systems (SRSs) are widely used in studies on synthetic biology and related disciplines. Nondirectional SRSs can randomly trigger excision, integration, reversal, and translocation, which are effective tools to achieve large-scale genome recombination. In this study, we designed 6 new nondirectional SRSs named Vika/voxsym1-4 and Dre/roxsym1-2. All 6 artificial nondirectional SRSs were able to generate random deletion and inversion inSaccharomyces cerevisiae. Moreover, all six SRSs were orthogonal to Cre/loxPsym. The pairwise orthogonal nondirected SRSs can simultaneously initiate large-scale and independent gene recombination in two different regions of the genome, which could not be accomplished using previous orthogonal systems. These SRSs were found to be robust while working in the cells at different growth stages, as well as in the different spatial structure of the chromosome. These artificial pairwise orthogonal nondirected SRSs offer newfound potential for site-specific recombination in synthetic biology.Graphical abstractDisplay OmittedHighlights•Designed six new artificial nondirectional site-specific recombination systems•Pairwise orthogonal nondirected recombination systems in yeast•The deletion efficiency of systems is far greater than the inversion efficiency•These nondirectional recombination systems were found to be robustBiological sciences; Biotechnology; Genetic engineering; Biological sciences research methodologies; Bioengineering; Synthetic biology
