Figure 1.

Schematic of Tn7 transposon mutagenesis library and insertion identification in S. uvarum. (A) Simplified representation of in vitro transposition of the Tn7 transposon into a plasmid library containing random S. uvarum genomic DNA fragments. Approximately 50,000 plasmids containing the Tn7 transposon were pooled together to form the final library. (B) Illustration of the Tn7-containing excised portion of the plasmid integrated into haploid and diploid yeast through homologous recombination. Approximately 500,000 NatR clones of each ploidy were pooled into two separate pools (haploid pool and diploid pool). (C) Design of Tn7-seq libraries used to identify insertion sites through sequencing. Reads containing Tn7 sequence are enriched (PCR off common flanking region of the Tn7 and Illumina adapter sequences) and mapped to the genome to identify insertion sites.