Publication date: 5 June 2020 http://www.sciencemag.org 2
Fig. 1. Characterization of CAST insertion products. (A) Schematic of genome targeting experiment
and summary of nanopore sequencing results. (B) Genetic assay for plasmid targeting. pInserts were
retransformed and selected on CmR+ and CmR+KanR+ plates to determine the fraction of co-integrate
insertions. The total insertion frequency was determined by droplet digital PCR and used to calculate the
co-integrate rate. (C) In vitro reactions with purified CAST proteins using plasmid donor or PCR-amplified
linear donor.