title: Lambert Lab CRISPR-Cas12a Xseq Migrating Target
design: A compact synthetic dCas12a inverter is used to probe the binding strength of Cas12a to its DNA target. All components are contained within a plasmid, and the experiment is conducted in E. coli. Here, the target is a constitutive promoter. The crRNA and promoter sequences are varied nonrandomly in order to test the efficacy of binding to particular promoter regions. The constitutive promoter drives the production of a tetA/sacB cassette, which is selected for in the presence of tetracycline and selected against in the presence of sucrose. Cas12a binding is measured via selection with sucrose (SK), tetracycline (TK), or a kanamycin control (K).