title: Lambert Lab CRISPR-Cas12a Xseq Target-promoter/Intertarget Distance
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, there are 1 to 2 targets  downstream of a constitutive promoter, which are varied in position with  respect to each other or the promoter. The crRNA sequence is fixed. 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).