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[Preprint]. 2023 Jul 15:2023.07.14.549009. [Version 1] doi: 10.1101/2023.07.14.549009

Figure 1.

Figure 1.

DNA sequence specificity of GAF-DBD is kinetically defined. a, Schematics of 2-color smFRET experiment to measure cognate motif binding of GAF-DBD. b, Single-molecule trajectories showing Cy3-labeled GAF-DBD binding to Cy5-labeled DNA containing a GAF cognate motif. c, Representative single-molecule trajectories of Cy5-cognate DNA bound by Cy3-GAF-DBD showing an initial donor-only period before acceptor signal increases. The donor-only period is highlighted in grey. d, Zoomed-in view of binding events boxed in b. “*” indicates transient donor-only fluorescence within a binding event. e, Single-molecule trajectories showing Cy3-labeled GAF-DBD nonspecifically binding to Cy5-labeled non-cognate DNA. f, Binding frequency of GAF-DBD to cognate or non-cognate DNA. g, Dwell time of GAF-DBD on cognate or non-cognate DNA from fitting 1-CDF to an exponential decay function. Error bars show standard deviation of three technical replicates.