Extended Data Fig. 2: Linear increase of k_on with number of repeats for 6 different sequence motifs.

Apparent \(k_{on}\) values were calculated by fitting the average dark times obtained from single binding sites on DNA origami with a Gaussian function and subsequently calculating \(k_{on} = 1/\left( {\tau _d \cdot c} \right)\), where c denotes the imager strand concentration. Error bars indicate standard deviation of Gaussian fits. All overlapping binding site motifs show a linear dependency until 5x repeats. Some 10x binding motifs exhibit a lower-than-expected \(k_{on}\), which could be explained by decreased accessibility due to the increased sequence length (and thus flexibility). Mean \(k_{on}\) values are additionally denoted in Supplementary Table 1. We note that each graph consists of data gathered from three separate experiments per sequence, containing repeat pairs of 1x and 3×, 1x and 5×, and 3x and 10×, respectively (see Supplementary Fig. 2). Furthermore, higher motif repeats were performed with reduced imager strand concentration to avoid multiple binding events. Plotting apparent k_on values allows us to faithfully compare sequence repeats (see Supplementary Tables 8 and 9 for details about image acquisition parameters and number of analyzed origami structures).