Fig. 4. Artificial induction of semi-targeted RNA-independent transposition at preferred motifs.

(A) A region on pTarget exhibiting low integration activity (original, blue) was substituted with rationally engineered sequences (colored lines) based on TnsC- and TnsB-binding preferences, generating the indicated pTarget variants (pT-1 to pT6). (B) After performing biochemical transposition assays with the indicated pTarget substrates, integration reads were normalized and mapped to either the forward strand (fwd, red) or reverse strand (rev, black). The intended untargeted integration site based on optimized poly-A and TnsB consensus motifs is marked with a maroon triangle and dotted line; the representative region at right (850 to 900 bp) is shown to highlight consistency in integration events observed elsewhere on pTarget.