Extended Data Fig. 4: Strand Transfer Complex design and assembly.
From: Molecular basis for transposase activation by a dedicated AAA+ ATPase
a, The DNA substrate is composed of three different single-stranded DNA sequences. The longest molecule (130 nt) contains the complete sequence of the right TIR (orange) covalently bound to the target DNA (violet), including the insertion sequence (red). The second strand (60 nt) contains the complementary sequence of the right TIR (non-transferred strand, NTS), including a 5 nt-long 5’ overhang. The third strand (70 nt) contains the nucleotide sequence complementary to the target DNA. b, Step-wise formation of the substrate. Half-substrates are formed mixing and gently annealing the three DNA strands in equimolar amounts. The annealed substrate dimerizes via the insertion site aided by the proteins. c, Schematic of the final substrate displaying the complete sequence. d, Negative staining reference-free 2D averages show that the STC complex formed with wild-type IstB and the E167Q mutant are virtually identical at this resolution.