attTn7 target activity. (A) The E. coli attTn7 sequence from +23 to +59 is shown; nucleotides that are highly conserved and subsequently analyzed by mutation are boxed. (B) Graphical representation of the relative target DNA binding activity of TnsD on the mutant attTn7 site in vitro is shown. Double stranded oligonucleotides with indicated mutations listed were used in gel-shift assays with TnsD. (C) Graphical representation of the relative in vitro transposition activity on the mutant attTn7 site in vitro is shown. The activity of mutant attTn7 containing plasmid as a target for attTn7 transposition was measured using the lambda hop assay. The activity of wt attTn7 is 2 × 10-4 and the activity of mutants were normalized to this value.