Impact of the stuffer properties on Mos1 transposition efficiency. Transposition assays in bacteria were performed using pKK-MOS1 as the Tpase source. (a) Transposition efficiency of Tet stuffer with various GC content. The Tet genes and the added fragments are shown as boxes; The GC contents of the Tet genes and/or added fragments are indicated in each box. ITRs are represented as arrows, flanking the stuffer. The total GC content of each stuffer is bracketed on the left. The transposition efficiency of each construct is indicated on the right. (b) Variation of transposition efficiency (vertical axis) with the GC content of stuffers (horizontal axis). The reference construct (3Tet3) is indicated. The axis of the ellipse follows the correlation between transposition frequencies and GC content of the stuffers. Only one construct (the last construct drawn in a.) does not fall within this ellipse. Averages and standard errors were calculated from at least five replicates.