FIGURE 5.

Logo diagram of flanking extein sequences identified in the low stringency pilot study with the native Kan^R RBS reveals expanded specificity at the +2 and +3 positions. This figure was made using the Weblogo program (28) and shows 38 randomly chosen positive hits selected for splicing of the Npu DnaE intein in Kan^R site C. This construct results in expression of more Kan^R protein than needed for resistance and, therefore, allows selection of precursors that splice slowly or with a low percentage of spliced product (data not shown). The vertical red line indicates the position of the intein. The +1 position was fixed as Cys.