FIGURE 1.

Placement of hairpin inserts in two color fluorescence vector. (A) Map showing placement of RFP and EGFP with respective CMV promoters. SacI and BamHI restriction sites encompass transcriptional start site for EGFP. The arrow indicates the relative location of the transcription start site. (B) Sample folding of the −10, −25, and −50 kcal/mol hairpin structures. A total of seven hairpins were utilized including −10, −20, −25, −30, −35, −40, and −50. (C) Example of the −30 kcal/mol hairpin placed at positions +1, +7, and +13. An insert length of 70 nucleotides was maintained with CAA repeats. Different hairpins were placed at positions +1, +4, +7, +10, +13, +16, +31, and +46. A complete listing of hairpins inserted into vectors GR1 and GR2 is shown in Supplemental Figure S1. Thermal stability predictions and hairpin structures were drawn with RNAstructure 3.7 (Mathews et al. 1999).