This PDF file includes:

• fig. S1. The Isthmus of Panama in Tropical America.
• fig. S2. The current day Isthmus of Panama submerged under 150 m of relative sea-level rise.
• fig. S3. Timing of successful terrestrial dispersals between the American continents as observed in the occurrence of fossil remains in the rock record of North and South America.
• Legend for video S1
• table S1. Estimates of rates of uplift for the Panama Arc relative to sea level, using changes in estimated median depths and median ages of sedimentary units from previous studies (28, 135–149).
• table S2. Compilation of Late Paleocene–Late Eocene ages for the Colombian Andes plotted in Fig. 2.
• table S3. Median and 95% HPD intervals of the time at which members of clades spanning the Isthmus of Panama were separated from each other as estimated by BEAST (151) from phylogenies calibrated by fossils at one or more nodes.
• table S4. Kimura two-parameter distance between mitochondrial genes of sister clades on either side of the Isthmus of Panama.
• text S1. Estimation of date of splitting from molecular divergence.
• References (150–227)

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Other Supplementary Material for this manuscript includes the following:

• video S1 (.mov format). Large rafts form on the Río Chagres, Gamboa, Republic of Panama, on the 8th of December, 2010.