Team:Michigan/Parts

Part Documentation

Switch 1.0 (adapted for thrombin) with additional cut sites and a GFP reporter was submitted as a part. This part has been demonstrated as an RNA toehold switch, which responds to a specific DNA trigger and expresses GFP, with low background, as seen in Figure 1 of the results (LINK) section. This provides a starting point for other teams looking to adapt toehold switches.

In order to ease the cloning process for future teams, an additional HindIII cut site was added between the switch and GFP. The HindIII cut site combined with the iGEM cut sites that surround the switch makes it easy to substitute the reporter.

Additionally, the part submitted gives the many other teams inspired by the same publications, an easy control for in vitro translation reactions. Since it already contains a full switch and reporter, it can be transformed and purified and then used directly with a cell-free expression system, without any further cloning. The trigger is short enough to be quickly and cheaply bought as a DNA oligo. Thus, future teams exploring cell-free expression systems can test their systems induced with a DNA oligo (positive reaction control) and without (negative reaction control).