Team:UCSF/Medal

Parts Submitted to the Registry

Construct containing YefM (anti-toxin) behind an IPTG inducible T7 promoter
- Part: BBa_K726012 - Link: BBa_K726012

Construct containing YoeB (toxin) behind an IPTG inducible T7 promoter
- Part: BBa_K726014 - Link: BBa_K726014

Construct containing three enzymes (VioA, VioB, VioE) behind an IPTG inducible T7 promoter
- Part: BBa_K726016 - Link: BBa_K726016

YoeB and YefM are new submissions to the registry and we have demonstrated that both constructs and parts work as desired.

Experience and Improvements with Existing BioBrick Parts

VioABE is an improvement on the previous submitted part by the 2009 Cambridge iGEM team (BBa_K274003). This is because our part contains only the three necessary genes to produce the green pigment. It is smaller and easier to clone, yet achieves the desired output.

The part necessary to produce the green pigment is now reduced in size from 5.9 kb to 4.8 kb
As reported by other teams in the Experience section, attempts to retrieve the Cambridge submitted part from the registry have failed. The restriction sites were not correct and we found that it failed as a PCR template for any of the four genes reported. Therefore, we obtained a template from another source (Dueber Lab, UC Berkeley) and are submitting the DNA for these three enzymes for use by future iGEM teams.

Finally, we have made the extraction of violacein (BBa_K274002) safer and more effective by showing that the pigment can be easily extracted with ethanol, rather than acetone or ethyl acetate, and that these samples are readily used in downstream analysis.