Composite

Part:BBa_K1604010:Design

Designed by: Silvia Galvan   Group: iGEM15_UNITN-Trento   (2015-07-08)

araC-pBAD + Proteorhodopsin


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1740
    Illegal BamHI site found at 1144
    Illegal XhoI site found at 1313
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
    Illegal AgeI site found at 1502
    Illegal AgeI site found at 1877
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961


Design Notes

BBa_K773002 was extracted from the 2015 Registry distribution Kit. Proteorhodpsin was amplified by PCR with a designed forward primer containing a strong RBS. The new translation unit is BBa_K1604000 The amplified part was placed under an inducible promoter by Standard Assembly. Promoter used is arc-pBAD inducible by arabinose (BBa_K731201)


Source

BBa_K731201 + BBa_K1604000

araC-pBAD promoter comes from E.coli genome. Proteorhodopsin was taken from the uncultured bacterium HF10_19P19 of the SAR86 group of Gammaproteobacteria.

References

  1. Walter, Jessica M., Derek Greenfield, Carlos Bustamante, and Jan Liphardt. "Light-powering Escherichia Coli with Proteorhodopsin." Proceedings of the National Academy of Sciences 104 (2007): 2408-2412.

  2. Martinez, A., A. S. Bradley, J. R. Waldbauer, R. E. Summons, and E. F. Delong. "Proteorhodopsin Photosystem Gene Expression Enables Photophosphorylation in a Heterologous Host". Proceedings of the National Academy of Sciences 104.13 (2007): 5590-595.

  3. Richard A. Krebs, Ulrike Alexiev, Ranga Partha, Anne Marie DeVita, Mark S.Braiman. "Detection of fast light-activated H+ release and M intermediate formation from proteorhodopsin".BMC Physiology (2002), 1472-6793/2/5