Composite

Part:BBa_K1604025

Designed by: Turcato Alessandro   Group: iGEM15_UNITN-Trento   (2015-09-12)

araC-pBAD + proteorhodopsin + j23100 + blh + araC-pBAD + β-Carotene

This is the complete device for the use of proteorhodopsin, a light-powered proton pump that needs retinal as cofactor, without providing exogenous retinal.

Usage and Biology

This device contains the five genes necessary for retinal biosynthesis and the gene for the proteorhodopsin[1] [2] . In details it contains the four genes responsible of βcarotene production (ctrEIBY) under the control of araC-pBAD. The blh gene is under a costitutive promoter of the Anderson family (J23100) and encodes for the β-carotene 15-15 dioxygenase that catalyzes the cleavage of a single molecule of β-carotene into two molecules of retinal. Proteorhodopsin, taht is under the control of araC-pBAD, is a light-powered proton pump that needs retinal as cofactor. Proteorhodopsin creates a proton motive force across the membrane that can subsequently power cellular processes, such as ATP synthesis.

For more updates on the Unitn iGEM 2015 project check out our Wiki: Unitn iGEM wiki 2015


  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. Web.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 2019
    Illegal NheI site found at 2042
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1740
    Illegal BamHI site found at 1144
    Illegal BamHI site found at 4051
    Illegal BamHI site found at 6092
    Illegal XhoI site found at 1313
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 2257
    Illegal NgoMIV site found at 5628
    Illegal NgoMIV site found at 5758
    Illegal AgeI site found at 979
    Illegal AgeI site found at 1502
    Illegal AgeI site found at 1877
    Illegal AgeI site found at 3886
    Illegal AgeI site found at 4843
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961
    Illegal SapI site found at 3868


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Categories
Parameters
None