Composite

Part:BBa_K1604025

Designed by: Turcato Alessandro   Group: iGEM15_UNITN-Trento   (2015-09-12)
Revision as of 07:37, 16 September 2015 by Eli (Talk | contribs)

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. 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. PR creates a proton motive force across the membrane that can subsequently power cellular processes, such as ATP synthesis.


  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.

    3. 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


[edit]
Categories
Parameters
None