Part:BBa_K656012

PowerCell

Anabaena vegetative psaC promoter (linked to photosystem I) with cscB sucrose symporter and GFPmut3b reporter.

This construct contains the coding sequence for cscB sucrose permease, a symporter which we use to secrete sucrose produced in our PowerCell organism, Anabaena 7120.

We place cscB under the control of psaC promoter, which we have shown to localize expression to vegetative Anabaena cells (as opposed to differentiated heterocysts, which are specialized nitrogen-fixing compartments that do not photosynthesize).

A GFPmut3b reporter was added to our construct to verify proper expression. The GFPmut3b variant has been previously shown to fluoresce effectively and be observed over background chlorophyll pigment levels in cyanobacteria (http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2860132/).

Transformed Anabaena carrying our full construct, Bright field image

Transformed Anabaena carrying our full construct, visualizing GFP reporter; Red arrows point to heterocysts

We have demonstrated that our construct is able to be taken up and properly expressed by Anabaena with our fluorescent reporter. Characterization of the sugar secretion function of cscB is underway and will be posted to this page as experiments are completed.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 697
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 697
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 697
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 697
  Illegal NgoMIV site found at 874
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 918
  Illegal BsaI.rc site found at 2319