Part:BBa_K1943020

R-GECO

GECO family is a series of fluorescent proteins that artificially engineered to response to changes in the concentration of calcium ion Ca2⁺. They can help to visualize intracellular signaling activity by the intensity. According to Zhao’s article, there is optimally 110-folds of emission change.

By recombining the GCaMPs and GFP, firstly they got the G-GECO1(green). The indicator we used is R-GECO1 derived from mApple and G-GECO1.1. The results of direct evolution also included GEM-GECO and Blue GECO. (see the genealogy of GECO series )

These various indicators is helpful in live cell image, and improve Ca2+ imagining in neurons and muscles as well. In our experiment, R-GECO1 was used to detect cell response to mechanical vibration.

There have been an iGEM team submitted its sequence in 2012 (BBa_K881000). But, unfortunately, it did not meet the RFC10 standard because of the restriction sites in its original sequence. For the convenience, we mutated these regions to remove the repeated cloning sites.

Mutated R-GECO CDS without 2 PstI in original sequence.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 914
  Illegal SapI.rc site found at 111
  Illegal SapI.rc site found at 820