Signalling

Part:BBa_K608102

Designed by: Julia Müller   Group: iGEM11_Freiburg   (2011-09-14)

CcaS, green light receptor


CcaS is a cyanobacteriochrome, which is known to chromatically regulate the expression of the phycobilisome linker gene (cpcG2) in Synechocystis (see references). It functions as the receptor for green light and is made up of a N-terminal transmembrane helix, a cyanobactreria specific GAF domain, two PAS domains and a C-terminal histidine kinase. To be fully functional CcaS has to bind the chromophore Phycocyanobilin (PCB) with its GAF-domain. The GAF domain in this system has the ligation motif Cys-Leu, instead of the usual plant GAF-domain with Cys-His

FR2011 Green light system.jpg

Usage and Biology

The natural sequence has an internal EcoRI and SpeI restriction site.
We tried to delete these sites by PCR, but where not completely successful.
You can use this part in order to assemble the green light receptor system.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1685
    Illegal SpeI site found at 1780
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1685
    Illegal NheI site found at 1056
    Illegal SpeI site found at 1780
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1685
    Illegal XhoI site found at 6
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1685
    Illegal SpeI site found at 1780
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1685
    Illegal SpeI site found at 1780
    Illegal AgeI site found at 1720
  • 1000
    COMPATIBLE WITH RFC[1000]

Improvement

Designed by: Du liaoqi Group: iGEM18_WHU-China (2018-10-05)
We have condon-optimeized Ccas BBa_K2789009 for E.coli from BBa_K608102 and deleted the EcoRI and SpeI site in the gene sequence.
(https://parts.igem.org/Part:BBa_K2789001:Design)




References

Yuu Hirose et al ( 2008 )"Cyanobacteriochrome CcaS is the green light receptor that induces the expression of phycobilisome linker protein." Proc Natl Acad Sci U S A. 105: 9528–9533.

Tabor, J. J. et al.(2010), " Multichromatic Control of Gene Expression in Escherichia coli", J. Mol. Biol. , doi:10.1016/j.jmb.2010.10.038

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