Composite

Part:BBa_K1969007

Designed by: Yijie Lai   Group: iGEM16_SJTU-BioX-Shanghai   (2016-10-08)
Revision as of 13:45, 27 October 2016 by Charlotte Doyle (Talk | contribs)


Nano-lantern(cAMP-1.6) regulated by ADH1 promoter

The sequence of Nano-lantern(cAMP-1.6) is cloned downstream the yeast ADH1 promoter and ended with an ADH1 terminator to generate a expression cassette in yeast Saccharomyces cerevisiae.


Nano-lantern(cAMP-1.6) regulated by ADH1 promoter (BBa_K1969007)

Fig.1 The picture of mechanism. Upon binding with extracellular glucose, the Gpa2 protein will disassociate from Gpr1 and activates adenylyl cyclase, yielding cAMp surge which can be detected by our composite part product.


Nano-lantern(cAMP-1.6) contains a portion of enhance YFP with 10 amino acids deleted at C terminus, denoted as Venus△C10, a portion of mutated Renilla luciferase with 3 amino acids deleted at N terminus, Rluc8, and a cAMP binding domain of EPAC1 (Exchange Protein Directly Activated by cAMP) flanked by two separate parts of Rluc8. Upon binding with the cAMP molecule, the catalytic activity of the split luciferase will increase as the separate parts are brought together because of the conformational change in EPAC, thus results in the change in luminescence via the BRET effect. Thus this part can tell us the relative concentration of extracellular ligands.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 2101
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 90
    Illegal BsaI.rc site found at 1379
    Illegal BsaI.rc site found at 1963
    Illegal SapI.rc site found at 2164


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Parameters
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