Part:BBa_K1969007

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.

This composite part consists of the full length yeast ADH1 promoter BBa_K1969003, the cAMP reporter protein Nano-lantern (cAMP1.6) BBa_K1969000, the terminator BBa_K1486025 and two artificially designed scars, BBa_K1969008 and BBa_K1969009.

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