Part:BBa_K1065305

Blue light sensor without inverter for the production of amilGFP

This part is an improved version of the Blue light sensor device BBa_K952003 (see details in Design notes). In the presence of blue light (470 nm) the production of the reporter amilGFP is inhibited. In the absence of blue light the device is activated, thus producing amilGFP.
This part was cloned and successfully improved by UNITN-Trento 2013 iGEM team. We characterized the circuit with the control of pLac promoter (to do that we created the part BBa_K1065302) in order to test protein transcription and then add an ethylene forming enzyme (EFE) after amilGFP.

SAFETY NOTES: this part does not have safety concerns.

Usage and Biology

YF1, the blue light sensor, is a fusion protein of the LOV blue light sensor domain of Bacillus subtilis (YtvA) and FixL histidine kinase domain (from Bradyrhizobium japonicum). In the dark, the autophosphorylated YF1 phosphorylates FixJ, its Response Regulator, which activates the pFixK2 promoter allowing amilGFP transcription.
Under constant illumination with blue light net kinase activity is strongly suppressed, consisting in a consequent inactivation of pFixK2: amilGFP is no longer produced.
For characterization results please check BBa_K1065302 page. We inserted the pLac promoter upstream of the improved part whith RBS.

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
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 562
  Illegal NgoMIV site found at 634
  Illegal NgoMIV site found at 724
  Illegal NgoMIV site found at 742
  Illegal NgoMIV site found at 1254
  Illegal NgoMIV site found at 1547
  Illegal NgoMIV site found at 1641
  Illegal AgeI site found at 276
  Illegal AgeI site found at 1422
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 1311
  Illegal BsaI.rc site found at 175