Part:BBa_K1053206

Pconst.- RBS- YF1/FixJ-DT- Pfixk2- HHR*- GFP- DT

In dark, YF1 phospholylates FixJ response regulator and FixJ response regulator activates Pfixk2, so HHR* is expressed. HHR* contains GFP’s RBS. When HHR* is expressed, but this HHR is mutated in active site, so HHR doesn’t do self cleavage. GFP’s RBS isn’t exposed, so GFP isn’t expressed. This composition is used negative control.

Evaluation
The GFP fluorescence intensity was taken after main culture for 8 hours by Plate Reader in this evaluation method.

Result Evaluation of transformed E.coli under the light or dark condition.

There is no clear difference of light and dark condition of HHR-GFPuv or HHR*-GFPuv. The cause of this result is that excitation light of GFP and YF is almost the same, we think. Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 2815
  Illegal XhoI site found at 2716
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 605
  Illegal NgoMIV site found at 677
  Illegal NgoMIV site found at 767
  Illegal NgoMIV site found at 785
  Illegal NgoMIV site found at 1271
  Illegal NgoMIV site found at 1564
  Illegal NgoMIV site found at 1658
  Illegal AgeI site found at 319
  Illegal AgeI site found at 1439
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 1328
  Illegal BsaI.rc site found at 2230