Part:BBa_K1053208

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

HHR is expressed constantly and YF1 and FixJ is expressed too. In dark YF1 phospholylates FixJ response regulator and FixJ response regulator activates P_fixk2, so taRNA(taR12) is expressed. taR12 binds expressed HHR and inactivate HHR’s self cleavage. In blue light YF1 doesn’t phospholylate FixJ response regulator, so FixJ response regulator doesn't activate P_fixk2. taRNA doesn’t bind HHR and HHR does self cleavage and GFP’s RBS is exposed, so GFP is expressed.

Evaluation
The GFP fluorescence intensity was taken after main culture for 12 hours by Plate Reader in this evaluation method.

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

If HHR’s active site isn’t mutated difference between in light or dark is observed. This result is suggest that in light taRNA is expressed and inhibits HHR’s auto cleavage.RNA oscillator’s result is difference between taRNA’s existence or absence isn’t observed. We think that this difference is caused by difference between P_const.(low) and P_bad.

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
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 585
  Illegal NgoMIV site found at 657
  Illegal NgoMIV site found at 747
  Illegal NgoMIV site found at 765
  Illegal NgoMIV site found at 1251
  Illegal NgoMIV site found at 1544
  Illegal NgoMIV site found at 1638
  Illegal AgeI site found at 299
  Illegal AgeI site found at 1419
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 1308