Part:BBa_K3885353

P28-tetO-degfp-MGapt

Under the P28 promoter, the Tet operator tetO is expressed along with the production of green fluorescent protein.The transcription rate of the P28 promoter can be detected in response to MGapt.

Usage and Biology

Malachite green aptamer (MGA), an RNA aptamer that can specifically bind to malachite green (MG) dye and light up the fluorescence more than 2000-fold compared to negligible fluorescence of free MG. The MGA-MG system can be used for the quantitative analysis of RNA.
In Cell-Free systems, the strength of the P28 promoter has not been quantified. We cloned the MG aptamer on the 3’ end of P28-tetO-deGFP. Therefore, the transcription rate of P28 can be estimated. The optimal Ex/Em wavelengths are 625/655nm. The dye should be used under 20 μM, otherwise it is toxic to the reaction.
Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 763
  Illegal BamHI site found at 795
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]