Part:BBa_K4438809

MN4_trigger_5_5�trigger_phi29

MN4_trigger_5_5’trigger_phi29(BBa_K4438809) is single-stranded DNA having 10 nucleotides. Figure 1B) shows the secondary structure and minimum free energy. The sequence is complementary to the 5’ region of the Aptamer_MN4 (BBa_K4438800) with no mismatched bases.

Usage and Biology

The part MN4_trigger_5_5’trigger_phi29(BBa_K4438809) has complementarity to the part Aptamer_MN4 (BBa_K4438800) and disrupts its three-way junction. Figure 1D) Shows the secondary structure of both parts hybridised at 37° Celsius. Quinine binds with cocaine-binding aptamer MN4 [1] and displaces the MN4_trigger_4_phi29(BBa_K4438807). This part has complete complementarity with MN4_Target_3(BBa_K4438806). Phi 29 DNA extension polymerase extends the template strand and in-vitro transcription of the duplex forms multiple broccoli light-up aptamers. Different levels of quinine can be detected using all these three parts.

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
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

References

Reinstein, O., Yoo, M., Han, C., Palmo, T., Beckham, S. A., Wilce, M. C., & Johnson, P. E. (2013). Quinine binding by the cocaine-binding aptamer. Thermodynamic and hydrodynamic analysis of high-affinity binding of an off-target ligand. Biochemistry, 52(48), 8652-8662.