DNA

Part:BBa_K4438807

Designed by: Shreyas Samir Parkhie   Group: iGEM22_IISER-Tirupati_India   (2022-10-12)
Revision as of 13:01, 12 October 2022 by Shreyasparkhie17 (Talk | contribs)

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MN4_trigger_4_phi29

MN4_trigger_4_phi29(BBa_K4438807) is single-stranded DNA having 28 nucleotides. Figure 1B) shows the secondary structure and minimum free energy. The middle region of the part is the ssDNA antisense T7 promoter sequence. Both ends have a few bases complementary to Aptamer_MN4 (BBa_K4438800).


Usage and Biology

The part MN4_trigger_4_phi29(BBa_K4438807) 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.

T--IISER-Tirupati India--mn4structure4.png 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.

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