Part:BBa_K4438605

T6_Trigger_3_w/oPol

T6_Trigger_3_w/oPol is single-stranded DNA having 29 nucleotides. Figure 1B)Illustrates the secondary structure and its minimum free energy. The 3’ end has ssDNA sense T7 promoter sequence.

Usage and Biology

Few bases of this part are complementary (Watson-Crick base pairing) to T6 (BBa_K4438600) and are designed to block its sequence motif. Motif, mostly associated with the loops, is linked to the formation of the specific binding site for testosterone [1]. Figure 2D) Shows the secondary structure of both parts hybridised at 37° Celsius. The hormone binds with T6 (BBa_K4438600) with high affinity and displaces the T6_Trigger_3_phi29. This part has complete complementarity with part T6_Binder_3(BBa_K4438602). T7 polymerase binds and in-vitro transcription of the duplex forms multiple broccoli light-up aptamers. Different levels of testosterone, an anabolic androgenic steroid (AAS), in biological fluids can be detected using all these three parts.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal SpeI site found at 4
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal SpeI site found at 4
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal SpeI site found at 4
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal SpeI site found at 4
  
• 1000
  COMPATIBLE WITH RFC[1000]

References

Skouridou, V., Jauset-Rubio, M., Ballester, P., Bashammakh, A. S., El-Shahawi, M. S., Alyoubi, A. O., & O’Sullivan, C. K. (2017). Selection and characterization of DNA aptamers against the steroid testosterone. Microchimica Acta, 184(6), 1631-1639.