Part:BBa_K3453120

Rosewood DmMatK Trigger (Full) transcription unit

This part is a transcription unit of the Rosewood DmMatK Trigger (Full) (BBa_K3453020) for sequence-based detection of rosewood. The transcription is controlled by the T7 promoter (BBa_K2150031) and the strong SBa_000587 synthetic terminator (BBa_K3453000).

Usage and Biology

A toehold switch is an RNA–based device containing a ribosome binding site (RBS) and an ATG start codon embedded in a hairpin structure that blocks translation initiation [1]. The hairpin can be unfolded upon binding of a trigger RNA therefore exposing the RBS and the ATG start codon permitting translation of the reporter protein (Figure 1).

Figure 1. Toehold switches principle.

This part is a transcription unit of BBa_K3453020 which is a trigger for the 6 toehold switches we designed for sequence-based detection of rosewood (BBa_K3453011, BBa_K3453012, BBa_K3453013, BBa_K3453014, BBa_K3453015, BBa_K3453016).

This part proved to be functional: the transcribed rosewood RNA was able to bind to the corresponding toehold switches and a readable output was generated.

Full results are available on the BBa_K3453111, BBa_K3453112, BBa_K3453113, BBa_K3453114, BBa_K3453115, BBa_K3453116 pages in the registry.

References

[1] Green AA, Silver PA, Collins JJ, Yin P. Toehold switches: de-novo-designed regulators of gene expression. Cell (2014) 159, 925-939.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 78
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 78
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 78
  Illegal BamHI site found at 271
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 78
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 78
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 396