Part:BBa_K4634016

supD

.We constructed an AND gate in E. coli that takes two stimulus signals from the environment as input, and the two signals activate the transcription of their respective regulatory genes. The AND gate can only be triggered when both regulatory genes are transcriptionally active and integrated with each other.

Integration occurs through interactions between mRNA and tRNA. The first regulatory gene is T7ptag (T7 RNA polymerase TAG); the second regulatory gene is tRNA supD, which is specifically used for the translation of T7ptag. When both regulatory genes are transcribed, T7ptag is translated and synthesized. T7ptag can specifically bind to the T7 promoter and translate the genes downstream of the T7 promoter (i.e., the genes regulated by the AND gate), thereby realizing the AND gate. Function[1].

As one of the elements that implements the AND gate, supD is a tRNA that recognizes TAG amber stop codons in T7ptag transcription product. Therefore these stop codons are translated as serine when supD is also transcribed, continuing the translation process of T7ptag that should have been terminated. As a result: when T7ptag and supD are transcribed at the same time, T7ptag can be translated and synthesized, and the genes downstream of the T7 promoter can be expressed (Fig. 1) [1].

Figure 1. A schematic representation of the genetic AND gate is shown.

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
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 27
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 30
  

References

[1] Anderson JC, Voigt CA, Arkin AP. Environmental signal integration by a modular AND gate. Mol Syst Biol. 2007;3:133. doi: 10.1038/msb4100173. Epub 2007 Aug 14. PMID: 17700541; PMCID: PMC1964800.