Part:BBa_K4179009

FcPT1 under Rhlr promoter + P2A + mCherry reporter

This composite part comprises of an umbelliferone-6 prenyl transferase FcPT1 (BBa_K4179002), under the rhlr-regulated promoter (BBa_R0071). Downstream to FcPT1 there is a P2A sequence (BBa_K4179005), which is a self-cleavage element that induces ribosomal skipping during translation of a protein inside the cell. Downstream to which there is an mCherry (BBa_K106005) reporter gene, and an rrnB terminator sequence (BBa_K4047025).

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal XbaI site found at 2218
  Illegal SpeI site found at 1006
  Illegal PstI site found at 1858
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 1423
  Illegal SpeI site found at 1006
  Illegal PstI site found at 1858
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal XbaI site found at 2218
  Illegal SpeI site found at 1006
  Illegal PstI site found at 1858
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal XbaI site found at 2218
  Illegal SpeI site found at 1006
  Illegal PstI site found at 1858
  Illegal NgoMIV site found at 200
  Illegal NgoMIV site found at 765
  Illegal AgeI site found at 1497
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI site found at 281
  Illegal SapI site found at 1077
  

Usage and Biology

In this genetic system, the mCherry expression is tied directly to the start codon of FcPT1, while not being covalently bound to it. In such a design the mCherry signal corresponds to a 1:1 ratio, at the very least, which allows for a lower estimate of FcPT1's expression.

FcPT1 enzyme is the first enzyme in the decursin biosynthesis pathway, which the team of Technion 2022 was attempting to clone into a bacterial system. For more information about the FcPT1 enzyme, visit its page in the registry (BBa_K4179002), or visit the team’s wiki page.

The team used this sequence in the rhlr-tdpp7-m/cherry plasmid, which already holds the rhlr gene.