Part:BBa_K4813005

J23100 - dTomato red chromoprotein strong expression construct

This composite component comprises several elements, including a constitutive strong promoter (J23100), a strong ribosome binding site (B0034), an E. coli codon-optimized chromoprotein and red fluorescent protein dTomato coding sequence (K4813000), and a strong double terminator (B0015). Additionally, the 5' and 3' ends of the composite part feature a 20 base pair overlap sequence designed for the pUC19 EcoRI restriction site for NEBuilder HiFi assembly.

Our project aims to create a formaldehyde sensing reporter. This composite part served three purposes:

Comparing expression levels of dTomato and tdTomato chromoproteins. Providing a reference for expression levels in our formaldehyde sensing reporter. Acting as a positive control to ensure successful cloning.

Literature suggests that tdTomato should have higher expression levels when expressed in E. coli colonies compared to dTomato. However, our results showed that E. coli colonies expressing dTomato had a brighter red color even under naked eyes. Therefore, our project proceeded with using dTomato constructs to increase the sensitivity of our formaldehyde reporter.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 27
  Illegal NheI site found at 50
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]