Part:BBa_K4813014

Reverse HxlR-dTomato formaldehyde sensing chromoprotein reporter

This composite component comprises several elements, including BBa_K3332042 (composed of wild-type HxlR protein, a strong constitutive JS23100 reverse promoter, wild-type BRH1&2 binding sites for HxlR, a wild-type promoter called pFormaldehyde for Bacillus subtilis hxlAB operon and a wild-type Bacillus subtilis RBS site) from team iGEM20_XMU-China, an E. coli codon-optimized chromoprotein and red fluorescent protein dTomato coding sequence (BBa_K4813000), and a strong double terminator (BBa_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 (BBa_K4813007 & BBa_K4813008).

Fig. 1 Construct map showing the basic parts in this composite part.

The HxlR protein is a formaldehyde-responsive transcription factor from Bacillus subtilis [1]. In the presence of formaldehyde, wild-type HxlR protein binds to two specific binding sites (BRH1&2) and activates the expression of the downstream hxlAB operon in Bacillus subtilis [1]. In our project, we utilise HxlR to promote the expression of a red-colored dTomato chromoprotein when formaldehyde is present. In our project, this composite component serves the purpose of detecting the presence of formaldehyde by inducing a color change in the host E. coli.

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 442
  Illegal NheI site found at 465
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]