Part:BBa_K4239008

fiatlux genes with their promoter to emit luminescence

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 5342
  Illegal BamHI site found at 50
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 2993
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1366
  Illegal SapI site found at 3512
  

Description

The fiatlux operon is composed of 6 genes: fiatluxA, fiatluxB, fiatluxC, fiatluxD and fiatluxE, that form an autonomous system producing bioluminescence thanks to the encoded luciferase enzymes.

The genes fiatluxA and fiatluxB code each of them for a subunit of the luciferase protein. Both subunits need to be used together. Luciferase has as substrats FMNH2, O2 and Fatty aldehydes, and produces H20, Fatty Acids and FMN and emits luminescence.

The genes fiatluxC, fiatluxD and fiatluxE code each of them for a subpart of a fatty acid reductase. They need to be used together to form a complex that recycles fatty acids to fatty aldehydes. Fatty aldehydes will be used as a substrate for the luciferase protein.

Both systemes (fiatluxC/fiatluxD/fiatluxE and fiatluxA/fiatluxB) are made to be used together, and are gathered in the fiatluxCDABE operon.

Fiatlux genes come from ilux genes (C, D, A, B, E). They were modified to remove every Igem restriction site (EcoR1, Xba1, Spe1 and Pst1) included in genes. They were also adapted to include the biobrick format.

The ilux operon was born from a mutated natural luminescence operon present in the bacteria P.luminescens: the lux operon. These mutations were error-prone PCR induced according to Gregor et al.’s study in 2018 (Gregor et al. 2018). The aim was to create a system of genes that produced more light.

References

Gregor C, Gwosch KC, Sahl SJ, Hell SW. Strongly enhanced bacterial bioluminescence with the ilux operon for single-cell imaging. Proc Natl Acad Sci U S A. 2018 Jan 30;115(5):962-967. doi: 10.1073/pnas.1715946115. Epub 2018 Jan 16. PMID: 29339494; PMCID: PMC5798359.