Part:BBa_K4239002

Enhanced luciferase subunits fiatluxD

Description

fiatluxD is to be used with fiatluxC (BBa_K4239001) and fiatluxE (BBa_K4239005). It codes for a subpart fatty acid reductase. With the subparts encoded by fiatluxC and fiatluxE, they form a complex that recycles fatty acids to fatty aldehydes. Fatty aldehydes will be used as a substrate for the luciferase protein.

The system fiatluxC/fiatluxD/fiatluxE is to be used with fiatluxA (BBa_K4239003) and fiatluxB (BBa_K4239004), gathered in the fiatluxCDABE (BBa_K4239008) operon.

fiatlux genes come from ilux genes (C, D, A, B, E). They were modified to remove every iGEM restriction site (EcoRI, XbaI, SpeI and PstI) 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 than the lux system.

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
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Construction

fiatluxD was directly synthesized with fiatluxC and the promoter J23117 (BBa_J23117). The description of the synthesis is on the following page part fiatluxCD (BBa_K4239006).

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.