Part:BBa_K2483007

Ddx4-YFP

This is a CDS for a fusion proteins of Ddx4 and YFP. The N-terminal and C-terminal domain of Ddx4 have been shown to induce liquid-liquid phase separation in HeLa cells. There, the domains were fused to YFP and it could be seen under the fluorescence microscope that yellow fluorescing droplets would form in the cytoplasm.

Liquid-liquid phase separation occurs in all cells and can be imagined like oil droplets forming in water but here proteins are forming the droplets. We used this construct in yeast to show that it not only works in human cells and to see if we can use it for our metabolic channelling project (for that see BBa_K2483008: https://parts.igem.org/Part:BBa_K2483008 and BBa_K2483009: https://parts.igem.org/Part:BBa_K2483009).

A Kozak sequence was included to the part because in the plasmid we used it, there was no Kozak sequence included.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 790
  Illegal BamHI site found at 802
  Illegal BamHI site found at 1531
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 978
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 173
  Illegal SapI.rc site found at 832