Part:BBa_K2669000

Strongly constitutive His-tagged+flexible linker UnaG

WORK IN PROGRESS

UnaG is a unique chromoprotein since it expresses fluorescent signal when in contact with bilirubin. This means (unlike most other chromoproteins) it can be used as a reporter in anaerobic environments or even potentially in environments where bilirubin is naturally present, such as the intestines. While researching the previous work from the iGEM Uppsala 2016 team, we noticed that their part (Part:BBa_K2003011) had an error in it (a misplaced start codon) which would lead to not only reduced UnaG expression but also not include the histidine tag their part should be expressing according to the igem registry!

We decided that we would rectify this issue by engineering a composite part with the start codon in the correct position and compare it to the previous part (Part:BBa_K2003011). We designed this composite part by combining a strong promoter (Part:BBa_J23119), a strong RBS (Part:BBa_J34801), and a double terminator (Part:BBa_B0014) around the modified UnaG part (Part:BBa_K2669001).

WarningThis part also contains a GSG linker that has a stop codon directly after it. If you would like to use Part:BBa_K2669001 as a linker protein, you must delete or move this stop codon.

Source

As mentioned above, this part contains a strong promoter (BBa_J23119), a strong RBS (BBa_J34801), and a double terminator (BBa_B0014). In addition, it contains the sequence from BBa_K2003011, only with a change to the start codon location, resulting in Part:BBa_K2669001. The original source of UnaG is from the paper “A Bilirubin-Inducible Fluorescent Protein from Eel Muscle” by Kumagai A et. al, which characterized the UnaG protein from the muscle of a species of Japanese eel.

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
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]