Part:BBa_K3183203

mClover3 with SpyTag and 6-His for E. coli

mClover3 is a 26.9 kDa protein derived from GFP. mClover3 improves photostability by 60% (t_1/2 = 80s) to its predecessor, owing to 2 mutations relative to dClover2: A206K and S160C. mClover3 can be used in Förster Resonance Energy Transfer (FRET) experiments with mRuby3, providing an alternative to cyan/yellow partners. This has the advantage of reducing spectral separation, having lower phototoxicity, and lower autofluorescence¹.

This construct was used for Spy&Go purification as a part of our characterisation of SpyTag, BBa_K1159201. Expression of this part was achieved in the pET28A vector BBa_K2141000.

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]

References

1) Bajar, Bryce T et al. “Improving brightness and photostability of green and red fluorescent proteins for live cell imaging and FRET reporting.” Scientific reports vol. 6 20889. 16 Feb. 2016, doi:10.1038/srep20889