Part:BBa_K4665001:Design

INPN

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 470
  Illegal PstI site found at 592
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 470
  Illegal PstI site found at 592
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 470
  Illegal PstI site found at 592
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 470
  Illegal PstI site found at 592
  Illegal NgoMIV site found at 54
  Illegal AgeI site found at 555
  
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

The INPN will be embedded in the outer cell membrane of E. coli. This can in turn be fused to a linker to present a protein on the outside of the bacterial cell.

Source

This ice nucleation protein is derived from Pseudomonas syringae. The codon optimized sequence for E. coli (BL21 DE3) has been derived from the research performed by Zhu et al. (2022).

References

Mergulhao, F.J.M. et al. (January 8, 2005). Recombinant protein secretion in Escherichia coli. Biotechnology Advances, 23(3): 177-202. https://doi.org/10.1016/j.biotechadv.2004.11.003

Zhu, Y., et al. (December 6, 2021). Surface display of carbonic anhydrase on Escherichia coli for CO2 capture and mineralisation. Synthetic and Systems biotechnology, 7(1): 460-473. https://doi.org/10.1016%2Fj.synbio.2021.11.008