Part:BBa_K4665005:Design

SazCA-INPN Membrane Display Module

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 sequence of this composite part codes for the SazCA-INPN-Membrane module (SIMD). The resulting fusion protein will be embedded in the E. coli outer membrane by connecting the membrane-embedded N terminal of the Ice-Nucleation Protein (BBa_K4665001) to the SazCA enzyme (BBa_K4665120) via a flexible GGGGS linker (BBa_K4665175).

Source

SazCA is derived from Sulfurihydrogenibium azorense and INPN is derived from Pseudomonas syringae. Both sequences have been codon optimized for E. coli (BL21 DE3).

Our sequence for SazCa is derived from the paper by Zhu et al. (2022), which has been codon-optimized for E. coli BL21(DE3).

References

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