Difference between revisions of "Part:BBa K4665005:Design"

(→‎Design Notes)
(→‎Design Notes)
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===Design Notes===
 
===Design Notes===
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This composite part results in a membrane fusion protein, meaning that extraction protocols for the protein should differ from normal extraction protocols
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The sequence of this composite part codes for the SazCA-INPN-Membrane module (SIMD). The resulting fusion protein will be embedded in the <i>E. coli</i> outer membrane by connecting the membrane-embedded N terminal of the Ice-Nucleation Protein () to the SazCA enzyme () via a flexible GGGGS linker ().
  
 
===Source===
 
===Source===

Revision as of 15:56, 8 October 2023


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 () to the SazCA enzyme () via a flexible GGGGS linker ().

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