Composite

Part:BBa_K5109023:Design

Designed by: Lisa Faccincani   Group: iGEM24_Uni-Padua-IT   (2024-09-28)
Revision as of 15:52, 29 September 2024 by Lisafaccincani (Talk | contribs) (References)

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Deha2 surface display system


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 639
    Illegal BamHI site found at 1372
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 667


Design Notes

We decided to exploit the first codon of the coding sequence to introduce NdeI cutting site, which is CATATG.

To better implement the design solution described, we modified the restriction enzyme sites compared to BBa K5109001: we removed the two BsaI cut sites downstream of the passenger, because at the same time, we introduced a BsaI restriction site upstream of the sequence encoding the passenger, allowing for more elegant manipulation of the surface expression cassette: with these improvements, it is now possible to change the passenger protein whether the carrier is present or removed from the expression cassette.



Source

This part is form from the assebly of different basic parts: it was designed on Benchling and subsequently synthesized.

References

iGEM20_USAFA work

Harris JD, Coon CM, Doherty ME, McHugh EA, Warner MC, Walters CL, Orahood OM, Loesch AE, Hatfield DC, Sitko JC, Almand EA, Steel JJ. Engineering and characterization of dehalogenase enzymes from Delftia acidovorans in bioremediation of perfluorinated compounds. Synth Syst Biotechnol. 2022 Feb 16;7(2):671-676. doi: 10.1016/j.synbio.2022.02.005. PMID: 35224235; PMCID: PMC8857417.

Hui CY, Guo Y, Liu L, Zheng HQ, Wu HM, Zhang LZ, Zhang W. Development of a novel bacterial surface display system using truncated OmpT as an anchoring motif. Biotechnol Lett. 2019 Jul;41(6-7):763-777. doi: 10.1007/s10529-019-02676-4. Epub 2019 Apr 25. PMID: 31025146.