Part:BBa_K5109023:Design
Deha2 surface display system
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 639
Illegal BamHI site found at 1372 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE 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.