Coding

Part:BBa_K5102010:Design

Designed by: Natalia Kuźmierkiewicz   Group: iGEM24_Munich   (2024-09-30)


eUnaG


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 4
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The design of the sequence was focused on optimizing codon usage for expression in HEK293T cells to ensure efficient protein expression. Additionally, care was taken to avoid introducing STOP codons in all three forward open reading frames, as their presence would disrupt our readout system. The sequence was also modified to be BioBrick compatible, allowing for easier modular assembly in synthetic biology applications. Importantly, these modifications were made without altering the amino acid sequence, preserving the functionality of the protein while enhancing its expression potential and compatibility with standard molecular biology techniques and our project.


Source

This part was ordered as a gBlock from a commercial DNA synthesis provider. It was designed synthetically with codon optimization for expression in HEK293T cells, ensuring compatibility with molecular biology techniques, including BioBrick assembly. The DNA sequence does not originate from a natural genomic sequence but was specifically engineered for this project.

References

Truong DJ, Geilenkeuser J, Wendel SV, Wilming JCH, Armbrust N, Binder EMH, Santl TH, Siebenhaar A, Gruber C, Phlairaharn T, Živanić M, Westmeyer GG. Exonuclease-enhanced prime editors. Nat Methods. 2024 Mar;21(3):455-464. doi: 10.1038/s41592-023-02162-w. Epub 2024 Feb 1. PMID: 38302659; PMCID: PMC10927552