Difference between revisions of "Part:BBa K5102010:Design"
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===Source=== | ===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 sequence does not originate from a natural genomic sequence but was specifically engineered for this project. | + | 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=== | ===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 | 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 | ||
Latest revision as of 06:47, 30 September 2024
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- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 4
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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