Difference between revisions of "Part:BBa K5102002:Design"
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===Design Notes=== | ===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. | + | 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. Where possible, 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. To ensure the absence of STOP codons, an M22V mutation in mScarlet-3 was introduced, based on its homology to mRed7. Additionally, an N98V mutation was made, chosen due to the structural similarity of the amino acids involved, preserving the integrity of the protein. |
| − | + | ||
===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=== | ||
Latest revision as of 06:46, 30 September 2024
mScarlet3
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 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. Where possible, 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. To ensure the absence of STOP codons, an M22V mutation in mScarlet-3 was introduced, based on its homology to mRed7. Additionally, an N98V mutation was made, chosen due to the structural similarity of the amino acids involved, preserving the integrity of the protein.
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.