Part:BBa_K5102003:Design
mTagBFP2
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 511
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 602
- 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 (ORFs), 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 in three ORFs, a single amino acid substitution, K16R, was introduced. This substitution was selected due to the biochemical similarity between lysine (K) and arginine (R), ensuring that the protein's functionality remained intact despite the change.
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.