Part:BBa_K3352008
T7 + New RBS & Terminator + SplintR Ligase Expressing Construct
Construct Design
We improved this construct by using pET3a vectors with appropriate BioBrick prefixes and suffixes that fulfill the assembly standard. pET vectors include the T7 bacteriophage gene 10, which promotes high-level transcription and translation. Utilizing both a T7 promoter, T7 terminator, and an extended UTR sequence around the RBS and before the terminator, we would maximize the protein expression for our enzymes.
Characterization
Our SDS-PAGE results show that both purified proteins migrate at the expected sizes. However, due to the presence of unfavorable buffer conditions in our eluted proteins, we performed a buffer exchange to reach the desired pH and storage conditions for our enzymes. We used these proteins for our viral detection test.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1029
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 869
- 1000COMPATIBLE WITH RFC[1000]
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