Composite

Part:BBa_K3352009

Designed by: Hannah Hsu   Group: iGEM20_TAS_Taipei   (2020-10-18)
Revision as of 16:21, 18 October 2020 by Registry (Talk | contribs)

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T7 + New RBS & Terminator + Phi29 DNA Polymerase Expressing Construct

We improved this construct by using pET11a 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. 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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 676
    Illegal SapI.rc site found at 1074


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