Composite

Part:BBa_K3352009:Design

Designed by: Hannah Hsu   Group: iGEM20_TAS_Taipei   (2020-10-18)


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


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


Design Notes

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.


Source

We obtained the amino acid sequence of phi29 DNA polymerase from Bacillus phage phi29.

References