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:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE 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.