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Part:BBa_K2150026:Design

Designed by: Jianyi Huang   Group: iGEM16_UCAS   (2016-10-12)


pT7+pTet+RBS+tetX-GFP(fusion protein)+RBS+T7RNAP+DT


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 670
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1967


Design Notes

PT7 is placed upstream of pTet, regarding that pT7 is a strong promoter while pTet is relatively weak. Linker for fusion protein is carefully selected to avoid repeated sequence that may cause problems in assembly.


Source

pT7:PCR amplified from BBa_K525998;T7RNAP:PCR amplified from genome of BL21.

As for fusion protein tetX-GFP, tetX is the same as (xxx), the protein linker is provided by our advisor Li Hua, and the GFP is from BBa_E0040. We combine this three parts using overlap extension polymerase chain reaction (OE-PCR).

pTet from BBa_R0040, DT from BBa_B0015,RBS from BBa_B0034


References