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Part:BBa_K2150027:Design

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


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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1561
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2858


Design Notes

Linker for fusion protein is carefully designed to avoid repeated sequence which may cause problems in assembly.

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

Fusion protein: 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).

pT7:PCR amplified from BBa_K525998

T7RNAP:PCR amplified from genome of BL21.

References