Coding

Part:BBa_K3930002:Design

Designed by: Thomas Gaudin   Group: iGEM21_Toulouse_INSA-UPS   (2021-10-07)


β-ionone induction system and expression in S. cerevisiae (pFRAMBOISE-notfused)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 7891
    Illegal SpeI site found at 7801
    Illegal PstI site found at 7794
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 7891
    Illegal SpeI site found at 7801
    Illegal PstI site found at 7794
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 7891
    Illegal BamHI site found at 753
    Illegal BamHI site found at 4082
    Illegal XhoI site found at 710
    Illegal XhoI site found at 8198
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 7891
    Illegal SpeI site found at 7801
    Illegal PstI site found at 7794
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 7891
    Illegal SpeI site found at 7801
    Illegal PstI site found at 7794
    Illegal NgoMIV site found at 7390
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2988
    Illegal SapI.rc site found at 7239
    Illegal SapI.rc site found at 7449


Design Notes

According to our advisors and the Takara kit handbook advice, we separated the cloning in two steps for an optimum of efficacy


Source

Promoter Teto7 was PCR amplified from the addgene plasmid #165976, the NeoR fragment was PCR amplified from IDT gblock, the CrtY and fyn-phCCD1 were amplified from Twist Bioscience gblocks while pCfB3034 X-3 was linearized by inverse PCR on Addgene template plasmid.

References