Coding

Part:BBa_K3930001:Design

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


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


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 7298
    Illegal SpeI site found at 7208
    Illegal PstI site found at 7201
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 7298
    Illegal NheI site found at 4342
    Illegal SpeI site found at 7208
    Illegal PstI site found at 7201
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 7298
    Illegal BglII site found at 3135
    Illegal BglII site found at 3318
    Illegal BamHI site found at 753
    Illegal BamHI site found at 3483
    Illegal XhoI site found at 710
    Illegal XhoI site found at 7605
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 7298
    Illegal SpeI site found at 7208
    Illegal PstI site found at 7201
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 7298
    Illegal SpeI site found at 7208
    Illegal PstI site found at 7201
    Illegal NgoMIV site found at 6797
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 6646
    Illegal SapI.rc site found at 6856


Design Notes

According to Takara kit handbook advice and our experiments, a GC-rich homology zone is less likely to work for inFusion cloning. We therefore advise future iGEM teams to have homology zones for InFusion that flank such sequences


Source

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

References