Coding

Part:BBa_K3930003:Design

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


α-ionone induction system and expression in Saccharomyces cerevisiae (pViolette)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 5805
    Illegal PstI site found at 5798
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 5805
    Illegal PstI site found at 5798
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1265
    Illegal BglII site found at 1795
    Illegal BamHI site found at 1379
    Illegal BamHI site found at 3518
    Illegal BamHI site found at 4370
    Illegal XhoI site found at 73
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 5805
    Illegal PstI site found at 5798
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 5805
    Illegal PstI site found at 5798
    Illegal NgoMIV site found at 5303
    Illegal AgeI site found at 606
    Illegal AgeI site found at 5021
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 4761


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

NsrR, LcyE and ofCCD1 fragments were PCR amplified from IDT or Twist Bioscience gblocks and pCfB3040 XII-4 was linearized by inverse PCR.

References