Device

Part:BBa_K4461505:Design

Designed by: SHIH-HSUN, YANG   Group: iGEM22_NYCU-Taipei   (2022-10-10)


glpABC+RBS+(mCherry+tag)+BBa_B0015


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 993
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 993
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 993
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 993
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 993
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We cloned the promoter from E.coli K-12 MG1655 by PCR. Then we used PCR again to add suffix and prefix. Also, we separated the plasmid that we want to insert the promoter into two parts. Therefore, we can use Gibson Assembly to ligate three fragments. Then, we mutated the stop codon on mCherry and added EcoRI and HindIII cutting site by PCR twice. At last, we can insert the commercial tag by enzyme digestion and ligation.


Source

gene synthesis

References