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Part:BBa_K4968006:Design

Designed by: Shouye Zhu   Group: iGEM23_XJTLU-CHINA   (2023-10-09)


RecomSwapNeo R/Kan R


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 67
    Illegal XbaI site found at 1459
    Illegal PstI site found at 636
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 636
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 423
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 67
    Illegal XbaI site found at 1459
    Illegal PstI site found at 636
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 67
    Illegal XbaI site found at 1459
    Illegal PstI site found at 636
    Illegal NgoMIV site found at 1087
    Illegal NgoMIV site found at 1370
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 936
    Illegal SapI.rc site found at 1146


Design Notes

Considering that we need to knock out the CsgA and CsgB genes in the genomes of BL21(DE3) and MC4100 by ourselves. By searching in NCBI, we designed homologous fragments of 50 bp upstream and downstream of CsgA and CsgB genes. And the kanamycin sequence was referenced from pKD4. On the new plasmid, we designed 50 bp homologous fragments upstream and downstream of the kanamycin gene.

Source

the RecomSwapNeo R/Kan R fragments are from pKD4. The homologous fragments up and downstream of the CsgA and CsgB in the genome are from NCBI.