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Other
Part:BBa_K4968006:Design
Designed by: Shouye Zhu Group: iGEM23_XJTLU-CHINA (2023-10-09)
RecomSwapNeo R/Kan R
Sequence and Features
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 67
Illegal XbaI site found at 1459
Illegal PstI site found at 636 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 636
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 423
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 67
Illegal XbaI site found at 1459
Illegal PstI site found at 636 - 25INCOMPATIBLE 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 - 1000INCOMPATIBLE 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.