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Part:BBa_K3076500:Design

Designed by: Choi Jonas Sze Ngo   Group: iGEM19_Hong_Kong_JSS   (2019-10-05)


dsDNA substrate with KanR gene for cusF knockout in E. coli by Lambda Red Recombineering


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Source

The genomic sequence of cusF gene was extracted from NCBI (ACCESSION: NC_000913 REGION: 597131..597463)). The kanamycin resistance gene was extracted from the sequence of pET28a plasmid sequence from addgene.org. Double terminator was extracted from iGEM part registry (BBa B0015) which is the most commonly used double terminator with high efficiency.

Design Notes

The recombination site was chosen to be near the gene's 5' end of the coding strand to increase the chance of successful knockout.

References