Other

Part:BBa_K3814051:Design

Designed by: Simon Tang   Group: iGEM21_Sydney_Australia   (2021-10-01)


KpnI site


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]


Design Notes

The USYD 2021 team aimed to create naturally transformable (NT) bacteria by inserting the genes involved for NT from A. baylyi into a strain of E. coli. Due to the large number of genes (23) the genes were inserted sequentially in gene clusters. These were designed and created on SnapGene® software (from Insightful Science; available at snapgene.com).

One of the standard practices adhered to in the creation of these was that each cluster began with a KpnI restriction enzyme site, had a HindIII site between the last A. baylyi gene and the homology arm for the next cluster, and finally a BamHI site between the selectable marker and the terminator before the 3’ homology arm. This would allow for any necessary cuts/breaks to be made in the cluster, if say the cluster wasn’t working optimally and experimental diagnostics would need to be conducted to determine the problem with it.


Source

USYD iGEM 2021