Difference between revisions of "Part:BBa K3814051:Design"

 
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===Design Notes===
 
===Design Notes===
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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).
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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.
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Revision as of 15:38, 21 October 2021


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

n/a

References