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Part:BBa_K2066550:Design

Designed by: Kalen Clifton, Christine Gao, Andrew Halleran, Ethan Jones, Likhitha Kolla, Joseph Maniaci, John Marken, John Mitchell, Callan Monette, Adam Reiss   Group: iGEM16_William_and_Mary   (2016-10-14)


85x tetO Binding Array


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 2434
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 2434
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1593
    Illegal BamHI site found at 825
    Illegal BamHI site found at 959
    Illegal BamHI site found at 1109
    Illegal BamHI site found at 1726
    Illegal BamHI site found at 1934
    Illegal BamHI site found at 2515
    Illegal BamHI site found at 3081
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 2434
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 2434
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2893
    Illegal SapI.rc site found at 160
    Illegal SapI.rc site found at 2506


Design Notes

Cloned using restriction digest, as PCRs are incredibly difficult with this level of homology. While PCR is used to check for correct insert length in colonies, it often leads to multiple laddering off target bands.


Source

Plasmid and sequence obtained from addgene PRS303-240xtetO.


References

Finney-Manchester et al Nucleic Acids Res. 2013 Mar 7