RNA

Part:BBa_J100085:Design

Designed by: Caroline Vrana   Group: Campbell M Lab   (2012-07-13)
Revision as of 13:06, 23 July 2012 by Cavrana (Talk | contribs) (→‎Source)

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short CRISPR sequence with GFP target spacer


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal suffix found in sequence at 220
    Illegal XbaI site found at 15
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 221
    Illegal PstI site found at 235
    Illegal NotI site found at 6
    Illegal NotI site found at 228
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 214
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal suffix found in sequence at 221
    Illegal XbaI site found at 15
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 15
    Illegal SpeI site found at 221
    Illegal PstI site found at 235
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The CRISPR repeats made it difficult to not have inappropriate base pair matching with oligos.


Source

Oligos from a designed sequence. GFP target spacer came from part E0040. Leader sequence came from the 2011 USC iGEM team. The sequence for the CRISPR repeats comes from Touchon M, Rocha EPC (2010) The Small, Slow and Specialized CRISPR and Anti-CRISPR of Escherichia and Salmonella. PLoS ONE 5(6): e11126. doi:10.1371/journal.pone.0011126 and Mojica FJ et. al.Short motif sequences determine the targets of the prokaryotic CRISPR defence system. Microbiology. 2009 Mar;155(Pt 3):733-40.

References