Coding

Part:BBa_K1673205

Designed by: Jarrod Shilts   Group: iGEM15_Vanderbilt   (2015-09-17)

T7 Endonuclease (Homology Optimized)

Repair enzyme that induces double strand breaks at DNA base mismatches. Has homology with K1673204 minimized for including both in system.

Usage

Functionally, this gene is identical to the T7 endonuclease K1673204.(see the page for K1673204 for more information). This sequence has been modified to minimize the amount of homology with the original mutation-optimized T7 sequence. In total, our homology-minimization software reduced the amount of homologies greater than five bases by 79% compared to incorporating the same T7 sequence twice.

This decreased homology makes it possible to insert two copies of T7 endonuclease into the cell with a lower risk of recombination disrupting their expression. The redundancy of having multiple copies will then vastly reduce the chance of a complete loss of function for T7 activity, since both T7 loci would have to independently mutate in the same cell.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 4
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 66
    Illegal BsaI.rc site found at 193


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Categories
Parameters
None