Coding

Part:BBa_K4244051

Designed by: Arthur Vancolen   Group: iGEM22_Wageningen_UR   (2022-10-08)
Revision as of 15:26, 10 October 2022 by ArthurVancolen (Talk | contribs)


Type IC CRISPR Cas

The sequence includes two repetitive sequences recognisable by the Type-IC Cas proteins. The second repetitive sequence is modified to prevent recombination, so the spacer sequence that is placed in between does not get flipped out. The sequence found in the middle of the repeats is a random DNA sequence, that can be substituted by a spacer targeting a desired locus in the genome. BsaI sites can be found at the end of the first repeat and at the beginning of the second, so a new spacer can be integrated through simple restriction-ligation.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 2296
    Illegal XbaI site found at 537
    Illegal PstI site found at 1019
    Illegal PstI site found at 1249
    Illegal PstI site found at 2041
    Illegal PstI site found at 3314
    Illegal PstI site found at 3734
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 2296
    Illegal NheI site found at 13
    Illegal NheI site found at 291
    Illegal NheI site found at 4216
    Illegal PstI site found at 1019
    Illegal PstI site found at 1249
    Illegal PstI site found at 2041
    Illegal PstI site found at 3314
    Illegal PstI site found at 3734
    Illegal NotI site found at 1659
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 2296
    Illegal BglII site found at 3285
    Illegal XhoI site found at 2403
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 2296
    Illegal XbaI site found at 537
    Illegal PstI site found at 1019
    Illegal PstI site found at 1249
    Illegal PstI site found at 2041
    Illegal PstI site found at 3314
    Illegal PstI site found at 3734
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 2296
    Illegal XbaI site found at 537
    Illegal PstI site found at 1019
    Illegal PstI site found at 1249
    Illegal PstI site found at 2041
    Illegal PstI site found at 3314
    Illegal PstI site found at 3734
    Illegal NgoMIV site found at 694
    Illegal NgoMIV site found at 944
    Illegal NgoMIV site found at 1087
    Illegal NgoMIV site found at 1886
    Illegal NgoMIV site found at 2423
    Illegal NgoMIV site found at 2927
    Illegal NgoMIV site found at 4081
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None