Plasmid

Part:BBa_K4058031

Designed by: Sarah Cumberland   Group: iGEM21_Guelph   (2021-10-20)


CLE18 multiplex gRNA in AlcR-pHSN6A0

This plasmid contains a dCas9 region fused to a VP64 transcriptional activation domain. dCas9 is under ethanol inducible control by the AlcR promoter. This plasmid also contains a CLE18 multiplex gRNA which directs dCas9 to bind to five locations within CLE18's promoter.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 10436
    Illegal XbaI site found at 5564
    Illegal SpeI site found at 12599
    Illegal PstI site found at 4635
    Illegal PstI site found at 5929
    Illegal PstI site found at 6475
    Illegal PstI site found at 7897
    Illegal PstI site found at 8101
    Illegal PstI site found at 9343
    Illegal PstI site found at 15507
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 10436
    Illegal NheI site found at 7141
    Illegal SpeI site found at 12599
    Illegal PstI site found at 4635
    Illegal PstI site found at 5929
    Illegal PstI site found at 6475
    Illegal PstI site found at 7897
    Illegal PstI site found at 8101
    Illegal PstI site found at 9343
    Illegal PstI site found at 15507
    Illegal NotI site found at 5556
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 10436
    Illegal BglII site found at 2550
    Illegal BglII site found at 6731
    Illegal BglII site found at 7421
    Illegal BamHI site found at 1554
    Illegal BamHI site found at 2292
    Illegal BamHI site found at 5533
    Illegal BamHI site found at 7025
    Illegal BamHI site found at 9063
    Illegal BamHI site found at 10160
    Illegal XhoI site found at 5525
    Illegal XhoI site found at 6349
    Illegal XhoI site found at 7531
    Illegal XhoI site found at 8695
    Illegal XhoI site found at 11260
    Illegal XhoI site found at 12354
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 10436
    Illegal XbaI site found at 5564
    Illegal SpeI site found at 12599
    Illegal PstI site found at 4635
    Illegal PstI site found at 5929
    Illegal PstI site found at 6475
    Illegal PstI site found at 7897
    Illegal PstI site found at 8101
    Illegal PstI site found at 9343
    Illegal PstI site found at 15507
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 10436
    Illegal XbaI site found at 5564
    Illegal SpeI site found at 12599
    Illegal PstI site found at 4635
    Illegal PstI site found at 5929
    Illegal PstI site found at 6475
    Illegal PstI site found at 7897
    Illegal PstI site found at 8101
    Illegal PstI site found at 9343
    Illegal PstI site found at 15507
    Illegal NgoMIV site found at 4060
    Illegal NgoMIV site found at 7293
    Illegal NgoMIV site found at 13336
    Illegal AgeI site found at 1258
    Illegal AgeI site found at 1386
    Illegal AgeI site found at 1917
    Illegal AgeI site found at 11633
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 4349
    Illegal BsaI.rc site found at 3136
    Illegal SapI site found at 6267
    Illegal SapI.rc site found at 7734


[edit]
Categories
Parameters
None