Part:BBa_K4621083

CRISPRi-441

This CRISPRi fragment contains the dCas9 and sgRNA necessary for the CRISPRi system. The CRISPRi system is suitable for a variety of Streptomyces.[1] In this study, it was used to inhibit the expression of gabT gene in SCUT-3 to produce more GABA.

Usage and Biology

Due to the limited types of plasmids available for SCUT-3, we inserted the CRISPRi-239 fragment into the previously constructed plasmid by homologous recombination to achieve simultaneous stable expression of multiple target genes.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal SpeI site found at 4530
  Illegal PstI site found at 2424
  Illegal PstI site found at 2658
  Illegal PstI site found at 3870
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 4507
  Illegal SpeI site found at 4530
  Illegal PstI site found at 2424
  Illegal PstI site found at 2658
  Illegal PstI site found at 3870
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 463
  Illegal BglII site found at 1258
  Illegal BglII site found at 4027
  Illegal BamHI site found at 203
  Illegal BamHI site found at 1552
  Illegal BamHI site found at 4495
  Illegal BamHI site found at 4700
  Illegal XhoI site found at 876
  Illegal XhoI site found at 3222
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal SpeI site found at 4530
  Illegal PstI site found at 2424
  Illegal PstI site found at 2658
  Illegal PstI site found at 3870
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal SpeI site found at 4530
  Illegal PstI site found at 2424
  Illegal PstI site found at 2658
  Illegal PstI site found at 3870
  Illegal NgoMIV site found at 1820
  Illegal NgoMIV site found at 2191
  Illegal NgoMIV site found at 2394
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 3402
  Illegal BsaI.rc site found at 4281
  Illegal SapI site found at 794
  Illegal SapI site found at 2084
  Illegal SapI.rc site found at 3281