Part:BBa_K3081056

sgRNA generator with promoter J23119

SgRNA Generator was designed base on the type II CRISPR/Cas9 system including T7 promotor, lacZa', crRNA(tracrRNA) and T7 terminator, with two BsaI cutting sites flanking lacZa' coding sequence. To facilitate the construction of sgRNA expression arrays for multiplex DnaA boxes, the Golden Gate Assembly method was adopted to insert the target-specific sequence between two BsaI cutting sites. For constant expression of sgRNA, they were placed under the control of J23119 promoter.

Figure1 Schematic cartoon of the DNA construct of BBa_K3081056

This part is used in our CRISPRri systemBBa_K3081058 to generate specific 20bp sgRNA binding to different DNA boxes on the OriC,the genome replication origin of E.coli. For more information, see BBa_K3081056

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 251
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 441
  Illegal BsaI.rc site found at 37