Part:BBa_K3385075

PacI/Nt.BbvCI cassette

About the part: The part was obtained as part of the CRISPR cloning vector pFC330[1]. This is the PacI/Nt.BbvCI cassette which enables restriction enzyme digestion of the pFC330 plasmid to insert the CRISPR constructs.

Plasmid map of pFC330[1].

Functionality: After purification of the CRISPR cloning vector pFC330[1] in E. coli, this part enables linearization with the restriction enzyme PacI and the nicking enzyme Nt.BbvCI. Furthermore, USER overhangs are made in this process, which allows for USER assembly of biobricks.

Results: Purified plasmids were digested and run on a 1% agarose gel to verify linearization. The negative controls include non-linearized plasmid and MQ water.

PacI/Nt.BbvCI linearized plasmid pFC330[1] are viewed as clear single bands for 5 purified plasmids. Negative controls include: Non-linearized plasmid and MQ water.

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
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

References:
[1] A CRISPR-Cas9 System for Genetic Engineering of Filamentous Fungi. Nodvig CS, Nielsen JB, Kogle ME, Mortensen UH. PLoS One. 2015 Jul 15;10(7):e0133085. doi: 10.1371/journal.pone.0133085. eCollection 2015. PONE-D-15-11561 [pii] PubMed 26177455