Part:BBa_K4960024

pvc13 NTD-2*Bsal-pvc13 CTD

A sequence used to introduce the BsaI digestion site.

Usage and Biology

Introduce enzymatic cleavage sites required by the Golden Gate assembly method to test the effect of linkers with different structures on the structure of nonapeptide and tail filament protein. And then predicted the optimal display conditions for CKGGRAKDC by modeling. Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 309
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Special Design
From many different cloning strategies, we noticed one method named Golden Gate assembly, which achieves hierarchical assembly of DNA parts by utilizing Type IIS restriction enzymes to produce user-specified sticky ends on cut DNA fragments.[1] So we use Golden Gate Assembly to simplify the cloning process by altering the linker on both sides of the CKGGRAKDC to find the most suitable structure. This plasmid can be used as a tool plasmid for a series of subsequent plasmids.

Sequencing
This part is sequenced as correct after construction.
References
[1] Bird, J. E., Marles-Wright, J., & Giachino, A. (2022). A User's Guide to Golden Gate Cloning Methods and Standards. ACS synthetic biology, 11(11), 3551–3563.