Part:BBa_K4960024

pvc13 NTD-2*Bsal-pvc13 CTD A sequence used to introduce the BsaI digestion site.

Profile

Name: pvc13_NTD-2*Bsal-pvc13_CTD
Base Pairs: 1326bp
Origin: Synthetic
Properties: 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.

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.