Part:BBa_K2722002:Design

6 times Chi recombination hotspot to inhibit Exonuclease V (RecBCD) activity

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]

Design Notes

The sequence was ordered as 6 separate Primers with length between 33 and 39 bases. From these 6 primers we created 3 Primer-pairs complementing each other with a 4 base- overhang to the next primer-pair and create. These dimers were ligated together into the Backbone pSB1C3.

Source

Sequence was obtained from Marshall et al., 2017 (5) and ordered from IDT.

References

(1) Singleton, M. R., Dillingham, M. S., Gaudier, M., Kowalczykowski, S. C., & Wigley, D. B. (2004). Crystal structure of RecBCD enzyme reveals a machine for processing DNA breaks. Nature, 432(2014), 187.

(2) Smith, G. R. (2012). How RecBCD enzyme and Chi promote DNA break repair and recombination: a molecular biologist's view. Microbiology and Molecular Biology Reviews, 76(2), 217-228.

(3) Smith, G. R. (2001). Homologous recombination near and far from DNA breaks: alternative roles and contrasting views. Annual review of genetics, 35(1), 243-274.

(4) Dillingham, M. S., & Kowalczykowski, S. C. (2008). RecBCD enzyme and the repair of double-stranded DNA breaks. Microbiology and Molecular Biology Reviews, 72(4), 642-671.

(5) Marshall, R., Maxwell, C. S., Collins, S. P., Beisel, C. L., & Noireaux, V. (2017). Short DNA containing χ sites enhances DNA stability and gene expression in E. coli cell‐free transcription–translation systems. Biotechnology and bioengineering, 114(9), 2137-2141.

UniProt: 1w36 - RecBCD/DNA-complex