Part:BBa_K4274011

sfp_HA_US

sfp_HA_US is a homologous donor template situated in the upstream of sfp gene of Bacillus subtilis. After knock-out the natural sfp gene, sfp_HA_US could stimulate the homology-directed repair(HDR) of site-specific DNA double-strand breaks (DSBs) induced by the RNA-guided Cas9 endonuclease. Homology-directed repair (HDR) of these DSBs enables precise editing of the genome by introducing defined genomic changes, including base substitutions, sequence insertions, and deletions. It was used in the composite part PvanP*-sfp_target-sfp_HA_US-p43-K4274013-sfp-K4274014-degQ-sfp_HA_DS (Part: BBa_K4274035) to realize fengycins’ production in B. subtilis.

Source

Bacillus subtilis 168

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal XbaI site found at 1048
  
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal XbaI site found at 1048
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal XbaI site found at 1048
  
• 1000
  COMPATIBLE WITH RFC[1000]

References

[1]Nambiar T., Billon P., Diedenhofen G., et al. Stimulation of CRISPR-mediated homology-directed repair by an engineered RAD18 variant. Nature Communications. 10 (5), 2295 (2019). https://doi.org/10.1038/s41467-019-11105-z