Part:BBa_K4212006

D15

Exonuclease coding sequence.

Usage and Biology

A level 0 golden gate reaction was set up to allow entry of the D15-1B sequence into the toolkit part collection. The assembly was then transformed in E. coli,plated onto LB Cm and colonies were screened via cPCR (Phire - Thermofisher).

Figure 1. The schematic design of D15

References

[1] Quijano, J.F. & Sahin, O. (2021) Genetically Intact Bioengineered Spores of Bacillus subtilis. ACS Synthetic Biology. 10 (4), 778–785. doi:10.1021/acssynbio.0c00578.

[2] Khromov, A.V., Makhotenko, A.V., Makarova, S.S., Suprunova, T.P., Kalinina, N.O. & Taliansky, M.E. (2020) Delivery of CRISPR/Cas9 Ribonucleoprotein Complex into Plant Apical Meristem Cells Leads to Large Deletions in an Editing Gene. Russian Journal of Bioorganic Chemistry. 46 (6), 1242–1249. doi:10.1134/S1068162020060138.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal XbaI site found at 52
  Illegal PstI site found at 812
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 812
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal XbaI site found at 52
  Illegal PstI site found at 812
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal XbaI site found at 52
  Illegal PstI site found at 812
  
• 1000
  COMPATIBLE WITH RFC[1000]