Part:BBa_K4103001

pIP501 transfer origin

Origin of transfer to permit conjugation (of the entire pHT43 plasmid when assembled) to B. subtilis.

Biology

Origins of transfer are portions of genetic information necessary for conjugation. This sites usually named as oriT are cut prior to transfer.[1]

Usage

B. subtilis are not efficient strains for electroporation and other types of transformation[2], knowing this, our team designed this biobrick to allow the transfer of our plasmids from E. coli to our B. subtilis strains.

Name: pIP501 Transfer Origin BsaI Prefix: CGAA BsaI suffix: GCAA

This part contains the origin of transfer to permit conjugation (of the entire pHT43 plasmid when assembled) to B. subtilis. This part (pVA2241) was originally from the Streptococcal Plasmid pVA1702 (derivative of pIP501)

References Streptococcal Plasmid pIP501 Has a Functional oriT Site pVA2241, fig2 - https://www.ncbi.nlm.nih.gov/pmc/articles/PMC177163/pdf/1774199.pdf Designed plasmid by Keoni Gandall: pOpen1.4 -> contained this part. (POTENTIAL FOR A MUCH SMALLER oriT: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4981023/)"

Expressed in E. coli not Bacillus subtilis

References

[1]Low KB. Conjugation. Encyclopedia of Genetics. Published online 2001:449-453. doi:10.1006/rwgn.2001.0267 ‌

[2]Lu Y-P ., Zhang C, Lv FX, Bie XM, Lu Z-X . Study on the electro-transformation conditions of improving transformation efficiency for Bacillus subtilis. Letters in Applied Microbiology. 2012;55(1):9-14. doi:10.1111/j.1472-765x.2012.03249.x

‌ ‌ Sequence and Features

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]