DNA

Part:BBa_K3697003:Design

Designed by: Christopher Neimeth   Group: iGEM20_Stanford   (2020-10-22)
Revision as of 23:36, 23 October 2020 by Cneimeth (Talk | contribs) (→‎Source)


Homology Arms for KanR integration in B. Subtilis


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

Two major design considerations were made during the designing of this sequence. One, we had to make sure that each of the homology arms were at least 500 bp as they had to be of sufficient length to allow a recombination event in B. subtilis to occur. Two, we wanted to make sure that the sequence was something that would be easy for us to acquire and make, so we chose a portion of the pOpen yeast plasmid as this was something we had a lot of in the lab we were working with.


Source

These homology arms originally come from the pOpen Yeast plasmid and contain the portion of this plasmid that codes for the Kanamycin resistance gene. The design for this system were inspired by the work done by Josef Altenbuchner and Marian Wenzel [2]

[2] Wenzel, M. and Altenbuchner, J. (2015) Development of a markerless gene deletion system for Bacillus subtilis based on the mannose phosphoenolpyruvate‐dependent phosphotransferase system. Microbiology (United Kingdom), 161(10), 1942–1949.

References