Device

Part:BBa_K2428001:Design

Designed by: Patricia Zulueta   Group: iGEM17_UChicago   (2017-10-26)


Yeast-Bacteria shuttle vector


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1602
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1767
    Illegal XhoI site found at 3568
    Illegal XhoI site found at 4460
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 820
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1004
    Illegal SapI.rc site found at 448


Design Notes

We wanted the yeast selection marker (which makes the plasmid a shuttle vector) to be located within the backbone of the plasmid. In other words, so that other teams could easily put genes into the shuttle vector plasmid, we did not want the yeast selection marker to be within the prefix/suffix regions of psB1C3. This led us to design psB1C3mut, which has blunt-end restriction sites for cloning not within the prefix/suffix region of psB1C3. This is why we put the SCARG4 and PARS2 genes within psB1C3mut.



Source

The lab we work in provided us with a plasmid, pOW1, which contains SCARG4 (a gene that produces arginine) and PARS2 (an autonomous replicating element).

References