Plasmid_Backbone

Part:BBa_K5194000:Design

Designed by: Nirmal Cheema   Group: iGEM24_UCalgary   (2024-09-17)


pSB1K0 Yeast Shuttle Vector (sfGFP dropout) Type IIS


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 41
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 41
    Illegal NheI site found at 1371
    Illegal NheI site found at 2498
    Illegal NheI site found at 2521
    Illegal NotI site found at 3502
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 41
    Illegal BglII site found at 154
    Illegal BamHI site found at 69
    Illegal XhoI site found at 4555
  • 23
    INCOMPATIBLE WITH RFC[23]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 41
  • 25
    INCOMPATIBLE WITH RFC[25]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 41
    Illegal NgoMIV site found at 4968
    Illegal NgoMIV site found at 5094
    Illegal NgoMIV site found at 5104
    Illegal AgeI site found at 231
    Illegal AgeI site found at 5350
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI site found at 3436
    Illegal BsaI.rc site found at 2466
    Illegal SapI site found at 4625
    Illegal SapI site found at 4996
    Illegal SapI.rc site found at 2577


Design Notes

The backbone was designed to be compatible with the open yeast collection for both level 1 and level 2 assemblies. It includes cut sites around the E. coli selection marker, allowing it to be swapped out, which makes the backbone distinguishable between level 1 and level 2 assemblies. The plasmid has a GFP dropout, which allows for visualization of successful assemblies.

Source

The genomic sequence comes from Komagataella pastoris and Pseudomonas aeruginosa.

References

The design was inspired by the JUMP vector collection. Marcos Valenzuela-Ortega, Christopher French, Joint universal modular plasmids (JUMP): a flexible vector platform for synthetic biology, Synthetic Biology, Volume 6, Issue 1, 2021, ysab003, https://doi.org/10.1093/synbio/ysab003