Plasmid_Backbone
Gal4::EX-R

Part:BBa_K3271020:Design

Designed by: Setti Belhouari   Group: iGEM19_uOttawa   (2019-10-16)


Gal4::EX-RFP-SP pSB1K3 Type IIS Assembly


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found at 2290
    Illegal suffix found at 2346
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2290
    Illegal SpeI site found at 2347
    Illegal PstI site found at 2361
    Illegal NotI site found at 2296
    Illegal NotI site found at 2354
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2290
    Illegal XhoI site found at 1026
    Illegal XhoI site found at 2052
    Illegal XhoI site found at 2337
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 2290
    Illegal suffix found at 2347
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 2290
    Plasmid lacks a suffix.
    Illegal XbaI site found at 2305
    Illegal SpeI site found at 2347
    Illegal PstI site found at 2361
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI site found at 2312
    Illegal BsaI.rc site found at 2340


Design Notes

Please note the upstream and downstream Gal4 locus homologies are found outside the prefix and suffix, not within the multiple cloning site. This contribution consists of an updated pSB1K3 BioBrick backbone that makes the BioBrick plasmids compatible with cloning in both E. coli and S. cerevisiae. This plasmid backbone is compatible with traditional and Type IIS cloning.


Source

The Gal4 upstream and downstream homologies are comprised of 100 bp each from the promoter region and the terminator region of the Gal4 locus of S. cerevisiae, respectively. The RFP cassette is identical in sequence to BBa_J04450.

References