Plasmid

Part:BBa_K4188003

Designed by: Tobias Bröker   Group: iGEM22_WWU_Muenster   (2022-09-28)


pSB3CY_Leu shuttle vector with leucine synthase

This is the ready-to-use version of the pSB3CY_shuttlevektor backbone (BBa_K4188001) for Saccharomyces cerevisiae where the aeBlue chromoprotein is exchanged with the leucine synthase LEU2.It can be used for MoClo like the pSB3C01 backbone.


Usage and Biology

Test transformation

After a successful golden gate exchange of the aeBlue chromoprotein through the leucine synthase, the resulting vector pSB3CY_Leu was transformed into the yeast Saccharomyces cerevisiae.

Figure 1: left picture: Saccharomyces cerevisiae colonies formed from 100µl plated transformation mixture
right picture: Saccharomyces cerevisiae colonies formed from 10µl plated from the same transformation mixture
Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 7094
    Illegal XbaI site found at 3425
    Illegal SpeI site found at 3790
    Illegal PstI site found at 1105
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 7094
    Illegal NheI site found at 5780
    Illegal SpeI site found at 3790
    Illegal PstI site found at 1105
    Illegal NotI site found at 5721
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 7094
    Illegal BamHI site found at 2441
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 7094
    Illegal XbaI site found at 3425
    Illegal SpeI site found at 3790
    Illegal PstI site found at 1105
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 7094
    Illegal XbaI site found at 3425
    Illegal SpeI site found at 3790
    Illegal PstI site found at 1105
    Illegal AgeI site found at 793
    Illegal AgeI site found at 905
    Illegal AgeI site found at 2083
    Illegal AgeI site found at 2406
    Illegal AgeI site found at 4697
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1378
    Illegal BsaI site found at 7100
    Illegal BsaI.rc site found at 1099
    Illegal SapI site found at 1082
    Illegal SapI.rc site found at 6


[edit]
Categories
//chassis/eukaryote/yeast
//collections/shuttle_vectors/level2
Parameters
None