Plasmid_Backbone

Part:BBa_K4188001:Design

Designed by: Tobias Bröker   Group: iGEM22_WWU_Muenster   (2022-09-27)


pSB3CY_aeBlue shuttle vector backbone for Saccharomyces cerevisiae


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 6155
    Illegal XbaI site found at 3425
    Illegal SpeI site found at 3790
    Illegal PstI site found at 1105
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 6155
    Illegal NheI site found at 3849
    Illegal NheI site found at 3872
    Illegal NheI site found at 4841
    Illegal SpeI site found at 3790
    Illegal PstI site found at 1105
    Illegal NotI site found at 4782
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 6155
    Illegal BamHI site found at 2441
  • 23
    INCOMPATIBLE WITH RFC[23]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 6155
    Illegal XbaI site found at 3425
    Illegal SpeI site found at 3790
    Illegal PstI site found at 1105
  • 25
    INCOMPATIBLE WITH RFC[25]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 6155
    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
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI site found at 1378
    Illegal BsaI site found at 6161
    Illegal BsaI.rc site found at 1099
    Illegal SapI site found at 1082
    Illegal SapI.rc site found at 6


Design Notes

During our project we faced difficulties for the transformation of our various constructs. With the given iGEM plasmids it was not possible for us to insert our gene cassettes as one of the four given places, inside the MTU, would always be occupied by a yeast specific selection marker. To circumvent this problem, we developed our own shuttle vector for yeast transformation. By inserting the selection marker on a distinct position, we were able to safe the full capacity of the MTU, further saving costs and time, by reducing the amount of needed transformations.


Source

This shuttle vector was assembled using synthesized parts, iGEM biobricks (BBa_K1430000,BBa_K864401, BBa_J23110, BBa_B0015, pSB3C01) and PCR fragments.

References