Plasmid

Part:BBa_K3775000:Design

Designed by: Tyler Myers   Group: iGEM21_SDSU   (2021-10-20)


M. maripaludis S2 sMMO integration Plasmid


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 6471
    Illegal EcoRI site found at 13597
    Illegal EcoRI site found at 16306
    Illegal XbaI site found at 3581
    Illegal SpeI site found at 16130
    Illegal SpeI site found at 16331
    Illegal PstI site found at 2431
    Illegal PstI site found at 4760
    Illegal PstI site found at 4811
    Illegal PstI site found at 5195
    Illegal PstI site found at 5285
    Illegal PstI site found at 5911
    Illegal PstI site found at 7243
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 6471
    Illegal EcoRI site found at 13597
    Illegal EcoRI site found at 16306
    Illegal SpeI site found at 16130
    Illegal SpeI site found at 16331
    Illegal PstI site found at 2431
    Illegal PstI site found at 4760
    Illegal PstI site found at 4811
    Illegal PstI site found at 5195
    Illegal PstI site found at 5285
    Illegal PstI site found at 5911
    Illegal PstI site found at 7243
    Illegal NotI site found at 9376
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 6471
    Illegal EcoRI site found at 13597
    Illegal EcoRI site found at 16306
    Illegal BglII site found at 2644
    Illegal BglII site found at 4906
    Illegal BglII site found at 5005
    Illegal BglII site found at 8633
    Illegal BglII site found at 15262
    Illegal BamHI site found at 5050
    Illegal BamHI site found at 8667
    Illegal BamHI site found at 16337
    Illegal XhoI site found at 4156
    Illegal XhoI site found at 4685
    Illegal XhoI site found at 5640
    Illegal XhoI site found at 7426
    Illegal XhoI site found at 16145
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 6471
    Illegal EcoRI site found at 13597
    Illegal EcoRI site found at 16306
    Illegal XbaI site found at 3581
    Illegal SpeI site found at 16130
    Illegal SpeI site found at 16331
    Illegal PstI site found at 2431
    Illegal PstI site found at 4760
    Illegal PstI site found at 4811
    Illegal PstI site found at 5195
    Illegal PstI site found at 5285
    Illegal PstI site found at 5911
    Illegal PstI site found at 7243
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 6471
    Illegal EcoRI site found at 13597
    Illegal EcoRI site found at 16306
    Illegal XbaI site found at 3581
    Illegal SpeI site found at 16130
    Illegal SpeI site found at 16331
    Illegal PstI site found at 2431
    Illegal PstI site found at 4760
    Illegal PstI site found at 4811
    Illegal PstI site found at 5195
    Illegal PstI site found at 5285
    Illegal PstI site found at 5911
    Illegal PstI site found at 7243
    Illegal NgoMIV site found at 1980
    Illegal NgoMIV site found at 3255
    Illegal NgoMIV site found at 9333
    Illegal NgoMIV site found at 10169
    Illegal NgoMIV site found at 10357
    Illegal NgoMIV site found at 10927
    Illegal NgoMIV site found at 11587
    Illegal AgeI site found at 9580
    Illegal AgeI site found at 13402
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 6714
    Illegal BsaI site found at 12620
    Illegal BsaI site found at 12685
    Illegal BsaI site found at 13799
    Illegal BsaI.rc site found at 1081
    Illegal BsaI.rc site found at 7642
    Illegal BsaI.rc site found at 7780
    Illegal BsaI.rc site found at 8197
    Illegal BsaI.rc site found at 9399
    Illegal BsaI.rc site found at 11786
    Illegal BsaI.rc site found at 12728
    Illegal SapI site found at 1920
    Illegal SapI site found at 2130
    Illegal SapI site found at 14075
    Illegal SapI site found at 14285


Design Notes

Operon structure, size, necessary genes for sMMO operation, and codon optimization in M. maripaludis S2 were all considerations for this part.


Source

The part originated from the plasmid pCRUptNeo, designed by the Leigh lab: https://faculty.washington.edu/leighj/markmutpro.html

References