Plasmid
pASapI

Part:BBa_K4006000

Designed by: Margaret Cook   Group: iGEM21_ASU   (2021-09-27)


pASapI

This plasmid is designed to integrate upstream and downstream of the photosystem II sequence (psbH) in the chloroplast of Chlamydomonas reinhardtii and was transformed into a mutant selection strain of C. reinhardtii, CC-4388 for our purposes. Selection strain C. reinhardtii is lacking photosystem II and cannot photosynthesize. Integrating pASapI rescues function, allowing the transformed algae to photosynthesize as a selective trait, bypassing the need for antibiotic resistance.

This plasmid backbone is readily engineered with the type IIS restriction enzyme, SapI.

Figure 1. Visualization of the plasmid backbone in benchling.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 2445
    Illegal XbaI site found at 3821
    Illegal PstI site found at 3833
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 4361
    Illegal NheI site found at 5790
    Illegal NheI site found at 6111
    Illegal PstI site found at 3833
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 3815
    Illegal XhoI site found at 3739
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 2445
    Illegal XbaI site found at 3821
    Illegal PstI site found at 3833
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 2445
    Illegal XbaI site found at 3821
    Illegal PstI site found at 3833
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1319
    Illegal BsaI site found at 3923
    Illegal BsaI.rc site found at 6517
    Illegal SapI.rc site found at 3800
[edit]
Categories
//cds
Parameters
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