Composite

Part:BBa_K4649009

Designed by: Sophia Eowyn Hsu   Group: iGEM23_EastCoastBioCrew   (2023-09-11)


pstSCAB codon optimized for R. tropici

pstSCAB is a high-affinity phosphate transporter protein. pstS is an inorganic phosphate (Pi) binding protein that delivers Pi to the membrane channel in the periplasm of a bacteria formed by pstA and pstC. pstB is an ATP hydrolysing protein that energizes the phosphate transport into the cell for the pstABC transporter. We used the lac promoter and lac operon in this part to increase its gene expression. Altogether, this operon increases the uptake of phosphate. We designed this part by codon optimizing the genes for rhizobia in Benchling. We obtained this DNA from IDT. We did not encounter any problems using this part. This part can be used in any biosafety level laboratory. This part is ideally used in rhizobia since it is codon optimized for that bacterium, but it can be adjusted for other chassises. To test this part, we ran a pcr with primers specific to this part, and ran a gel. Since the gel yielded positive results, we know that this part was successfully expressed. Therefore, this composite part is operational when codon optimized for rhizobia and our experiments suggest that it can be made compatible with rhizobia. During the gel electrophoresis we used a standard negative control to compare the results of the primers.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 2791
    Illegal XbaI site found at 3877
    Illegal SpeI site found at 3871
    Illegal PstI site found at 1667
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 2791
    Illegal SpeI site found at 3871
    Illegal PstI site found at 1667
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 2791
    Illegal BamHI site found at 1320
    Illegal BamHI site found at 3865
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 2791
    Illegal XbaI site found at 3877
    Illegal SpeI site found at 3871
    Illegal PstI site found at 1667
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 2791
    Illegal XbaI site found at 3877
    Illegal SpeI site found at 3871
    Illegal PstI site found at 1667
    Illegal NgoMIV site found at 207
    Illegal NgoMIV site found at 1248
    Illegal NgoMIV site found at 1430
    Illegal NgoMIV site found at 1490
    Illegal NgoMIV site found at 1988
    Illegal NgoMIV site found at 2823
    Illegal NgoMIV site found at 2912
    Illegal AgeI site found at 227
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1850
    Illegal SapI site found at 1369


[edit]
Categories
Parameters
None