Composite
RCA system

Part:BBa_K5291030:Design

Designed by: Rui Chen   Group: iGEM24_BNUZH-China   (2024-09-27)


pBBRMCS2-acaP


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 242
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 242
    Illegal NotI site found at 845
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 171
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 242
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 242
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We introduce acaP into Rhodopseudomonas palustris in the form of multiple copies in order to accelerate the intake of CO2 released by Pseudomonas aeruginosa during the degradation of PE microplastics.
The gene acaP is from Rhodopseudomonas palustris CGA009, which ensures its expression in our engineered R. palustris. And the terminator BiTerm and the T7 promoter can be universally used in the bacteria we choose so that the composite part is able to come into effect normally.

Source

acaP: Rhodopseudomonas palustris CGA009
BiTerm: Escherichia coli
T7 promoter: T7 bacteriophage

References

[1]Puskas, L.G., et al., A periplasmic, alpha-type carbonic anhydrase from Rhodopseudomonas palustris is essential for bicarbonate uptake. Microbiology (Reading), 2000. 146 ( Pt 11): p. 2957-2966.
[2]Thorbecke, R., et al., The gene dosage effect of carbonic anhydrase on the biosynthesis of poly ( 3-hydroxybutyrate ) under autotrophic and mixotrophic culture conditions. POLYMER JOURNAL, 2021. 53(1): p. 209-213.
[3]Chen YJ, Liu P, Nielsen AA, Brophy JA, Clancy K, Peterson T, Voigt CA. Characterization of 582 natural and synthetic terminators and quantification of their design constraints. Nat Methods. 2013 Jul;10(7):659-64. doi: 10.1038/nmeth.2515. Epub 2013 Jun 2. PMID: 23727987.