Plasmid

Part:BBa_K4190000

Designed by: N/A   Group: iGEM22_UCSC   (2022-09-19)


pET28:GFP

Vector for gene insertion and subsequent protein expression.

Usage and Biology

For the expression of the protein Exendin-4 in E. coli, we used the pET28:GFP [1] plasmid as our backbone. This plasmid has a prokaryotic origin of replication and kanamycin antibiotic resistance. We inserted our gene fragment into the space currently taken up by Green Fluorescent Protein (GFP).

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 192
    Illegal XbaI site found at 959
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 192
    Illegal NotI site found at 165
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 192
    Illegal BglII site found at 1025
    Illegal BamHI site found at 198
    Illegal XhoI site found at 158
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 192
    Illegal XbaI site found at 959
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 192
    Illegal XbaI site found at 959
    Illegal NgoMIV site found at 1057
    Illegal NgoMIV site found at 2645
    Illegal NgoMIV site found at 2805
    Illegal NgoMIV site found at 5852
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 3725


References

[1] D. L. Shis and M. R. Bennett, “Library of synthetic transcriptional AND gates built with split T7 RNA polymerase mutants,” Proc. Natl. Acad. Sci. U.S.A., vol. 110, no. 13, pp. 5028–5033, Mar. 2013, doi: 10.1073/pnas.1220157110.

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Categories
//plasmidbackbone
Parameters
None