Reporter

Part:BBa_K2805007:Design

Designed by: Cassandra Scott Leach   Group: iGEM18_BostonU   (2018-10-10)


pGal1(Zif268op)x6-mRuby2


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 411
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 411
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal suffix found in sequence at 1608
    Illegal EcoRI site found at 411
    Illegal BglII site found at 891
    Illegal BamHI site found at 477
    Illegal XhoI site found at 885
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 411
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 411
    Illegal NgoMIV site found at 1928
    Illegal AgeI site found at 501
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The primary concern was Dueber MoClo and GoldenGate compatibility. These sequences were designed to be robustly assembled and integrated into the S. cerevisiae genome.

Source

Meghan Bragdon and Nikit Patel of the Khalil Lab at Boston University designed pGal1(Zif268op)x6 by mutating the pGal1 promoter and inserting six Zif268 binding sites.

References

1. Benzinger D, Khammash M. Pulsatile inputs achieve tunable attenuation of gene expression variability and graded multi-gene regulation. Nature Communications. 2018;9(1):3521. https://doi.org/10.1038/s41467-018-05882-2. doi: 10.1038/s41467-018-05882-2.

2. Hochrein L, Machens F, Messerschmidt K, Mueller-Roeber B. PhiReX: A programmable and red light-regulated protein expression switch for yeast. Nucleic Acids Res. 2017;45(15):9193-9205. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5587811/. doi: 10.1093/nar/gkx610.

3. Lee ME, DeLoache WC, Cervantes B, Dueber JE. A highly characterized yeast toolkit for modular, multipart assembly. ACS Synth Biol. 2015;4(9):975-986. https://doi.org/10.1021/sb500366v. doi: 10.1021/sb500366v.