Translational_Unit

Part:BBa_K4275030:Design

Designed by: Wei Mingshan   Group: iGEM22_GreatBay_SCIE   (2022-10-09)


pRha-riboj


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 91
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 91
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 91
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 91
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 91
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

1. The pRha promoter is a rhamnose-inducible promoter in which the transcription activity of the downstream CDS is tightly regulated by the concentration of rhamnose in the medium.

2. The 75bp ribozyme transccript forms stem-loop secondary structures, which cleave the upstream RNA leader and form a short hairpin structure. The CDS of RFP is 25bp downtream of the RiboJ DNA seuqence.

3. This RFP expression construct is utilized in the magnetic-recycling experiment of ferritin-expressing E.coli (BBa_K4275031 & BBa_K1189025). The red fluorescence of RFP facilitates the observation of the movement of magnetic field-sensitive E.coli towards the poles of the magnet situated in the centre of the petri dish.

Source

Synthetic construct
tobacco ringspot virus (sTRSV)

References

1.Clifton, Kalen P et al. “The genetic insulator RiboJ increases expression of insulated genes.” Journal of biological engineering vol. 12 23. 29 Oct. 2018, doi:10.1186/s13036-018-0115-6