Regulatory
pT7-rbs

Part:BBa_K3114012:Design

Designed by: Cassandra Sillner, Sara Far, Sravya Kakumanu, Nimaya De Silva, Andrew Symes   Group: iGEM19_Calgary   (2019-10-08)


T7 inducible promoter and strong RBS


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1
    Illegal BsaI.rc site found at 67


Design Notes

When designing this part and the rest of our collection, we were interested in creating parts that could be used in Golden Gate assembly right out of the distribution kit without the need to first domesticate them in a Golden Gate entry vector. As such, these parts are not compatible with the iGEM Type IIS RFC[1000] assembly standard because we included the BsaI restriction site and MoClo standard fusion site in the part’s sequence.

As per the MoClo standard, the 5’ promoter fusion sequence included in this part is GGAG, and the 5’UTR 3’ fusion sequence is AATG (Weber et al., 2011).

Figure 1. Fusion sites used in the MoClo standard for Golden Gate assembly (Weber et al., 2011).

This part was designed in a manner that maintains the proper 8 bp AT-rich sequence between the RBS and the start codon, which is part of the 3’ fusion site.

Source

This part was synthesized.

References

Weber, E., Engler, C., Gruetzner, R., Werner, S., & Marillonnet, S. (2011). A modular cloning system for standardized assembly of multigene constructs. PLoS ONE, 6(2). https://doi.org/10.1371/journal.pone.0016765