Coding

Part:BBa_K2739004:Design

Designed by: Lok In Lo   Group: iGEM18_Edinburgh_OG   (2018-10-08)


Phasin


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
    COMPATIBLE WITH RFC[1000]


Design Notes

Available Phasin biobrick (BBa_K208001) found in Lac promoter-Phasin-HlyA biobrick, BBa_K390501. Through sequencing, it was found to contain a stop codon. A set of PCR primers was designed and used to remove the stop codon. In the primer design, a HindIII restriction enzyme site, which is not present in the original biobrick, was introduced and allow the confirmation of the success stop codon removal.

T--Edinburgh_OG_BBa_K2739007--image_5.jpeg Figure 1. The sequence alignment of BBa_K390501, LacP-Phasin-Hly-VR-170918-08-27(The sequencing result of BBa K2739007) and original-phasinHlyA-VR-170918-08-28 (The sequencing result of BBa_K390501) via Benchling. The selected (blue) part of the sequence indicated the stop codon.

Source

The phasin basic part and sequence information were obtained from the Composite part of BBa_K390501. (And Phasin sequence optimised by previous team)

References

1. Yamada M, Takahashi S, Okahata Y, Doi Y, Numata K. (2013 ) Monitoring and kinetic analysis of the molecular interactions by which a repressor protein, PhaR, binds to target DNAs and poly[(R)-3-hydroxybutyrate]. AMB Express. 3(1):6.