Coding

Part:BBa_K4437502:Design

Designed by: Marian Grell   Group: iGEM22_Calgary   (2022-10-12)
Revision as of 09:15, 12 October 2022 by Mariangrell (Talk | contribs) (→‎Design Notes)

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B0030-FLAG-phasin-HlyA (E. coli)


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

This sequence was designed for experiments comparing RBS strength and resulting FLAG-phasin-HlyA expression levels, with the purpose of improving the expression system to increase phasin yields. Specifically, the aim of this part was to increase, or improve, phasin expression as compared to the original BBa_K2260002 part. As such, this part was designed to produce the weakest expression levels of the four RBS-FLAG-phasin-HlyA parts in our part collection.

The restriction enzyme recognition sites for BstAPI and BstBI were also added on the end of the part, respectively, making the part compatible for digestion and ligation into the Tokyo Tech 2012 team’s BBa_K934001 plasmid for PHB expression.

Source

This part is based on the BBa_K2260002 part from the University of Calgary's 2017 iGEM team. The phasin (PhaP) gene is from R. eutropha, the HlyA tag is from E. coli, and the T7 promoter is from the T7 bacteriophage.

References