Coding

Part:BBa_K4437503:Design

Designed by: Marian Grell   Group: iGEM22_Calgary   (2022-10-12)


B0034-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 as a positive control for experiments comparing RBS strength and resulting FLAG-phasin-HlyA expression levels, with the purpose of improving the expression system to increase phasin yields. As such, this part was designed with the same RBS type as the original BBa_K2260002, to compare the relative expression levels of the other variable RBS types when combined with the FLAG-phasin-HlyA sequence.

The restriction enzyme cut sites for BstAPI and BstBI were also added, 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