Coding

Part:BBa_K5527001:Design

Designed by: Otto Isaksson   Group: iGEM24_UppsalaUniversity   (2024-09-27)


amilCP purple, Purple chromoprotein


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

The substitution at position 157 in amilCP was induced through the use of inverse PCR with random primers specific to position 157. This is an area sequence-wise far away from the chromophore, but due to the folding of the protein it is located close enough to interact with it. To design this part, we transformed a pSB1K3 plasmid containing the green amilCP (BBa_K1996005, but without the T39S substitution from amilCP) into competent DH5-alpha E.coli. A high-copy plasmid is suitable for this purpose since it enables quicker and clearer screening of new colors. Inverse PCR was chosen since it avoids having to perform BioBrick assembly.



Source

amilCP provided by Anthony Forster at Uppsala University. amilCP originates from the coral Acropora millepora.

References