Coding

Part:BBa_K2657005

Designed by: Kim Ly   Group: iGEM18_Austin_UTexas   (2018-10-09)


Phytobrick of sYFP2 (mutational hotspot removed)

This part was improved from the BBa_K864100 Biobrick which encodes Super Yellow Fluorescent Protein 2


Originally, BBa_K864100 had an IS10 insertion sequence site, which resulted in an insertion in the region of: 5’ -acaggcgtagtaccg- 3’. In BBa_K2657005 (the improved part), the IS10 sequence has been removed and the area of the insertion sequence is now 5’ -actggggttgttcca- 3’. This sequence maintains the codon identity of the original sequence.


We then designed primers that added BsmBI and BsaI restriction sites to the sequence and inserted it into PhytoBrick universal acceptor, BBa_P10500, via BsmBI assembly. This created a Phytobrick that functions in MoClo Assemblies with BsaI, which is useful as a selectable marker due to its strong fluorescent character.




Figure 1. Colonies are non- fluorescent under blue light. White-circled colonies highlight what expected colonies look like with the PhytoBrick.







Sequence and Features


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]


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Categories
Parameters
None