Difference between revisions of "Part:BBa K1467100"

Revision as of 13:57, 24 September 2014

RFP coding device - Golden Gate Module Flipper

This part is a conversion of BBa_J04450 para "GoldenGate Flipper modules" that can be used to convert Pro + 5U (promoter and 5' untranslated leader) modules from the GoldenGate MoClo Assembly Standard into standard BioBricks. Flanking the RFP is an inverted pair of BsaI recognition sequences either side of the original RFP reporter part. The flipper reaction is a one-pot, one-step, digestion-ligation GoldenGate cloning reaction. The image below shows the sequences inserted (in orange) between the RFP and the BioBrick prefix and suffix that enable the flipper to accept MoClo Pro+5U parts, which begin GGAG and end AATG, in a one-step digestion-ligation Golden Gate cloning reaction.

The colonies containing this part are clearly red in color under natural light after about 18 hours. Smaller colonies are visibly red under UV. The RFP part does not contain a degradation tag and the RBS is strong.When cloning is successful the colonies become white as the RFP sequence is replaced by the new part. After the reaction is complete, no part of the BsaI recognition sequence will remain between the BioBrick suffix and the part.

Sequence and Features