Part:BBa_K1467100
RFP coding device - Golden Gate Module Flipper
This part is a "GoldenGate Flipper" 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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 793
Illegal AgeI site found at 905 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1082
Illegal BsaI.rc site found at 6
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