Reporter

Part:BBa_K1467400

Designed by: Cara Deal, Jessica Gray   Group: iGEM14_NRP-UEA-Norwich   (2014-09-24)
Revision as of 15:00, 24 September 2014 by Nicolapatron (Talk | contribs)

RFP coding device - Golden Gate Module Flipper

This part is a "GoldenGate Flipper" that can be used to assemble complete transcriptional units from GoldenGate MoClo Standard Parts producing a standard BioBrick. It consists of the RFP reporter (from BBa_J04450) flanked by an inverted pair of BsaI recognition sequences. The image below shows the sequences inserted (in orange) between the RFP and the BioBrick prefix and suffix that enable the flipper to accept the parallel assembly of multiple MoClo parts in a one-step digestion-ligation Golden Gate cloning reaction.

TUflipper.jpg

The colonies containing the RFP 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 the flipper reaction is successful the colonies become white as the RFP sequence is replaced by the new part. No part of the BsaI recognition sequence will remain between the BioBrick suffix and the new part.

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
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 793
    Illegal AgeI site found at 905
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1082
    Illegal BsaI.rc site found at 6


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