Reporter
GG108

Part:BBa_K2145103

Designed by: Samuel Clamons, Katie Maruna, Laura Chen, Melody Wu   Group: iGEM16_Alverno_CA   (2016-10-14)


GFP/RFP reporter (Reverse/Reverse) separated by three dCas clamp sites

This plasmid contains a GFP and an RFP reporter separated by a 282-bp spacer containing three gRNA binding sites for dCas. The reporter is designed to demonstrate the effect, if any, of binding a dCas "clamp" between two genes (the idea being that such a clamp can limit propagation of supercoiling between the genes, functionally isolating them). Direction of GFP: Reverse, away from the clamps; Direction of RFP: Reverse, toward the clamps

This part is one of an 4-plasmid set (K2145100, K2145101, K2145102, K2145103). Other plasmids in this set are similar, but have GFP and RFP expression cassettes pointing in different directions and in different arrangements on the plasmid. Different arrangement and orientations are expected to generate different patterns of supercoiling, which translates to different patterns of expression in RFP and GFP if supercoiling creates interference between the two.

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 1411
    Illegal AgeI site found at 1523
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None