Difference between revisions of "Part:BBa K2145102"
 
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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: Forward, toward the clamps; Direction of RFP: Reverse, toward the clamps
 
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: Forward, toward the clamps; Direction of RFP: Reverse, toward the clamps
  
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This part is one of a 4-plasmid set (K2145100, K2145101, K2145102, and K2145103). Other plasmids in this set are similar, but have GFP and RFP expression cassettes pointing in different directions. 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. Addition of dCas and gRNAs matching the clamp sites are expected to reduce supercoiling, leading to more similar expression between the plasmids in this set. Addition of dCas and gRNAs matching the clamp sites are expected to reduce supercoiling, leading to more similar expression between the plasmids in this set.
+
This part is one of a 4-plasmid set (K2145100, K2145101, K2145102, and K2145103). Other plasmids in this set are similar, but have GFP and RFP expression cassettes pointing in different directions. 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. Addition of dCas and gRNAs matching the clamp sites are expected to reduce supercoiling, leading to more similar expression between the plasmids in this set.
  
 
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Latest revision as of 23:48, 30 October 2016


GFP/RFP reporter (Forward/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: Forward, toward the clamps; Direction of RFP: Reverse, toward the clamps

This part is one of a 4-plasmid set (K2145100, K2145101, K2145102, and K2145103). Other plasmids in this set are similar, but have GFP and RFP expression cassettes pointing in different directions. 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. Addition of dCas and gRNAs matching the clamp sites are expected to reduce supercoiling, leading to more similar expression between the plasmids in this set.

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]