Part:BBa_K2145000

GFP/RFP reporter (Forward/Forward) with random 500bp spacer

This part contains two fluorescent protein expression units (for RFP and GFP) with promoters, RBSs, CDSs, and terminators, both facing backward, with a random 500bp spacer between the two reporters. Parts were amplified from iGEM Registry (BBa_J04450 (RFP) and BBa_I13522 (GFP)) and assembled with golden gate assembly to make the full part. This composite part is designed to demonstrate the effect, if any, of a 500 bp spacer between genes on supercoiling generated by the two genes, and therefore on interference between the genes

This part is one of an 8-plasmid set (K2145000, K2145001, K2145104, K2145105, K2145124, K2145107, K2145126, and K2145127). 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 2226
  Illegal AgeI site found at 2338
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 724