Part:BBa_K4006007

IEE2

IEE2 was taken from the psbN-psbH intercistronic region, specifically. Macedo-Osoria et al. demonstrated its function by sucessfully expressing two proteins linked by IEE2 [1]. We attempted to utilize this intercistronic region to link two of the genes we hoped to express in C. reinhardtii, but ultimately were unable to clone the plasmid using this linker. IEE2 should only be utilized when the upstream and downstream flanking elements of the plasmid backbone are a significant distance from the the psbN-psbH junction. Should the IEE appear already in the plasmid backbone within the upstream or downstream flanking elements of a gene of interest, significant cloning errors may occur. For this reason, we failed to clone our combinations of genes (ArsC-Ftna, MT-FtnA, and MT-ArsC), since our plasmid backbone included the IEE2 sequence.

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
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]