Part:BBa_K612001

FadD 5

This is the 5'-UTR of the FadD gene of N. crassa. It is suitable for creating constructs which can knockout the FadD gene, as well knock in new genes.

We believe that this part potentially fulfills the silver medal requirement for iGEM. The parts plasmid it is contained in has been shown to:

• Transform properly into E. coli, and produce colonies on chloramphenicol plates

• Upon subsequent digestion of this parts plasmid with NotI and BsaI, it was shown that this part produces the exact correct bands. The parts plasmid used for this characterization was an adapted pSB1C3 with RFP inserted. As expected, we see a band at ~2000 bp (the plasmid backbone), and a band at ~850 bp (the part)

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• The parts plasmid, when digested with BsaI, produces the correct ~800 bp part. This part correctly ligates to parts with complimentary overhangs.

While the part itself is meant to work in Neurospora crassa, and the data for its use in N. crassa is still ongoing, we believe that in the spirit of the competition our entirely new part, designed to work with a brand new RFC (BBF RFC 82), deserves merit. Homologous regions such as this one are absolutely essential for work in Neurospora crassa, and the identification of our validated ends (through ligation) strongly contribute to the registry through BBF RFC 82. We believe that this part should be considered as a valuable and functionally validated contribution to the parts registry.

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
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 819