Coding

Part:BBa_K3140005:Design

Designed by: Fahad Ali   Group: iGEM19_Sydney_Australia   (2019-10-12)
Revision as of 10:49, 21 October 2019 by Fahad3li (Talk | contribs) (→‎References)


VVD36-C73A - Truncated VIVID fluoroprotein derived from Neurospora crassa


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 434
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This part was intended for use in pK18 as a test of our codon harmonisation model. o achieve this, the part was ordered in a gBlock flanked by EcoRI and HindIII sites to enable traditional restriction cloning into pK18. A RBS sequence was added to the gBlock upstream of the part. The RBS sequence used was the consensus Shine-Dalgarno sequence.

Source

The native sequence for VVD (Neurospora crassa) was obtained from NCBI (GenBank accession AF338412.1) and a coding sequence was generated by removing the first 36 amino acid residues and appending a start codon to the start of this truncated sequence. The cystine residue at position 73 (TGC) was then changed to alanine by changing the codon to GCC to yield VVD36-C73A.

References

1. Schwerdtfeger, C. & Linden, H. VIVID is a flavoprotein and serves as a fungal blue light photoreceptor for photoadaptation. EMBO J 22, 4846-55 (2003). 2. Zoltowski, B.D. et al. Conformational switching in the fungal light sensor Vivid. Science 316, 1054-7 (2007).