Difference between revisions of "Part:BBa K3140005:Design"

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===Design Notes===
 
===Design Notes===
  
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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.
+
This part was based on the native VVD protein from (''Neurospora crassa''). The VVD36 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.
 +
 
 +
This part was intended for use in pK18 as a test of our codon harmonisation model. To 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===
 
===Source===
  
−
The native sequence for VVD (''Neurospora crassa'') was obtained from NCBI (GenBank accession [https://www.ncbi.nlm.nih.gov/nuccore/AF338412.1 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.
+
The native sequence for VVD (''Neurospora crassa'') was obtained from NCBI (GenBank accession [https://www.ncbi.nlm.nih.gov/nuccore/AF338412.1 AF338412.1]).
  
 
===References===
 
===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).
 
'''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).
 
'''2.''' Zoltowski, B.D. ''et al.'' Conformational switching in the fungal light sensor Vivid. ''Science'' '''316''', 1054-7 (2007).

Revision as of 10:56, 21 October 2019


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 based on the native VVD protein from (Neurospora crassa). The VVD36 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.

This part was intended for use in pK18 as a test of our codon harmonisation model. To 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).

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).