Difference between revisions of "Part:BBa K3140005:Design"
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===Design Notes=== | ===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=== | ===Source=== | ||
− | Neurospora crassa | + | 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). | ||
+ | '''2.''' Zoltowski, B.D. ''et al.'' Conformational switching in the fungal light sensor Vivid. ''Science'' '''316''', 1054-7 (2007). |
Latest revision as of 10:57, 21 October 2019
VVD36-C73A - Truncated VIVID fluoroprotein derived from Neurospora crassa
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 434
- 1000COMPATIBLE 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).