BBa_K1616014 1 VVD VVD - homodimer photoreceptor 2015-09-17T11:00:00Z 2015-09-18T02:54:35Z VVD was isolated from Neurospora crassa. Among other photoreceptors, Vivid (VVD) is the smallest known Light???oxygen???voltage (LOV) domain protein and photo-inducible dimer. Isolated from Neurospora crassa, VVD forms a homo-dimer in response to a blue-light stimulus. The LOV domain, present in VVD, is a small blue-light sensing domain found in prokaryotes, fungi and plants. After blue-light activation, a covalent bond is formed between the co-factor Flavin mononucleotide (FMN) and one of the cysteine residue. This bond leads to a conformational change inducing functions by dissociating the C-terminal a-helix (Ja) and the LOV-core. In VVD, this undock triggers homodimerization (Bilwes, Dunlap, & Crane, 2007; M??ller & Weber, 2013). Contrary to other photoreceptors, VVD is a small protein with 150 amino-acids facilitating accurate molecular design and avoiding steric issues. Moreover, it is a homo-dimer when most of photo-inducible dimers are heterodimers. In addition, the use of VVD is easy; and doesn???t need any addition of co-factors: VVD works with Flavin adenine dinucleotide (FAD) which is already abundant in eukaryote and prokaryote cells (M??ller & Weber, 2013; Nihongaki, Suzuki, Kawano, & Sato, 2014). false false _2033_ 22805 22805 9 false We found illegal sites (PstI) into VVD sequence, those one have been removed. false Johanna Chesnel annotation2470046 1 VVD range2470046 1 1 450 BBa_J61100 1 BBa_J61100 Ribosome Binding Site Family Member 2007-01-28T12:00:00Z 2015-08-31T02:03:00Z N/A {{JCA_Arkin_RBSFamily}} false true _95_ 0 483 95 In stock false N/A true John Anderson BBa_K1616016 1 YFP-C YC155 - C terminal YFP split 2015-09-17T11:00:00Z 2016-01-21T02:25:51Z This part works with BBa_K16160017. A split protein is a protein whose sequence has been divided into two (or more) different parts. Often used to study protein-protein interactions, the protein can not perform its function until the parts are put back together. For instance, YFP, the yellow-fluorescent protein, will only express fluorescence when its two parts will be reunited. In normal condition, the production of a protein in response to a stimulus can easily reach several hours due to the many steps required for the protein synthesis. By using split-proteins, we are taking advantage of the absence of fluorescence when the two parts are apart. Indeed, the two parts of our split-YFP, when remaining separated, can be produced without being effective. Therefore, the overall process is far less time-consuming. However, to implement a light control on the fluorescence activation, a genetic construction gathering the VVD photoreceptor and our split-YFP has to be engineered. The new alternative approach for the visualization of protein interactiosn has been developed; the biomolecular fluorescence complementation (BiFC) techniques based on the complementation between fragments of fluorescent proteins; fragments of the yellow fluorescent protein (YFP) brought together by the association of two interaction partners fused to the fragments. They noticed that the spectral characteristics of BiFC of YFP were virtually identical to those of intact YFP.(Chang-Deng Hu, 2003) false false _2033_ 4206 22805 9 false Illegal sites have been checked. false Johanna Chesnel annotation2470135 1 YC155 range2470135 1 1 255 BBa_I712074 1 BBa_I712074 T7 promoter (strong promoter from T7 bacteriophage) 2007-10-21T11:00:00Z 2015-08-31T04:07:46Z T7 bacteriophage T7 promoter is very specific promoter which is transcribed only by specific T7 RNA polymerase. Usually this promoter is used in expression systems where T7 promoter is cotransfected with T7 RNA polymerase. That ensures strong transcription of desired genes. false false _130_ 0 1856 9 In stock false true Rok Gaber BBa_K1616001 1 VVD-YC VVD linked to YC155 (YFP Cter split) with promoter T7 2015-09-16T11:00:00Z 2015-09-22T06:42:02Z This part have been created thank to gblock, our team have assembled the sequence of photoreceptor VVD (without illegal site), a linker(1) and then the C terminal of YFP split(1). (1) Tom Kerppola, Ph. D, investigator at the Howard Hughes Medical Institute as well as Professor in the University of Michigan Vivid (VVD) is the smallest known Light???oxygen???voltage (LOV) domain protein and photo-inducible dimer. Isolated from Neurospora crassa, VVD forms a homo-dimer in response to a blue-light stimulus. Then, a split protein is a protein whose sequence has been divided into two (or more) different parts. The yellow-fluorescent (YFP) protein will only express fluorescence when its two parts will be reunited. The Part is coding for the homodimer VVD link by an integration of specific sequence to the C terminal of the YFP split. So, this part works with BBa_K1616002. In absence of blue-light, the conformation of the VVD photoreceptor will prevent the formation of the complete fluorescent protein while in presence of the light signal the YFP protein will be reconstituted leading to the fast expression of a yellow fluorescence in our bacteria. false false _2033_ 22805 22805 9 true The sequence of VVD had 2 illegal sites PstI; that have been removed. false Johanna Chesnel component2471224 1 BBa_K1616021 component2471217 1 BBa_J61100 component2471216 1 BBa_I712074 annotation2471224 1 BBa_K1616021 range2471224 1 75 830 annotation2471217 1 BBa_J61100 range2471217 1 55 66 annotation2471216 1 BBa_I712074 range2471216 1 1 46 BBa_K1616015 1 Linker-FOS Linker FOS of 17 aa 2015-09-17T11:00:00Z 2015-09-18T03:09:19Z Linker used in association with VVD (BBa_K1616014) This part is a sequence coding for 17 aa useful to link two proteins false false _2033_ 22805 22805 9 false Illegal sites have been checked false Johanna Chesnel annotation2470111 1 linker range2470111 1 1 51 BBa_K1616021 1 VVD-YC VVD linked to YC155 (YFP Cter split) 2015-09-17T11:00:00Z 2015-09-18T05:28:45Z Assembly of 3 parts: BBa_K1616014, BBa_K1616015 and BBa_K1616016 without scar See BBa_K1616001. This part have been created in order to have the three parts without scars (VVD,linker and YFP Nterminal). false false _2033_ 22805 22805 9 false No separation between the 3 parts. false Johanna Chesnel component2471213 1 BBa_K1616015 component2471215 1 BBa_K1616016 component2471211 1 BBa_K1616014 annotation2471211 1 BBa_K1616014 range2471211 1 1 450 annotation2471215 1 BBa_K1616016 range2471215 1 502 756 annotation2471213 1 BBa_K1616015 range2471213 1 451 501 BBa_K1616021_sequence 1 cacaccctgtatgcccctggcggctacgacatcatgggctacctgatccagatcatgaagcggcccaacccccaggtggaactgggccctgtggatacctctgtggccctgatcctgtgcgacctgaagcagaaagacacccccatcgtgtacgcctccgaggccttcctgtacatgaccggctactccaacgccgaggtgctgggccggaactgtagattcctccagtcccctgacggcatggtcaagcctaagtccacccggaaatacgtggactctaacaccatcaacaccatgcggaaggccatcgaccggaacgctgaggtgcaggtggaagtcgtgaacttcaagaagaacggccagcgcttcgtgaatttcctgaccatgatccccgtgcgggacgagacaggcgagtacagatactccatgggcttccagtgcgagacagagcgtccggcgtgcaaaatcccgaacgacctgaaacagaaagtcatgaaccacgacaagcagaagaacggcatcaaggtgaacttcaagatccgccacaacatcgaggacggcagcgtgcagctcgccgaccactaccagcagaacacccccatcggcgacggccccgtgctgctgcccgacaaccactacctgagctaccagtccgccctgagcaaagaccccaacgagaagcgcgatcacatggtcctgctggagttcgtgaccgccgccgggatcactctcggcatggacgagctgtacaagtaa BBa_K1616001_sequence 1 taatacgactcactatagggaatacaagctacttgttctttttgcatactagagaaagaggggacatactagagcacaccctgtatgcccctggcggctacgacatcatgggctacctgatccagatcatgaagcggcccaacccccaggtggaactgggccctgtggatacctctgtggccctgatcctgtgcgacctgaagcagaaagacacccccatcgtgtacgcctccgaggccttcctgtacatgaccggctactccaacgccgaggtgctgggccggaactgtagattcctccagtcccctgacggcatggtcaagcctaagtccacccggaaatacgtggactctaacaccatcaacaccatgcggaaggccatcgaccggaacgctgaggtgcaggtggaagtcgtgaacttcaagaagaacggccagcgcttcgtgaatttcctgaccatgatccccgtgcgggacgagacaggcgagtacagatactccatgggcttccagtgcgagacagagcgtccggcgtgcaaaatcccgaacgacctgaaacagaaagtcatgaaccacgacaagcagaagaacggcatcaaggtgaacttcaagatccgccacaacatcgaggacggcagcgtgcagctcgccgaccactaccagcagaacacccccatcggcgacggccccgtgctgctgcccgacaaccactacctgagctaccagtccgccctgagcaaagaccccaacgagaagcgcgatcacatggtcctgctggagttcgtgaccgccgccgggatcactctcggcatggacgagctgtacaagtaa BBa_K1616014_sequence 1 cacaccctgtatgcccctggcggctacgacatcatgggctacctgatccagatcatgaagcggcccaacccccaggtggaactgggccctgtggatacctctgtggccctgatcctgtgcgacctgaagcagaaagacacccccatcgtgtacgcctccgaggccttcctgtacatgaccggctactccaacgccgaggtgctgggccggaactgtagattcctccagtcccctgacggcatggtcaagcctaagtccacccggaaatacgtggactctaacaccatcaacaccatgcggaaggccatcgaccggaacgctgaggtgcaggtggaagtcgtgaacttcaagaagaacggccagcgcttcgtgaatttcctgaccatgatccccgtgcgggacgagacaggcgagtacagatactccatgggcttccagtgcgagacagag BBa_K1616015_sequence 1 cgtccggcgtgcaaaatcccgaacgacctgaaacagaaagtcatgaaccac BBa_J61100_sequence 1 aaagaggggaca BBa_I712074_sequence 1 taatacgactcactatagggaatacaagctacttgttctttttgca BBa_K1616016_sequence 1 gacaagcagaagaacggcatcaaggtgaacttcaagatccgccacaacatcgaggacggcagcgtgcagctcgccgaccactaccagcagaacacccccatcggcgacggccccgtgctgctgcccgacaaccactacctgagctaccagtccgccctgagcaaagaccccaacgagaagcgcgatcacatggtcctgctggagttcgtgaccgccgccgggatcactctcggcatggacgagctgtacaagtaa igem2sbol 1 iGEM to SBOL conversion Conversion of the iGEM parts registry to SBOL2.1 Chris J. Myers James Alastair McLaughlin 2017-03-06T15:00:00.000Z