BBa_K597101 1 BBa_K597101 GFP-AviTag [AviTagged green fluorescent protein] 2011-09-25T11:00:00Z 2015-05-08T01:12:50Z GFP aliquot was provided by Dr. DeLisa (Cornell University, Chemical Engineering Department) in the form of a pZE12-SMCS vector. AviTag: Avidity, L.L.C., Eleanor Roosevelt Institute, Denver, Colorado 80206, USA. Ref: Cull MG, Schatz PJ (2000). ???Biotinylation of proteins in vivo and in vitro using small peptide tags.??? | Methods of Enzymology Vol 326, 2000, Pages 430-440. GFP-avitag codes for green fluorescent protein with an appended biotinylation marker. When attached to a promoter, it can be used as a fluorescent reporter and biotinylated by E coli. E coli has a natural mechanism encoded by the BirA gene to produce biotin and biotinylate proteins - this mechanism can be enhanced by culturing with biotin. This enzyme can then be used in biotin-streptavidin binding applications. The GFP-avitag fusion protein was constructed to have the AviTag sequence on the C-terminus, followed by a stop codon. The resulting protein, once biotinylated, is capable of a ligand reaction with surface-coated avidin, while retaining the fluorescence of GFP. This molecule was used to test the efficacy of the binding interaction between the functionalized protein and a streptavidin-coated surface. false false _769_ 0 10191 9 It's complicated true The avitag is capable of being on either the C or N terminus of the protein of interest with little difference between the two in the binding efficiency or expression of the protein. We chose to put the Avitag on the C-terminus for the convenience of using a primer dimer pair to insert the iGEM suffix to the sample provided by the Delisa Lab Group. false James Mathew annotation2142926 1 Start range2142926 1 1 3 annotation2142928 1 AviTag range2142928 1 721 765 annotation2142927 1 GFP range2142927 1 1 714 annotation2142930 1 AgeI range2142930 1 715 720 annotation2142929 1 Stop range2142929 1 766 768 BBa_K597101_sequence 1 atgagtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcactactttcgcgtatggtcttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcatggcagacaaacaaaagaatggaatcaaagttaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggattacacatggcatggatgaactatacaaaaccggtggcctgaacgacatcttcgaggctcagaaaatcgaatggcacgaataa 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