BBa_K1947022 1 BBa_K1947022 Improve the protein purification. 2016-10-13T11:00:00Z 2016-10-17T10:55:52Z Protein splicing elements (termed inteins) were described as in-frame insertions in the Saccharomyces cerevisiae VMA1 gene. TEV protease may bring some impure proteins. We use intein to replace the TEV site. The M. xenopi GyrA intein provides a paradigm for a minimal protein splicing element. Intein can be cleaved by DTT. DTT as reducing agent protects the free thiol groups of the protein from oxidation and so it doesn???t bring foreign substance. As a result, intein can achieve a better effect of purification than TEV protease. true false _2414_ 33797 33797 9 false GS linker is used to construct the fusion protein. Amilcp does not have a termination  codon. false Jie Tang component2500791 1 BBa_K592009 component2500792 1 BBa_K1947002 component2500793 1 BBa_K1947020 annotation2500793 1 BBa_K1947020 range2500793 1 1287 1325 annotation2500791 1 BBa_K592009 range2500791 1 1 669 annotation2500792 1 BBa_K1947002 range2500792 1 676 1278 BBa_K592009 1 amilCP amilCP, blue chromoprotein 2011-09-17T11:00:00Z 2015-05-08T01:12:48Z Acropora millepora Released HQ 2013 This chromoprotein, amilCP, naturally exhibits very strong color when expressed. The color is blue/purple and is visible to naked eye, thereby requiring no instruments to observe. This DNA was provided by Jeffrey Miller at UCLA. It was made BioBrick-compatible after removal of one illegal internal restriction site (EcoRI). false false _763_ 0 7929 9 In stock true Illegal internal restriction site had to be removed (EcoRI). false Lei Sun annotation2131628 1 amilCP range2131628 1 1 666 BBa_K1947002 1 Intein Intein 2016-10-12T11:00:00Z 2016-10-14T02:59:19Z This sequence comes form the gyrA gene of Mycobacterium xenopi This sequence encodes a short polypeptide which can be specifically recognized and spliced by DL-Dithiothreitol（DTT）. In our case we add this short sequence between the recombinant protein which we want to purify and the linker protein Spytag, in order to separate the recombinant protein form the Magnetosome. false false _2414_ 33528 34115 9 false Our project is to use Magnetosome to purify recombinant protein, and we need to remove the protein from the compound after the protein we are interested combined with Magnetosome. As this short polypeptide can be spliced by DTT, we can meet this need by adding the intein between the recombinant protein and magnetosome. false zhang xucheng BBa_K1947020 1 BBa_K1947020 This polypeptide will be fused with the recombinant protein which we want to purify. 2016-10-13T11:00:00Z 2016-10-14T01:54:38Z Streptococcus pyogenes fibro-nectin-binding protein FbaB contains a domain with a spontaneous isopeptide bond between Lys31 and Asp117. This domain can be splitted into two fragments: the small one is named as Spytag, while the big one is Spycatcher. Spytag is separated from the second immunoglobulin-like collagen adhesin domain (CnaB2) which is come from the fibronectin binding protein FbaB. There is a highly specific recognition and covalent conjugation between Spytag (a peptide with 13 amino acids) and Spycatcher (a small protein consisting of 138 residues). A protein of interest with a Spytag at its N- or C-terminus is expressed in E. coli and present in bacterial lysate.This part encodes a polypeptide that can combined with Spycatcher, through a covalent bond. In our case this polypeptide will be fused with the recombinant protein which we want to purify. true false _2414_ 33797 33797 9 false This part will be fused with the recombinant protein which we want to purify. false Jie Tang BBa_K1947022_sequence 1 atgagtgtgatcgctaaacaaatgacctacaaggtttatatgtcaggcacggtcaatggacactactttgaggtcgaaggcgatggaaaaggtaagccctacgagggggagcagacggtaaagctcactgtcaccaagggcggacctctgccatttgcttgggatattttatcaccacagtgtcagtacggaagcataccattcaccaagtaccctgaagacatccctgactatgtaaagcagtcattcccggagggctatacatgggagaggatcatgaactttgaagatggtgcagtgtgtactgtcagcaatgattccagcatccaaggcaactgtttcatctaccatgtcaagttctctggtttgaactttcctcccaatggacctgtcatgcagaagaagacacagggctgggaacccaacactgagcgtctctttgcacgagatggaatgctgctaggaaacaactttatggctctgaagttagaaggaggcggtcactatttgtgtgaatttaaaactacttacaaggcaaagaagcctgtgaagatgccagggtatcactatgttgaccgcaaactggatgtaaccaatcacaacaaggattacacttcggttgagcagtgtgaaatttccattgcacgcaaacctgtggtcgcctaataatactagatgtgcatcacgggagatgcgctggttgccctacccgagggcgagtcggtacgcatcgccgacatcatcccgtgctcgccgaccggctgttccactccggcgagcatccggtgtacacggtgcgtacggtcggtgccgggtgcgcggcccaacagtgacaacgccatcgacctgaaagtccttgaccggcatggcaaaggtctgcgtgtgacgggcaccgcgaaccacccgttgttgtgtttggtcgacgtcgccggggtgccgaccctgctgtggaagctgatcgacgaaatcaagccgggcgattacgcggtgattcaacgcagcgcattcagcgtcgactgtgcaggttttgcccgcggaaaacccgaatttgcgcccacaacctacacagtcggcgtccctggactggtgcgtttcttggaagcacaccaccgagacccggacgcccaagctatcgccgacgagctgaccgacgggcggttctactacgcgaaagtcgccagtgtcaccgacgccggcgtgcagccggtgtatagccttcgtgtcgacacggcagaccacgcgtttatcaccaacgggttcgtcagccacgcgtaataatactagaggctcatattgtcatggttgatgcttacaagccaactaag BBa_K1947020_sequence 1 gctcatattgtcatggttgatgcttacaagccaactaag BBa_K592009_sequence 1 atgagtgtgatcgctaaacaaatgacctacaaggtttatatgtcaggcacggtcaatggacactactttgaggtcgaaggcgatggaaaaggtaagccctacgagggggagcagacggtaaagctcactgtcaccaagggcggacctctgccatttgcttgggatattttatcaccacagtgtcagtacggaagcataccattcaccaagtaccctgaagacatccctgactatgtaaagcagtcattcccggagggctatacatgggagaggatcatgaactttgaagatggtgcagtgtgtactgtcagcaatgattccagcatccaaggcaactgtttcatctaccatgtcaagttctctggtttgaactttcctcccaatggacctgtcatgcagaagaagacacagggctgggaacccaacactgagcgtctctttgcacgagatggaatgctgctaggaaacaactttatggctctgaagttagaaggaggcggtcactatttgtgtgaatttaaaactacttacaaggcaaagaagcctgtgaagatgccagggtatcactatgttgaccgcaaactggatgtaaccaatcacaacaaggattacacttcggttgagcagtgtgaaatttccattgcacgcaaacctgtggtcgcctaataa BBa_K1947002_sequence 1 atgtgcatcacgggagatgcgctggttgccctacccgagggcgagtcggtacgcatcgccgacatcatcccgtgctcgccgaccggctgttccactccggcgagcatccggtgtacacggtgcgtacggtcggtgccgggtgcgcggcccaacagtgacaacgccatcgacctgaaagtccttgaccggcatggcaaaggtctgcgtgtgacgggcaccgcgaaccacccgttgttgtgtttggtcgacgtcgccggggtgccgaccctgctgtggaagctgatcgacgaaatcaagccgggcgattacgcggtgattcaacgcagcgcattcagcgtcgactgtgcaggttttgcccgcggaaaacccgaatttgcgcccacaacctacacagtcggcgtccctggactggtgcgtttcttggaagcacaccaccgagacccggacgcccaagctatcgccgacgagctgaccgacgggcggttctactacgcgaaagtcgccagtgtcaccgacgccggcgtgcagccggtgtatagccttcgtgtcgacacggcagaccacgcgtttatcaccaacgggttcgtcagccacgcgtaataa 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