BBa_K1989028 1 BBa_K1989028 Triple spytag with mSA, His-tag , FlAsH-tag and signal peptide PhoA 2016-10-11T11:00:00Z 2016-10-12T09:09:13Z Inspiring from the self-catalysis of isopeptide bond between Lys and Asp in Streptococcus pyogenes fibronectin-binding protein FbaB, researchers split the catalytic domain and obtain two peptide called Spytag(the short one) and Spycatcher(the long one) which are able to form isopeptide bond with the other without any assistant. By fusing Spytag and Spycatcher with functional domains respectively, researchers solve the problem tactfully. In order to using Spytag and Spycatcher system as scaffold, we fused three Spytag spaced by (VPGVG)4 with 6xHistag in N-terminal and another functional protein called monomeric streptavidin(mSA) in C-terminal. The fused protein PhoA-Histag-3A-mSA-FlAsHtag possess both isopeptide bond forming function, secretion and collectable. Thus, using PhoA-Histag-3A- mSA -FlAsHtag as a part of hydrogel formation, we can obtain our multifunctional biomaterial. false false _2456_ 17187 17187 9 false The length of the linker between spytag may affect the physical and chemical property of hydrogel. As several attributes of our target proteins, especially their folding state in cells, we were unable to rationally design or choose a signal peptide for each. Nevertheless, we could build a Signal Peptide Repertory to screen for the best SP candidate. Our ultimate goal was to select the most appropriate ones from a huge amount of signal peptides. false Yiming Dong annotation2493907 1 mSA range2493907 1 466 824 annotation2493906 1 Linker range2493906 1 382 461 annotation2493905 1 Spytag-Spytag-Spytag range2493905 1 130 377 annotation2493910 1 FlAsH-tag range2493910 1 832 870 annotation2493903 1 Signal peptide PhoA range2493903 1 24 90 annotation2493904 1 His-tag range2493904 1 91 129 BBa_K1989028_sequence 1 tggtttaactttaagaaggagatatacatgaaacagtctaccatcgctctggctctgctgccgctgctgttcaccccggttaccaaagctatgaaaggttcttctcatcatcatcatcatcacgttgacgcccatattgttatggtggatgcatataaaccgaccaaactggatggtcatggtgtgggtgttccgggtgttggtgtgcctggcgttggcgttcctggtgtgggcgaactggcacatatcgtgatggttgatgcctacaaacctacaaaaaccagcgtgccaggtgttggagtaccgggtgtaggcgttccaggtgaaggtggtctgctggatgcgcacattgtaatggttgacgcgtataaaccaacgaaaaccagcgttccgggtgttggtgttcctggtgaaggtgtgcctggtgtgggtgtaccaggcgtaggtgttccaggtggtgaactggcggaggcgggtatcaccggcacctggtacaaccagcacggtagcaccttcaccgtgaccgcgggtgcggacggtaacctgaccggtcaatatgaaaaccgtgcgcagggtaccggctgccaaaacagcccgtacaccctgaccggtcgttataacggcaccaagctggagtggcgtgttgaatggaacaacagcaccgagaactgccacagccgtaccgaatggcgtggccagtaccaaggtggcgcggaggcgcgtattaacacccagtggaacctgacctatgagggtggcagcggtccggcgaccgaacagggccaagatacctttaccaaggtgaaaccgagcgcggcgagcggcagttccttcctgaactgctgcccgggttgctgcatggaaccgtaacaaagcccgaaaggaagctgagttggcttaa 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