BBa_K1933200 1 BBa_K1933200 constitutive expression of anti-Norovirus GII.4 scFv fused to INPNC with 6xHis tag 2016-10-09T11:00:00Z 2016-10-17T04:57:35Z Promoter and RBS The INPNC surface expression component derives from Pseudomonas Syringae. We used parts BBa_K811005 from iGEM12_Penn. scFv is made from fusing variable region (VH and VL chain) of a human anti-norovirus antibody with a linker peptide. We have been given the scFv-encoding plasmid (NoV GII.4 strain-specific antibody 12A2 (vector: pSCCA5-E8d)) from Dr. Kyoko Moriguchi of the Fujita Health University Department of Virology Parasitology. anti-Norovirus GII.4 scFv fused to INPNC with 6xHis tag is one of a series of surface expressing fusion proteins that make up biodevice that aims to be therapeutic solution against norovirus infections. This protein in particular is a single chained variable fragment(scFv) of a norovirus GII.4 strain fused to surface expression anchoring domain(INPNC), connected by a poly-histidine tag to be easily identified by Western blotting. For more information, please visit our <a ref=http://2016.igem.org/Team:Kyoto> Wiki page</a>. false false _2400_ 23701 29998 9 true Silent mutation was done to remove PstI site. false Tomoki Uchino component2515638 1 BBa_K1933002 component2515627 1 BBa_K1933004 component2515631 1 BBa_K1933011 annotation2515638 1 BBa_K1933002 range2515638 1 1036 1815 annotation2515627 1 BBa_K1933004 range2515627 1 1 60 annotation2515631 1 BBa_K1933011 range2515631 1 61 1035 BBa_K1933011 1 BBa_K1933011 INPNC fused to 6xHis tag without stop codon 2016-10-16T11:00:00Z 2016-10-17T01:49:52Z INPNC derived from BBa_K811005 INPNC codes for N- and C- terminal domain of Ice-Nucleation Protein (INP) from Pseudomonas syringae , connected by a 6xHis tag to be easily identified by Western blotting[1]. This is used as an anchoring motif for the cell surface display of passengers. This domain does not have signal peptide for anchoring on the cell surface[2]. Surface displayed passengers retain enzyme activity even when it is linked to INPNC with 6xHis tag. false false _2400_ 23701 23701 9 false We omitted stop codon from BBa_K1933001 to use it for the registration of composite parts. false Sora Matsumoto, Tomoki Uchino annotation2514865 1 start codon range2514865 1 1 3 annotation2514868 1 6xHis tag range2514868 1 958 975 annotation2514867 1 INPNC range2514867 1 4 957 BBa_K1933002 1 scFv anti-Norovirus GII.4 scFv 2016-10-09T11:00:00Z 2016-10-18T10:28:59Z We have been given given the scFv-encoding plasmid (NoV GII.4 strain-specific antibody 12A2 (vector: pSCCA5-E8d)) from Dr. Kyoko Moriguchi of the Fujita Health University Department of Virology Parasitology. This part codes anti-Norovirus GII.4 scFv (single-chain variable fragment) which derives from human 12A2 antibody. This scFv is suggested to prevent binding of human norovirus to Hist-Blood Group Antigens. [1] This coding sequence was fused to INPNC-His (part number: ) or BclA-His (part number: ) in our project. For more information, please visit our <a ref=http://2016.igem.org/Team:Kyoto> Wiki page</a>. false false _2400_ 23701 29998 9 true silent mutation was made to remove PstI: G255C false Tomoki Uchino annotation2490182 1 start codon range2490182 1 1 3 annotation2490183 1 VH range2490183 1 4 392 annotation2490185 1 VL range2490185 1 439 777 annotation2490184 1 linker range2490184 1 393 438 annotation2490186 1 stop codon range2490186 1 778 780 annotation2490197 1 G255C range2490197 1 255 255 BBa_K1933004 1 BBa_K1933004 constitutive promoter and RBS 2016-10-09T11:00:00Z 2016-10-10T09:38:21Z BBa_J23105 was designed by John Anderson and BBa_B0034 was designed by Vinay S Mahajan, Voichita D. Marinescu, Brian Chow, Alexander D Wissner-Gross and Peter Carr IAP, 2003. This sequence was chemically synthesized. This part codes costitutive promoter(BBa_J23105) and RBS(BBa_B0034). Most parts were expressioned by this part in our project. false false _2400_ 29998 29998 9 false 7bp after promoter and 6bp after RBS false Tomoki Uchino annotation2490322 1 nonsense sequence range2490322 1 36 42 annotation2490323 1 Re-entry of BBa_J23105 range2490323 1 1 35 annotation2490408 1 constitutive promoter and RBS range2490408 1 1 60 annotation2490405 1 scar range2490405 1 55 60 annotation2490404 1 Re-entry of BBa_B0034 range2490404 1 43 54 BBa_K1933200_sequence 1 tttacggctagctcagtcctaggtactatgctagcactagagaaagaggagaaatactagatgaccctggacaaagctctggttctgcgtacctgcgctaacaacatggctgaccactgcggtctgatctggccggcttctggtaccgttgaatctcgttactggcagtctacccgtcgtcacgaaaacggtctggttggtctgctgtggggtgctggtacctctgctttcctgtctgttcacgctgacgctcgttggatcgtttgcgaagttgctgttgctgacatcatctctctggaagaaccgggtatggttaaattcccgcgtgctgaagttgttcacgttggtgaccgtatctctgcttctcacttcatctctgctcgtcaggctgacccggcttctacctctacctctacctctacctctaccctgaccccgatgccgaccgctatcccgaccccgatgccggctgttgcttctgttaccctgccggttgctgaacaggctcgtcacgaagttttcgacgttgcttctgtttctgctgctgctgctccggttaacaccctgccggttaccaccccgcagaacctgcaaaccgctacctacggttctaccctgtctggtgacaaccactctcgtctgatcgctggttacggttctaacgaaaccgctggtaaccactctgacctgatcggtggtcacgactgcaccctgatggctggtgaccagtctcgtctgaccgctggtaaaaactctgttctgaccgctggtgctcgttctaaactgatcggttctgaaggttctaccctgtctgctggtgaagactctaccctgatcttccgtctgtgggacggtaaacgttaccgtcagctggttgctcgtaccggtgaaaacggtgttgaagctgacatcccgtactacgttaacgaagacgacgacatcgttgacaaaccggacgaagacgacgactggatcgaagttaaaggtggcggtagtggcggtgcaggaggcggatcccatcatcatcatcatcatatggccgaggtgcagctggtggagtctgggggaggcttggtacagcctggggggtccctgagactctcctgtgcaacctctgcattcatctttgacaactatcccatgacttgggtccgccaggctcaggctccagggaaggggctggagtgggtcgcaagtattagtggtagtggcggcaggacatactacgcagactccgtgaagggccgcttcaccatctccagagacaattccaagtatgtgttgtttctccagatgaacagcctgagagccgaggacacggccatatattactgtgtgaaatctcctcccgacgtttgggggagtcatcaattcagggattggtactttgactactggggccagggaaccctggtcaccgtctcgagaggcggtggcggatcaggtggcggtggaagtggcggtggtgggtccatggcctcctatgtgctgactcagccaccctcagcgtctgggacccccgggcagagggtcaccatctcttgttctggaagcagctccaacatcggaagtaatactataaactggtaccagcagctcccaggaacggcccccaaactcctcatctataataatcatcagcggccctcaggggtccctgaccgattctctggctcaaagtctggcacctcagcctccctggccatcagtgggctccagtctgcggatgaggctgattattactgtggagcgtggaatgacagcctgaatgtctatgtcttcggaactgggaccaaggtcaccgtcctaggttag BBa_K1933002_sequence 1 atggccgaggtgcagctggtggagtctgggggaggcttggtacagcctggggggtccctgagactctcctgtgcaacctctgcattcatctttgacaactatcccatgacttgggtccgccaggctcaggctccagggaaggggctggagtgggtcgcaagtattagtggtagtggcggcaggacatactacgcagactccgtgaagggccgcttcaccatctccagagacaattccaagtatgtgttgtttctccagatgaacagcctgagagccgaggacacggccatatattactgtgtgaaatctcctcccgacgtttgggggagtcatcaattcagggattggtactttgactactggggccagggaaccctggtcaccgtctcgagaggcggtggcggatcaggtggcggtggaagtggcggtggtgggtccatggcctcctatgtgctgactcagccaccctcagcgtctgggacccccgggcagagggtcaccatctcttgttctggaagcagctccaacatcggaagtaatactataaactggtaccagcagctcccaggaacggcccccaaactcctcatctataataatcatcagcggccctcaggggtccctgaccgattctctggctcaaagtctggcacctcagcctccctggccatcagtgggctccagtctgcggatgaggctgattattactgtggagcgtggaatgacagcctgaatgtctatgtcttcggaactgggaccaaggtcaccgtcctaggttag BBa_K1933004_sequence 1 tttacggctagctcagtcctaggtactatgctagcactagagaaagaggagaaatactag BBa_K1933011_sequence 1 atgaccctggacaaagctctggttctgcgtacctgcgctaacaacatggctgaccactgcggtctgatctggccggcttctggtaccgttgaatctcgttactggcagtctacccgtcgtcacgaaaacggtctggttggtctgctgtggggtgctggtacctctgctttcctgtctgttcacgctgacgctcgttggatcgtttgcgaagttgctgttgctgacatcatctctctggaagaaccgggtatggttaaattcccgcgtgctgaagttgttcacgttggtgaccgtatctctgcttctcacttcatctctgctcgtcaggctgacccggcttctacctctacctctacctctacctctaccctgaccccgatgccgaccgctatcccgaccccgatgccggctgttgcttctgttaccctgccggttgctgaacaggctcgtcacgaagttttcgacgttgcttctgtttctgctgctgctgctccggttaacaccctgccggttaccaccccgcagaacctgcaaaccgctacctacggttctaccctgtctggtgacaaccactctcgtctgatcgctggttacggttctaacgaaaccgctggtaaccactctgacctgatcggtggtcacgactgcaccctgatggctggtgaccagtctcgtctgaccgctggtaaaaactctgttctgaccgctggtgctcgttctaaactgatcggttctgaaggttctaccctgtctgctggtgaagactctaccctgatcttccgtctgtgggacggtaaacgttaccgtcagctggttgctcgtaccggtgaaaacggtgttgaagctgacatcccgtactacgttaacgaagacgacgacatcgttgacaaaccggacgaagacgacgactggatcgaagttaaaggtggcggtagtggcggtgcaggaggcggatcccatcatcatcatcatcat 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