BBa_K416003 1 BBa_K416003 Yeast Secretion Tag 2010-06-22T11:00:00Z 2015-05-08T01:12:27Z This part was sent to use from Dr. Sheldon Park of the University of Buffalo. This secretion tag when linked to the N-terminal of the protein directs extracellular secretion of the protein. After targeting through the ER to the golgi, this "pre-pro" tag is cleaved by the KEX-2 protease resulting in secretion of protein without the aforementioned tag. To use this tag, attached it directly upstream of your proteins CDS. false false _527_ 0 7430 9 It's complicated true There was an illegal Xbal1 site at bp 101 and we removed it through altering the primers necessary for biobricking the sequence. false John Phair BBa_K1772004 1 BBa_K1772004 yEGFP-tagged Lignin peroxidase 2015-09-17T11:00:00Z 2015-09-17T11:41:49Z Promoter: BBa_I766555 Kozak sequence: BBa_J63003 yEGFP (yeast Enhanced Green Fluorescent Protein): NCBI GenBank accession U73901.1 Linker sequences: BBa_K243004 Lignin Peroxidase from Reticulitermes flavipes, NCBI GenBank accession JX513905.1 Secretion tag: Bba_K416003 Terminator: BBa_K392003 Lignin Peroxidase fusion protein gene flanked by two linker sequences, with a a GFP tag on the N-terminus and a yeast secretion tag on the C-terminus. A constitutive medium strength pCyc promoter followed by a yeast kozak sequence caps the N-terminus, and a yeast terminator sequence caps the C-terminus. false false _2194_ 22695 22695 9 false Our goal was to synthesize lignin-degrading enzymes to be secreted out of yeast cells, which we used the secretion tag for. The fluorescence tag was merely implemented to simplify detection of successful secretion. To avoid time-consuming and low-efficiency standard assembly techniques, this construct was designed to be synthesized by IDT using two GBlocks(c), which were to be joined by Gibson Assembly. Linker sequences were implemented in order to minimize interaction between protein domains. A constitutive promoter was implemented due to the intended application (a bioreactor pretreatment process where a yeast containing this part would secrete enzymes to continuously degrade lignin). false Tianyu Tan component2469434 1 BBa_K392003 component2469427 1 BBa_J63003 component2469430 1 BBa_K1772003 component2469433 1 BBa_K416003 component2469429 1 BBa_K243004 component2469425 1 BBa_I766555 component2469432 1 BBa_K243004 annotation2469433 1 BBa_K416003 range2469433 1 1847 1960 annotation2469427 1 BBa_J63003 range2469427 1 253 270 annotation2469429 1 BBa_K243004 range2469429 1 279 290 annotation2469432 1 BBa_K243004 range2469432 1 1829 1840 annotation2469425 1 BBa_I766555 range2469425 1 1 244 annotation2469434 1 BBa_K392003 range2469434 1 1969 2097 annotation2469430 1 BBa_K1772003 range2469430 1 297 1820 BBa_K243004 1 ShortLinke Short Linker (Gly-Gly-Ser-Gly) 2009-10-11T11:00:00Z 2015-05-08T01:11:37Z Oligos synthesized by sigma. Hybridized by PCR. This linker was used to connect two different parts. The sequence produced the aminoacids Gly-Gly-Ser-Gly. false true _352_ 0 4732 9 In stock false None. false Freiburg Bioware09 annotation2041881 1 Short Linker range2041881 1 1 12 BBa_I766555 1 BBa_I766555 pCyc (Medium) Promoter 2007-08-15T11:00:00Z 2015-08-31T04:08:10Z Endogenous promoter for Cytochrome c Mid-expression level constitutive promoter in yeast false false _155_ 0 1931 9 Not in stock false none false Nili Sommovilla BBa_K1772003 1 BBa_K1772003 Lignin Peroxidase 2015-09-17T11:00:00Z 2015-09-17T11:41:36Z Reticulitermes flavipes (GenBank Accession JX513905.1) Protein coding sequence of Lignin Peroxidase, an enzyme that is the initiator and one of the main enzymes responsible for the breakdown of lignin inside the cell walls of plant cells. Reticulitermes flavipes, or the eastern subterranean termite, uses this enzyme to digest woody debris with higher efficiency. Lignin peroxidase has the ability to oxidize aromatic compounds in lignin with redox potentials above 1.4 eV, which is the equivalent to reducing metals like gold or molecules such as lead oxide. It is believed that this action is done by catalyzing the transfer of an oxygen atom from a hydrogen peroxide molecule to an aromatic ring. However, the exact redox mechanism for lignin peroxidase is currently unknown. false false _2194_ 22695 22695 9 false This gene sequence excludes introns (noncoding segments) present in the original gene from Reticulitermes flavipes. Sequence optimization was done using IDT's codon optimizer. Illegal restriction enzyme sites (under RFC 10 definition) were detected using NEBcutter2 and removed with silent mutations. false Tianyu Tan BBa_K392003 1 BBa_K392003 yeast ADH1 terminator 2010-10-09T11:00:00Z 2015-05-08T01:12:20Z Registry parts BBa_K105027 (promoter) and BBa_J63003 (Kozak). Engineered 'cyc100 promoter' of <i>Saccharomyces cerevisiae</i> with Kozak sequence attached downstream. false false _528_ 0 6220 9 It's complicated true Assembled by 3A method. false Tadashi Nakamura, Shuhei Yasumoto, Takahiro Saka, Saya Kakuda BBa_J63003 1 Kozak & st designed yeast Kozak sequence 2006-10-10T11:00:00Z 2015-08-31T01:56:26Z consensus Kozak and start built from oligonucleotides Released HQ 2013 consensus Kozak sequence and start codon false false _97_ 0 545 97 In stock false designed such that fusion using fusion bricks assembly method leads to in frame translation true Caroline Ajo-Franklin annotation1902852 1 start codon range1902852 1 13 15 BBa_K243004_sequence 1 ggtggttctggt BBa_I766555_sequence 1 cagatccgccaggcgtgtatatatagcgtggatggccaggcaactttagtgctgacacatacaggcatatatatatgtgtgcgacgacacatgatcatatggcatgcatgtgctctgtatgtatataaaactcttgttttcttcttttctctaaatattctttccttatacattaggacctttgcagcataaattactatacttctatagacacacaaacacaaatacacacactaaattaata BBa_K416003_sequence 1 atgaaagttttgattgttttgttggctattttcgctgctttgccattggctttggctcaaccagttatttctactactgttggttctgctgctgaaggttcactagataaaaga BBa_J63003_sequence 1 cccgccgccaccatggag BBa_K392003_sequence 1 gcatgtgctctgtatgtatataaaactcttgttttcttcttttctctaaatattctttccttatacattaggacctttgcagcataaattactatacttctattactagagcccgccgccaccatggag BBa_K1772003_sequence 1 atgtcatcacctgatccggctagtgatcagttggtgaattacaaaaaaaaacaaacagataaaactaagatagtaactgggcatggtgctccagttgacaataggggagcgagtttaaccgtgggtccgagaggtcctatgctgttgcaagacattacctttctgaacgaattggctcacttcgatcgtgaaaggataccagaaagggtcgtacatgctaagggagccggcgcttttggttattttgaagttacgcatgatattactaaatattgtaaagcctcagttttcagcaagattggaaaaaaaacgcccattgctgtaagattctcaacagtcggtggtgaatctgggagtgcggatactgtacgtgatcccagaggatttgcggttaaattttacactgaagatgggatatgggacctggttgggaataatacccctattttctttatcagagatcctttattgttccccgtatttattcacacccaaaagagaaacccggcaacacacctgaaagactgtgatatgttttgggatttcttgagcctaagacccgagtcaacacatcaagtaatgtttttgttttctgataggggtataccggatggctttagacatatgaacggctatggttctcatacttttaaggccataaatgataaaaacgaagcagtatacgtaaagttccactacaagacaaatcaaggcataaaaaacttattagcacagaaagcttctgaagtcgcagtagcagatccggattattctataagagacttatacaatgctattgcgagaggtcaatatccatcctggactctgtatatacaggtcatgacatttgaacaagctgaaaagtttagatggaacccctttgatctaaccaaagtatggcctcatgccgaataccccctaattccagttggaaaattggtcctagataggaaccctgccaattattttgctgaagtagaacaaatcgcattttccccagcgcatatggttcccggaattgaaccaagtccggacaaaatgttgcaaggtagattattcagttattctgatacacataggcacagattaggggccaattatctacagattccggtaaattgcccctatcgtactagaattacaaattatcaaagagacggtcctcaaacttttaccaacaatcaagaaggagctccaaactattatcctaattctttttctggtcctgaagacgttcctcattgtgctgccattaagtttgctagcactggtgacgttgcaagatacaattcaggtgatgaagacaatttcagtcaacctagcttattttggaagaagacacttaaacctgaggagagagaacgtttggtgcaaaatattgtcgatcatgtgaaagatgctgcggattttgtgcaggaaagaaccgtcaaaaatttctctcaagtggatgctgaattcggcaggaaactgactgagggtctaagaaagcactcaaagaattcaagcatcgcttccgccaaccta BBa_K1772004_sequence 1 cagatccgccaggcgtgtatatatagcgtggatggccaggcaactttagtgctgacacatacaggcatatatatatgtgtgcgacgacacatgatcatatggcatgcatgtgctctgtatgtatataaaactcttgttttcttcttttctctaaatattctttccttatacattaggacctttgcagcataaattactatacttctatagacacacaaacacaaatacacacactaaattaatatactagagcccgccgccaccatggagtactagagggtggttctggttactagatgtcatcacctgatccggctagtgatcagttggtgaattacaaaaaaaaacaaacagataaaactaagatagtaactgggcatggtgctccagttgacaataggggagcgagtttaaccgtgggtccgagaggtcctatgctgttgcaagacattacctttctgaacgaattggctcacttcgatcgtgaaaggataccagaaagggtcgtacatgctaagggagccggcgcttttggttattttgaagttacgcatgatattactaaatattgtaaagcctcagttttcagcaagattggaaaaaaaacgcccattgctgtaagattctcaacagtcggtggtgaatctgggagtgcggatactgtacgtgatcccagaggatttgcggttaaattttacactgaagatgggatatgggacctggttgggaataatacccctattttctttatcagagatcctttattgttccccgtatttattcacacccaaaagagaaacccggcaacacacctgaaagactgtgatatgttttgggatttcttgagcctaagacccgagtcaacacatcaagtaatgtttttgttttctgataggggtataccggatggctttagacatatgaacggctatggttctcatacttttaaggccataaatgataaaaacgaagcagtatacgtaaagttccactacaagacaaatcaaggcataaaaaacttattagcacagaaagcttctgaagtcgcagtagcagatccggattattctataagagacttatacaatgctattgcgagaggtcaatatccatcctggactctgtatatacaggtcatgacatttgaacaagctgaaaagtttagatggaacccctttgatctaaccaaagtatggcctcatgccgaataccccctaattccagttggaaaattggtcctagataggaaccctgccaattattttgctgaagtagaacaaatcgcattttccccagcgcatatggttcccggaattgaaccaagtccggacaaaatgttgcaaggtagattattcagttattctgatacacataggcacagattaggggccaattatctacagattccggtaaattgcccctatcgtactagaattacaaattatcaaagagacggtcctcaaacttttaccaacaatcaagaaggagctccaaactattatcctaattctttttctggtcctgaagacgttcctcattgtgctgccattaagtttgctagcactggtgacgttgcaagatacaattcaggtgatgaagacaatttcagtcaacctagcttattttggaagaagacacttaaacctgaggagagagaacgtttggtgcaaaatattgtcgatcatgtgaaagatgctgcggattttgtgcaggaaagaaccgtcaaaaatttctctcaagtggatgctgaattcggcaggaaactgactgagggtctaagaaagcactcaaagaattcaagcatcgcttccgccaacctatactagagggtggttctggttactagatgaaagttttgattgttttgttggctattttcgctgctttgccattggctttggctcaaccagttatttctactactgttggttctgctgctgaaggttcactagataaaagatactagaggcatgtgctctgtatgtatataaaactcttgttttcttcttttctctaaatattctttccttatacattaggacctttgcagcataaattactatacttctattactagagcccgccgccaccatggag igem2sbol 1 iGEM to SBOL conversion Conversion of the iGEM parts registry to SBOL2.1 James Alastair McLaughlin Chris J. Myers 2017-03-06T15:00:00.000Z