BBa_Y00100 1 BBa_Y00100 nuclear 5-aminolevulinate synthase gene from S. cerevisiae, recoded for mitochondrial expression 2006-08-03T11:00:00Z 2015-05-08T01:14:54Z direct synthesis from DNA 2.0, summer 2006 The first step in S. cerevisiae's pathway for heme biosynthesis is catalyzed by the product of the HEM1 gene [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=HEM1]. HEM1 is a nuclear gene (1.647 kb,[http://db.yeastgenome.org/cgi-bin/getSeq?seq=YDR232W&flankl=0&flankr=0&map=a3map]) that encodes the enzyme 5-aminolevulinate ("ALA" or "d-ALA") synthase (549 aa,[http://db.yeastgenome.org/cgi-bin/getSeq?seq=YDR232W&flankl=0&flankr=0&map=p3map]). The enzyme is post-translationally imported into the mitochondria where it is responsible for converting glycine and succinyl-CoA to 5-aminolevulinate ("d-ALA") plus CoA and CO2. Deletion of HEM1 is lethal for the cell but viabilty can be restored by providing the cells with the product of the HEM1p catalyzed reaction, namely d-ALA. In an effort to directly express the HEM1 gene from the mitochondrial genome, the HEM1 gene was recoded by DNA 2.0 for mitochondrial codons, an epitope tag (HA = YPYDVPDYA) was added to the C-terminal portion of the protein just before the stop codon, and flanking sequences (100 bp upstm of ATG and 92 dwstm of stop) were added to facilitate integration into mitochondrial genome. false false _43_ 0 314 1 It's complicated false codon optimized for mitochondrial gene expression, HA-epitope C term (not N-term where mt import sequences rest), flank coding seq with homology to COX3 gene of mitochondria to allow for integration and disruption of this gene (should make respiration+ cells become respiration-), and finally added BB prefix and suffix to allow cloning into std Registry vectors false Natalie Kuldell annotation1975604 1 for recombination to COX3 range1975604 1 1775 1865 annotation1975601 1 for recombination to COX3 range1975601 1 1 101 annotation1975602 1 HA range1975602 1 1745 1771 annotation1975605 1 start codon range1975605 1 101 103 annotation1975603 1 TGA stop codon range1975603 1 1772 1774 annotation1975606 1 coding sequence range1975606 1 101 1774 BBa_Y00100_sequence 1 aattttattatatagtttaagggataatattttattaatattttttttatttatttatttaattatattatatatataatatatatataacaataaatttatgcaacgttctatattcgccagatttggtaacagtagtgctgctgtgtccactttaaacagactgtctacgacggctgcgcctcatgctaaaaacggctacgcaactgctactggcgccggtgcagcagcagctacagcaaccgcatcctctacacatgcagctgccgccgcagccgccgctgccaaccattctacacaagaatctggcttcgattatgagggactaattgattcagaattgcagaaaaagcgtttggacaaatcttatagatactttaacaacattaacaggctggccaaagagttcccattagcccatagacagagagaagccgataaggttacggtctggtgctcaaatgattatcttgctttatccaaacatccagaggtgcttgatgctatgcacaaaacaatcgataaatacggatgcggtgctggtggtactcgtaacattgccggacataacataccaaccttgaacttagaagcagaattggctacgttacacaaaaaggagggtgctttggtattttcttcatgttatgtggccaatgatgcagttctgagcttattgggccagaagatgaaagatctagtgatcttcagcgatgaactaaaccacgcctccatgattgttgggatcaaacatgcaaatgttaagaaacacatttttaagcacaatgatctgaatgagcttgaacaactgttacaatcatatcccaaatcagtacctaaattgattgcttttgaatctgtgtattcgatggcgggcagtgttgcggatatagaaaagatttgcgaccttgccgataagtatggtgctctaacctttttagatgaagttcatgccgttggtttgtacggccctcacggcgctggtgttgccgaacattgtgactttgaatcacatagagcttccggtattgcgacgccaaaaactaatgataagggtggtgctaaaactgtcatggatagagtggatatgattactggtacattagggaaatcctttggctctgtaggaggatatgttgcggcgtcaagaaaacttattgactggtttagatctttcgccccaggtttcatttttacaacaactttgcccccatcagtgatggcaggggccacagccgccataagatatcaaaggtgtcatattgatctacgtacatctcaacaaaagcacactatgtacgtcaaaaaggcatttcatgagctgggtattcctgtgatcccgaatccttctcatattgtacctgtgttgataggtaacgctgatctagcaaaacaagcctccgatattctgatcaacaagcatcaaatttatgtccaagcaatcaactttccaacagttgctagaggaactgaaaggctgagaataacgcctactccaggtcacactaacgatttgagtgatattttgattaatgccgtggatgatgtctttaacgaattgcaattacccagagttagagactgggagagtcaaggtggtcttctgggggtgggagaaagcggctttgttgaagaatctaatttatggacgagcagtcaattatccttgactaacgatgacctgaatcctaatgtcagagatccgatcgtaaagcaattagaagtttcgtctgggataaaacagtatccatacgatgtacccgattacgcgtgaaattatatatctaaatgattaatatatatattattaataattaacaataattaatatattataatttatatatatatattttatattattat 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