LOCUS Exported 3584 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION yeast plasmid overexpressing LovC. ACCESSION . VERSION . KEYWORDS pZ_lacO-cP-AOX1-LovC SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3584) TITLE Engineered ethanol-driven biosynthetic system for improving production of acetyl-CoA derived drugs in Crabtree-negative yeast REFERENCE 2 (bases 1 to 3584) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3584 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 64..80 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." CDS 1358..1375 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" terminator 1455..1701 /gene="Pichia pastoris AOX1" /label=AOX1 terminator /note="transcription terminator for AOX1" promoter 1722..2129 /gene="S. cerevisiae TEF1" /label=TEF1 promoter /note="promoter for EF-1-alpha" promoter 2136..2183 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 2202..2576 /codon_start=1 /gene="Sh ble from Streptoalloteichus hindustanus" /product="antibiotic-binding protein" /label=BleoR /note="confers resistance to bleomycin, phleomycin, and Zeocin(TM)" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" terminator 2642..2889 /gene="S. cerevisiae CYC1" /label=CYC1 terminator /note="transcription terminator for CYC1" primer_bind complement(2658..2676) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" rep_origin complement(2964..3552) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3044..3063) /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagatcag atcttgtgtg 61 gaattgtgag cggataacaa tttcacacac taacccctac ttgacagcaa tatataaaca 121 gaaggaagct gccctgtctt aaaccttttt ttttatcatc attattagct tactttcata 181 attgcgactg gttccaattg acaagctttt gattttaacg acttttaacg acaacttgag 241 aagatcaaaa aacaactaat tattcgaaat gggcgaccag ccattcattc caccaccgca 301 gcaaacagcg ctgacggtaa atgaccatga tgaagtcacc gtctggaatg ccgcaccctg 361 ccccatgctg ccccgcgacc aggtatacgt ccgcgtcgag gccgtggcga tcaatcccag 421 tgacacgaag atgcgcggac agtttgccac gccctgggcg tttctcggaa cggactatgc 481 cggcacggtc gtcgcagtgg gttcggacgt gactcatatc caagtgggtg accgggtcta 541 cggggcacag aacgagatgt gcccacgcac cccggatcag ggggcattct cgcagtacac 601 ggtcacgcga ggccgtgttt gggccaagat ccccaagggc ttgtcgttcg agcaggctgc 661 cgcgctacct gcgggcatca gtaccgctgg attggcgatg aagttgcttg ggctgccttt 721 gccatcgcct tcggcagacc agccacccac ccactccaag ccggtgtatg tgttggtcta 781 tgggggcagt acggccactg ccactgtcac tatgcaaatg ctccgcctgt ccggatatat 841 tccaattgca acatgctccc cccacaattt cgacctggcc aaatcgcgcg gcgcagagga 901 ggtctttgac tatcgggccc cgaatctcgc gcagacgatc cgtacctaca ccaagaacaa 961 tctccgctat gctctcgact gtatcaccaa cgtcgagtcc accacattct gcttcgcagc 1021 catcggccgc gcgggggggc actacgtctc cctgaacccg ttccctgaac acgcggccac 1081 gcgcaagatg gtcacgaccg actggaccct ggggccgacc atctttggcg agggatcaac 1141 ctggcccgcc ccctatgggc gtcccggcag tgaggaagag cggcagttcg gcgaggatct 1201 gtggcgcatc gcggggcagc tcgtcgaaga tggacgcctc gtccatcatc cgttgcgcgt 1261 ggtgcagggc ggcttcgatc acattaagca aggcatggag ctcgtccgga agggagagct 1321 gtcgggggag aaactcgtgg ttcggctcga ggggccgcat catcatcatc atcattgagt 1381 ttgtagcctt agacatgact gttcctcagt tcaagttggg cacttacgag aagaccggtc 1441 ttgctagatt ctaatcaaga ggatgtcaga atgccatttg cctgagagat gcaggcttca 1501 tttttgatac ttttttattt gtaacctata tagtatagga ttttttttgt cattttgttt 1561 cttctcgtac gagcttgctc ctgatcagcc tatctcgcag ctgatgaata tcttgtggta 1621 ggggtttggg aaaatcattc gagtttgatg tttttcttgg tatttcccac tcctcttcag 1681 agtacagaag attaagtgag accttcgttt gtgcggatcc cccacacacc atagcttcaa 1741 aatgtttcta ctcctttttt actcttccag attttctcgg actccgcgca tcgccgtacc 1801 acttcaaaac acccaagcac agcatactaa atttcccctc tttcttcctc tagggtgtcg 1861 ttaattaccc gtactaaagg tttggaaaag aaaaaagaga ccgcctcgtt tctttttctt 1921 cgtcgaaaaa ggcaataaaa atttttatca cgtttctttt tcttgaaaat tttttttttt 1981 gatttttttc tctttcgatg acctcccatt gatatttaag ttaataaacg gtcttcaatt 2041 tctcaagttt cagtttcatt tttcttgttc tattacaact ttttttactt cttgctcatt 2101 agaaagaaag catagcaatc taatctaagg gcggtgttga caattaatca tcggcatagt 2161 atatcggcat agtataatac gacaaggtga ggaactaaac catggccaag ttgaccagtg 2221 ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg accgaccggc 2281 tcgggttctc ccgggacttc gtggaggacg acttcgccgg tgtggtccgg gacgacgtga 2341 ccctgttcat cagcgcggtc caggaccagg tggtgccgga caacaccctg gcctgggtgt 2401 gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc 2461 gggacgcctc cgggccggcc atgaccgaga tcggcgagca gccgtggggg cgggagttcg 2521 ccctgcgcga cccggccggc aactgcgtgc acttcgtggc cgaggagcag gactgacacg 2581 tccgacgcgg cccgacgggt ccgaggcctc ggagatccgt cccccttttc ctttgtcgat 2641 atcatgtaat tagttatgtc acgcttacat tcacgccctc cccccacatc cgctctaacc 2701 gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt tatagttatg 2761 ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta cagacgcgtg 2821 tacgcatgta acattatact gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct 2881 ttaatttgca agctggagac caacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 2941 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 3001 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 3061 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 3121 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc 3181 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 3241 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 3301 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 3361 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc 3421 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 3481 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 3541 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctga //