LOCUS Exported 6742 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION LacI-Mit1AD on pPICZ B expression vector. ACCESSION . VERSION . KEYWORDS pZ_P-ICL1-LacI-Mit1AD SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6742) TITLE Engineered ethanol-driven biosynthetic system for improving production of acetyl-CoA derived drugs in Crabtree-negative yeast REFERENCE 2 (bases 1 to 6742) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6742 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 4130..4147 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" terminator 4227..4473 /gene="Pichia pastoris AOX1" /label=AOX1 terminator /note="transcription terminator for AOX1" promoter 4494..4901 /gene="S. cerevisiae TEF1" /label=TEF1 promoter /note="promoter for EF-1-alpha" promoter 4908..4955 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 4974..5348 /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 5414..5661 /gene="S. cerevisiae CYC1" /label=CYC1 terminator /note="transcription terminator for CYC1" primer_bind complement(5430..5448) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" rep_origin complement(5736..6324) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5816..5835) /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 caaatgtgct gttccggtag cttgtaggaa gcggcatccg tagggcaata tacgactata 61 gcttctaaag cgtagtacaa tgaaatgttc gaaggaacaa caaacggatt tgtttttcgt 121 aggctcaacc cgttgaggtg taactcttta gcgaaagggt aagattgatt gttcgaagta 181 gggcctcaaa gggaaagaga aaaaaaaaaa tacaccaaga gttacgtaag catatatttt 241 ttacgtaaag catgattgaa tttcagcagt attgtttaac aaggctgatg tcgtgtgcca 301 atcaaaacaa aagagattcg cataatgcca taattggggt gtgtgggcgc cccctaaaac 361 gtctttctca tcatcatctg caacccccat cgaacctcat taaatcacat gacttgtgcg 421 atcctcggtc aactcgttcc gtgcacccat tccaccccgg gctgaccaac gcaaggttct 481 ccgagagtcc gctaccccag atttatatca gcaaccagtc acctttttcc gggcacgact 541 ctatatgccc tggaaaaccg gagacgatga gcctgactgt aaaaggtgac agaaccccca 601 actctggtta atctcttcaa caaatacttt attttctttc aattcaaaga acacagtatc 661 aagtatatca agaatgggtg ttaagccagt tactttgtat gacgttgctg aatacgctgg 721 agtttcctac caaactgtct ctagagttgt taatcaagct tctcatgtct ccgctaagac 781 tagagagaag gttgaggctg ctatggctga attgaactat attccaaata gagttgctca 841 gcagttggct ggaaagcaat ctttgttgat tggagtcgct acttcttctt tggctttgca 901 tgctccatct cagattgttg ctgctattaa gtccagagct gaccagttgg gagcttctgt 961 tgttgtttct atggttgaga gatctggagt tgaggcttgc aaggctgctg ttcataactt 1021 gttggctcag agagtttctg gattgattat taattaccca ttggacgatc aagacgctat 1081 tgccgttgag gccgcttgta ccaacgtccc agctttgttc ttggacgttt ccgatcaaac 1141 tccaattaat tctattattt tttctcacga ggatggaact agattgggag ttgaacactt 1201 ggttgctttg ggacatcaac agattgcttt gttggctgga ccattgtctt ccgtttctgc 1261 tagattgaga ttggccggat ggcacaagta cttgaccaga aaccagattc aaccaattgc 1321 tgagagagag ggagattggt ctgctatgtc tggattccag cagactatgc agatgttgaa 1381 cgaaggaatt gtcccaaccg ctatgttggt cgctaatgac caaatggctt tgggagctat 1441 gagagctatt actgaatctg gattgagagt cggagctgac atttctgttg ttggatatga 1501 tgacactgag gattcttctt gctacattcc accattgact actattaagc aagacttcag 1561 attgttggga cagacttctg ttgatagatt gttgcagttg tcccaaggac aagctgttaa 1621 aggaaaccaa ttgttgccag tttctttggt taagagaaag actactttgg ctccaaacac 1681 tcagactgct tccccaagag ctttggctga ctctttgatg caattggcta gacaagtctc 1741 tagattggag tctggacaag gtggcggcgg ctctgttaac aactccatga aggatttctt 1801 aggcaagaaa acggtggatg gagctgatag tctcaatttg gccgtgaatc tgcaacaaca 1861 gcagagttca aacacaattg ccaatcaatc gctttcctca attggattgg aaagttttgg 1921 ttacggctct ggtatcaaaa acgagtttaa cttccaagac ttgataggtt caaactctgg 1981 cagttcagat ccgacatttt cagtagacgc tgacgaggcc caaaaactcg acatttccaa 2041 caagaacagt cgtaagagac agaaactagg tttgctgccg gtcagcaatg caacttccca 2101 tttgaacggt ttcaatggaa tgtccaatgg aaagtcacac tctttctctt caccgtctgg 2161 gactaatgac gatgaactaa gtggcttgat gttcaactca ccaagcttca accccctcac 2221 agttaacgat tctaccaaca acagcaacca caatataggt ttgtctccga tgtcatgctt 2281 attttctaca gttcaagaag catctcaaaa aaagcatgga aattccagta gacacttttc 2341 atacccatct gggccggagg acctttggtt caatgagttc caaaaacagg ccctcacagc 2401 caatggagaa aatgctgtcc aacagggaga tgatgcttct aagaacaaca cagccattcc 2461 taaggaccag tcttcgaact catcgatttt cagttcacgt tctagtgcag cttctagcaa 2521 ctcaggagac gatattggaa ggatgggccc attctccaaa ggaccagaga ttgagttcaa 2581 ctacgattct tttttggaat cgttgaaggc agagtcaccc tcttcttcaa agtacaatct 2641 gccggaaact ttgaaagagt acatgaccct tagttcgtct catctgaata gtcaacactc 2701 cgacactttg gcaaatggca ctaacggtaa ctattctagc accgtttcca acaacttgag 2761 cttaagtttg aactccttct ctttctctga caagttctca ttgagtccac caacaatcac 2821 tgacgccgaa aagttttcat tgatgagaaa cttcattgac aacatctcgc catggtttga 2881 cacttttgac aataccaaac agtttggaac aaaaattcca gttctggcca aaaaatgttc 2941 ttcattgtac tatgccattc tggctatatc ttctcgtcaa agagaaagga taaagaaaga 3001 gcacaatgaa aaaacattgc aatgctacca atactcacta caacagctca tccctactgt 3061 tcaaagctca aataatattg agtacattat cacatgtatt ctcctgagtg tgttccacat 3121 catgtctagt gaaccttcaa cccagaggga catcattgtg tcattggcaa aatacattca 3181 agcatgcaac ataaacggat ttacatctaa tgacaaactg gaaaagagta ttttctggaa 3241 ctatgtcaat ttggatttgg ctacttgtgc aatcggtgaa gagtcaatgg tcattccttt 3301 tagctactgg gttaaagaga caactgacta caagaccatt caagatgtga agccattttt 3361 caccaagaag actagcacga caactgacga tgacttggac gatatgtatg ccatctacat 3421 gctgtacatt agtggtagaa tcattaacct gttgaactgc agagatgcga agctcaattt 3481 tgagcccaag tgggagtttt tgtggaatga actcaatgaa tgggaattga acaaaccctt 3541 gacctttcaa agtattgttc agttcaaggc caatgacgaa tcgcagggcg gatcaacttt 3601 tccaactgtt ctattctcca actctcgaag ctgttacagt aaccagctgt atcatatgag 3661 ctacatcatc ttagtgcaga ataaaccacg attatacaaa atccccttta ctacagtttc 3721 tgcttcaatg tcatctccat cggacaacaa agctgggatg tctgcttcca gcacacctgc 3781 ttcagaccac cacgcttctg gtgatcattt gtctccaaga agtgtagagc cctctctttc 3841 gacaacgttg agccctccgc ctaatgcaaa cggtgcaggt aacaagttcc gctctacgct 3901 ctggcatgcc aagcagatct gtgggatttc tatcaacaac aaccacaaca gcaatctagc 3961 agccaaagtg aactcattgc aaccattgtg gcacgctgga aagctaatta gttccaagtc 4021 tgaacataca cagttgctga aactgttgaa caaccttgag tgtgcaacag gctggcctat 4081 gaactggaag ggcaaggagt taattgacta ctggaatgtt gaagaataac atcatcatca 4141 tcatcattga gtttgtagcc ttagacatga ctgttcctca gttcaagttg ggcacttacg 4201 agaagaccgg tcttgctaga ttctaatcaa gaggatgtca gaatgccatt tgcctgagag 4261 atgcaggctt catttttgat acttttttat ttgtaaccta tatagtatag gatttttttt 4321 gtcattttgt ttcttctcgt acgagcttgc tcctgatcag cctatctcgc agctgatgaa 4381 tatcttgtgg taggggtttg ggaaaatcat tcgagtttga tgtttttctt ggtatttccc 4441 actcctcttc agagtacaga agattaagtg agaccttcgt ttgtgcggat cccccacaca 4501 ccatagcttc aaaatgtttc tactcctttt ttactcttcc agattttctc ggactccgcg 4561 catcgccgta ccacttcaaa acacccaagc acagcatact aaatttcccc tctttcttcc 4621 tctagggtgt cgttaattac ccgtactaaa ggtttggaaa agaaaaaaga gaccgcctcg 4681 tttctttttc ttcgtcgaaa aaggcaataa aaatttttat cacgtttctt tttcttgaaa 4741 attttttttt ttgatttttt tctctttcga tgacctccca ttgatattta agttaataaa 4801 cggtcttcaa tttctcaagt ttcagtttca tttttcttgt tctattacaa ctttttttac 4861 ttcttgctca ttagaaagaa agcatagcaa tctaatctaa gggcggtgtt gacaattaat 4921 catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca 4981 agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct 5041 ggaccgaccg gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc 5101 gggacgacgt gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc 5161 tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt 5221 ccacgaactt ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg 5281 ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc 5341 aggactgaca cgtccgacgc ggcccgacgg gtccgaggcc tcggagatcc gtcccccttt 5401 tcctttgtcg atatcatgta attagttatg tcacgcttac attcacgccc tccccccaca 5461 tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt 5521 tttatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg 5581 tacagacgcg tgtacgcatg taacattata ctgaaaacct tgcttgagaa ggttttggga 5641 cgctcgaagg ctttaatttg caagctggag accaacatgt gagcaaaagg ccagcaaaag 5701 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 5761 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 5821 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 5881 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc 5941 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 6001 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 6061 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 6121 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca 6181 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 6241 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 6301 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 6361 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gatcagatct tcatctaaca 6421 ctttgtatag cacatcgtac cagtttatca aaatccaaca agttttcctc tgggtactga 6481 aattcgatta gtaataaacg ggcgcacttg gtgatctcct catcctgttc actagccttc 6541 ttttcgctta acttcatcaa cgagatcaac tcgagtctgc gagcagagaa ctgtttccaa 6601 agagtttcat cgctgaaatt cagcttcttt cgaataaaat gtgtcactcc cactttatta 6661 ctcgtggtgt tatttgtttt ccacgaacta gttgaatcgc catctttact accgtcctgg 6721 gtgacatccg gcgaattcgg ga //