LOCUS       40924_46343        8783 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pWCD2139, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8783)
  AUTHORS   DeLoache WC, Russ ZN, Narcross L, Gonzales AM, Martin VJ, Dueber JE.
  TITLE     An enzyme-coupled biosensor enables (S)-reticuline production in 
            yeast from glucose
  JOURNAL   Nat. Chem. Biol. 11 (7), 465-471 (2015)
  PUBMED    25984720
REFERENCE   2  (bases 1 to 8783)
  AUTHORS   DeLoache WC, Russ ZN, Narcross L, Gonzales AM, Martin VJJ., Dueber 
            JE.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-APR-2015) Department of Bioengineering, University of 
            California, Berkeley, 2151 Berkeley Way Room 542, Berkeley, CA 
            94704, USA
REFERENCE   3  (bases 1 to 8783)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8783)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. Chem. 
            Biol."; date: "2015"; volume: "11"; issue: "7"; pages: "465-471"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (22-APR-2015) Department of Bioengineering, University of 
            California, Berkeley, 2151 Berkeley Way Room 542, Berkeley, CA 
            94704, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8783
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    5..524
                     /label=similar to URA3 5'
                     /note="similar to URA3 5'"
     promoter        586..1252
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate
                     dehydrogenase; also known as the TDH3 promoter"
     CDS             1259..2749
                     /gene="CYP76AD1"
                     /label=CYP76AD1
                     /note="Cytochrome P450 76AD1 from Beta vulgaris.
                     Accession#: I3PFJ5"
     regulatory      2765..2988
                     /regulatory_class="terminator"
     regulatory      3022..3721
                     /regulatory_class="promoter"
     CDS             3728..5146
                     /codon_start=1
                     /product="DOPA decarboxylase"
                     /label=DOPA decarboxylase
                     /note="PpDODC (DOPA decarboxylase) derived from Pseudomonas
                     putida"
                     /protein_id="AKH61544.1"
                     /translation="MTPEQFRQYGHQLIDLIADYRQTVGERPVMAQVEPGYLKAALPAT
                     APQQGEPFAAILDDVNNLVMPGLSHWQHPDFYGYFPSNGTLSSVLGDFLSTGLGVLGLS
                     WQSSPALSELEETTLDWLRQLLGLSGQWSGVIQDTASTSTLVALISARERATDYALVRG
                     GLQAEPKPLIVYVSAHAHSSVDKAALLAGFGRDNIRLIPTDERYALRPEALQAAIEQDL
                     AAGNQPCAVVATTGTTTTTALDPLRPVGEIAQANGLWLHVDSAMAGSAMILPECRWMWD
                     GIELADSVVVNAHKWLGVAFDCSIYYVRDPQHLIRVMSTNPSYLQSAVDGEVKNLRDWG
                     IPLGRRFRALKLWFMLRSEGVDALQARLRRDLDNAQWLAGQVEAAAEWEVLAPVQLQTL
                     CIRHRPAGLEGEALDAHTKGWAERLNASGAAYVTPATLDGRWMVRVSIGALPTERGDVQ
                     RLWARLQDVIKGGS"
     terminator      5153..5340
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     regulatory      5426..5602
                     /regulatory_class="promoter"
     CDS             5603..6403
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     regulatory      6407..6484
                     /regulatory_class="terminator"
     CDS             6674..6934
                     /gene="TIM9"
                     /label=TIM9
                     /note="Mitochondrial import inner membrane translocase
                     subunit TIM9 from Saccharomyces cerevisiae (strain ATCC 
                     204508 / S288c). Accession#: O74700"
     regulatory      complement(7021..7047)
                     /regulatory_class="terminator"
     CDS             complement(7051..7863)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     regulatory      complement(7864..8011)
                     /regulatory_class="promoter"
     rep_origin      8088..8676
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 caatgctaaa ttcgagtgaa acacaggaag atcagaaaat cctcatttca tccatattaa
       61 caataatttc aaatgtttat ttgcattatt tgaaactagg caagacaagc aacgaaacgt
      121 ttttgaaaat tttgagtatt ttcaataaat ttgtagagga ctcagatatt gaaaaaaagc
      181 tacagcaatt aatacttgat aagaagagta ttgagaaggg caacggttca tcatctcatg
      241 gatctgcaca tgaacaaaca ccagagtcaa acgacgttga aattgaggct actgcgccaa
      301 ttgatgacaa tacagacgat gataacaaac cgaagttatc tgatgtagaa aaggattaaa
      361 gatgctaaga gatagtgatg atatttcata aataatgtaa ttctatatat gttaattacc
      421 ttttttgcga ggcatattta tggtgaagga taagttttga ccatcaaaga aggttaatgt
      481 ggctgtggtt tcagggtcca taaagcccac atggataaca ttacccctga attcgcatct
      541 agactgaact ggccgataat tgcagacgaa cgcagttcga gtttatcatt atcaatactg
      601 ccatttcaaa gaatacgtaa ataattaata gtagtgattt tcctaacttt atttagtcaa
      661 aaaattagcc ttttaattct gctgtaaccc gtacatgccc aaaatagggg gcgggttaca
      721 cagaatatat aacatcgtag gtgtctgggt gaacagttta ttcctggcat ccactaaata
      781 taatggagcc cgctttttaa gctggcatcc agaaaaaaaa agaatcccag caccaaaata
      841 ttgttttctt caccaaccat cagttcatag gtccattctc ttagcgcaac tacagagaac
      901 aggggcacaa acaggcaaaa aacgggcaca acctcaatgg agtgatgcaa cctgcctgga
      961 gtaaatgatg acacaaggca attgacccac gcatgtatct atctcatttt cttacacctt
     1021 ctattacctt ctgctctctc tgatttggaa aaagctgaaa aaaaaggttg aaaccagttc
     1081 cctgaaatta ttcccctact tgactaataa gtatataaag acggtaggta ttgattgtaa
     1141 ttctgtaaat ctatttctta aacttcttaa attctacttt tatagttagt ctttttttta
     1201 gttttaaaac accaagaact tagtttcgaa taaacacaca taaacaaaca aaagatctat
     1261 ggatcatgca acattagcta tgatattagc aatttggttt atttcatttc attttattaa
     1321 attattattc tcacaacaaa caacaaaatt gttacctcca ggtcctaaac ctttaccaat
     1381 tattggtaat atcttggaag taggaaagaa gcctcataga tcatttgcaa atttggctaa
     1441 gatccacggt cctttaatct ctttgaggtt aggatcagtt accaccattg ttgtatcatc
     1501 tgctgatgtc gccaaagaga tgttcttgaa gaaggatcac cctttatcaa acaggacaat
     1561 tccaaattca gtaactgctg gtgaccacca caaattgact atgtcttggt tgcctgtttc
     1621 acctaaatgg agaaacttca gaaagataac tgctgttcat ttattgtctc cacaaagatt
     1681 agatgcatgc cagaccttta gacatgcaaa ggttcagcaa ttatatgaat acgttcaaga
     1741 gtgcgctcag aagggtcagg cagttgacat tggtaaagct gccttcacaa cttctttgaa
     1801 tttattgtct aaattattct tttctgttga attggcccat cataaatcac atacctctca
     1861 agagtttaag gaattaattt ggaatattat ggaagacatc ggtaaaccaa actacgccga
     1921 ttacttccca attttaggtt gcgtcgatcc atcaggaatt agaagaaggt tggcatgttc
     1981 ttttgataag ttaattgcag tatttcaagg aataatatgt gagaggttag ctccagattc
     2041 atcaactaca actactacaa ctacagatga tgtcttggac gtattattgc aattgtttaa
     2101 gcagaatgaa ttaactatgg gagagattaa ccacttgtta gtcgatattt tcgatgccgg
     2161 tactgatact acatcatcta ccttagagtg ggttatgact gagttaataa gaaaccctga
     2221 aatgatggaa aaggcccaag aggaaataaa acaggtattg ggtaaggata agcaaatcca
     2281 agagtcagac ataataaatt tgccatactt gcaagcaatc attaaggaaa ctttgaggtt
     2341 gcacccacct acagttttct tgttgcctag aaaagccgac acagacgtag aattgtacgg
     2401 atacattgtt ccaaaggatg ctcaaatttt ggtaaacttg tgggccatag gtagagatcc
     2461 aaatgcttgg caaaatgccg atattttctc acctgaaaga tttataggtt gtgagattga
     2521 tgtaaaggga agagattttg gtttattgcc ttttggtgct ggtagaagaa tttgccctgg
     2581 tatgaatttg gccataagga tgttgacatt gatgttggcc acattattgc aattcttcaa
     2641 ctggaaatta gagggagaca tttcacctaa agatttggac atggatgaaa agtttggtat
     2701 agccttgcag aaaactaaac ctttaaaatt gatcccaatt ccaagatacg gatcctaact
     2761 cgagataaag caatcttgat gaggataatg attttttttt gaatatacat aaatactacc
     2821 gtttttctgc tagattttgt gatgacgtaa ataagtacat attacttttt aagccaagac
     2881 aagattaagc attaacttta cccttttctt tctaagtttc aatattagtt atcactgttt
     2941 aaaagttatg gcgagaacgt cggcggttaa aatatattac cctgaacggc tgccaaacca
     3001 gatgtcaaca cagctacaac gcacccatga accacacggt tagtccaaaa ggggcagttc
     3061 agattccaga tgcgggaatt agcttgctgc caccctcacc tcactaacgc tgcggtgtgc
     3121 ggatacttca tgctatttat agacgcgcgt gtcggaatca gcacgcgcaa gaaccaaatg
     3181 ggaaaatcgg aatgggtcca gaactgcttt gagtgctggc tattggcgtc tgatttccgt
     3241 tttgggaatc ctttgccgcg cgcccctctc aaaactccgc acaagtccca gaaagcggga
     3301 aagaaataaa acgccaccaa aaaaaaaaaa ataaaagcca atcctcgaag cgtgggtggt
     3361 aggccctgga ttatcccgta caagtatttc tcaggagtaa aaaaaccgtt tgttttggaa
     3421 tttcccattt cgcggccacc tacgccgcta tctttgcaac aactatctgc gataactcag
     3481 caaattttgc atattcgtgt tgcagtattg cgataatggg agtcttactt ccaacataac
     3541 ggcagaaaga aatgtgagaa aattttgcat cctttgcctc cgttcaagta tataaagtcg
     3601 gcatgcttga taatctttct ttccatccta cattgttcta attattctta ttctccttta
     3661 ttctttccta acataccaag aaattaatct tctgtcattc gcttaaacac tatatcaata
     3721 aagatctatg actcctgaac aatttaggca atatggtcat caattaatcg atttgatcgc
     3781 cgattacaga cagaccgttg gagaaagacc tgtaatggct caagttgaac ctggatattt
     3841 aaaggccgct ttgcctgcta ccgcaccaca acaaggtgaa ccattcgctg ccattttgga
     3901 tgatgtaaac aatttggtca tgccaggatt gtcacattgg caacacccag acttttatgg
     3961 ttatttccct tcaaatggaa cattatcatc tgttttgggt gacttcttat ctacaggttt
     4021 aggagtattg ggtttatctt ggcaatcttc tccagcattg tctgaattgg aagaaaccac
     4081 cttagactgg ttaagacaat tgttgggttt atctggacag tggtctggag ttatacaaga
     4141 caccgcatct acctcaactt tggtcgcatt gatttcagct agagagagag caacagatta
     4201 cgcattggtc aggggtggtt tgcaagccga accaaaacca ttgattgtat atgtctctgc
     4261 acacgctcat tcatcagttg acaaagctgc attattggct ggtttcggta gagataacat
     4321 aagattaatc ccaacagacg agaggtatgc attaagacct gaagcattac aagctgcaat
     4381 tgaacaggac ttggcagctg gaaaccagcc ttgtgctgtt gtcgctacta ctggtacaac
     4441 tacaactacc gcattagatc ctttaagacc tgtaggagaa atcgcacaag ctaatggttt
     4501 atggttacat gtcgattcag ctatggcagg ttctgctatg atattaccag agtgtagatg
     4561 gatgtgggat ggtattgaat tggcagattc agtcgttgta aacgcacata agtggttggg
     4621 tgttgcattt gattgttcaa tatattacgt cagagatcca cagcacttaa ttagagtaat
     4681 gtcaaccaac ccatcttact tgcaatcagc agttgacggt gaagttaaga atttaaggga
     4741 ttggggtatc ccattaggta gaagatttag agctttgaaa ttatggttca tgttgaggtc
     4801 agagggtgtt gatgctttgc aagccagatt gagaagggat ttagataacg ctcaatggtt
     4861 agcaggtcaa gtagaagccg cagctgagtg ggaagttttg gcacctgttc aattgcaaac
     4921 cttgtgtatt aggcatagac cagcaggatt agagggtgag gctttagatg ctcatactaa
     4981 gggttgggct gaaagattaa acgcatcagg agcagcatac gtaacccctg ccacattaga
     5041 tggtagatgg atggtcaggg tttctatagg tgcattgcca actgaaagag gtgatgtaca
     5101 aagattgtgg gctagattac aagatgttat taagggagga tcctaactcg aggcgaattt
     5161 cttatgattt atgattttta ttattaaata agttataaaa aaaataagtg tatacaaatt
     5221 ttaaagtgac tcttaggttt taaaacgaaa attcttattc ttgagtaact ctttcctgta
     5281 ggtcaggttg ctttctcagg tatagcatga ggtcgctctt attgaccaca cctctaccgg
     5341 catgccgagc aaatgcctgc aaatcgctcc ccatttcgct gagcagttac agagatgtta
     5401 cgaaccacta gtgcactgca gtacacggtt tccttgaaat ttttttgatt cggtaatctc
     5461 cgaacagaag gaagaacgaa ggaaggagca cagacttaga ttggtatata tacgcatatg
     5521 tagtgttgaa gaaacatgaa attgcccagt attcttaacc caactgcaca gaacaaaaac
     5581 gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat aaggaacgtg ctgctactca
     5641 tcctagtcct gttgctgcca agctatttaa tatcatgcac gaaaagcaaa caaacttgtg
     5701 tgcttcattg gatgttcgta ccaccaagga attactggag ttagttgaag cattaggtcc
     5761 caaaatttgt ttactaaaaa cacatgtgga tatcttgact gatttttcca tggagggcac
     5821 agttaagccg ctaaaggcat tatccgccaa gtacaatttt ttactcttcg aagacagaaa
     5881 atttgctgac attggtaata cagtcaaatt gcagtactct gcgggtgtat acagaatagc
     5941 agaatgggca gacattacga atgcacacgg tgtggtgggc ccaggtattg ttagcggttt
     6001 gaagcaggcg gcagaagaag taacaaagga acctagaggc cttttgatgt tagcagaatt
     6061 gtcatgcaag ggctccctat ctactggaga atatactaag ggtactgttg acattgcgaa
     6121 gagcgacaaa gattttgtta tcggctttat tgctcaaaga gacatgggtg gaagagatga
     6181 aggttacgat tggttgatta tgacacccgg tgtgggttta gatgacaagg gagatgcatt
     6241 gggtcaacag tatagaaccg tggatgatgt ggtttctaca ggatctgaca ttattattgt
     6301 tggaagagga ctatttgcaa agggaaggga tgctaaggta gagggtgaac gttacagaaa
     6361 agcaggctgg gaagcatatt tgagaagatg cggccagcaa aactaaaaaa ctgtattata
     6421 agtaaatgca tgtatactaa actcacaaat tagagcttca atttaattat atcagttatt
     6481 acccgagtag agcacttgaa tccactgccc cgggaatctc ggtcgtaatg atttctataa
     6541 tgacgaaaaa aaaaaaattg gaaagaaaaa gcttcatggc ctttataaaa aggaactatc
     6601 caatacctcg ccagaaccaa gtaacagtat tttacggggc acaaatcaag aacaataaga
     6661 caggactgta aagatggacg cattgaactc caaagaacaa caagagttcc aaaaagtagt
     6721 ggaacaaaag caaatgaagg atttcatgcg tttgtactct aatctggtag aaagatgttt
     6781 cacagactgt gtcaatgact tcacaacatc aaagctaacc aataaggaac aaacatgcat
     6841 catgaagtgc tcagaaaagt tcttgaagca tagcgaacgt gtagggcagc gtttccaaga
     6901 acaaaacgct gccttgggac aaggcttggg ccgataaggt gtactggcgt atatatatct
     6961 aattatgtat ctctggtgta gcccattttt agcatgtaaa tataaagacc gagcggccgc
     7021 gtgttacaac caattaacca attctgatta gaaaaactca tcgagcatca aatgaaactg
     7081 caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga
     7141 aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat
     7201 tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc
     7261 aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa gcttatgcat
     7321 ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc
     7381 aaccaaaccg ttattcattc gtgattgcgc ctgagcgagg cgaaatacgc gatcgctgtt
     7441 aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc
     7501 aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg
     7561 gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg
     7621 aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc
     7681 aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg
     7741 atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc
     7801 agcatccatg ttggaattta atcgcggcct ggagcaagac gtttcccgtt gaatatggct
     7861 cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc atgatgatat
     7921 atttttatct tgtgcaatgt aacatcagag attttgagac acaacgtggc tttgttgaat
     7981 aaatcgaact tttgctgagt tgaaggatca gtcatgacca aaatccctta acgtgagttt
     8041 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     8101 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     8161 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     8221 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     8281 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     8341 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     8401 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     8461 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     8521 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     8581 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     8641 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     8701 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     8761 tctgtggata accgtgcggc cgc
//
