LOCUS       40924_43618        4611 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pTMH406, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4611)
  AUTHORS   Hsu TM, Welner DH, Russ ZN, Cervantes B, Prathuri RL, Adams PD, 
            Dueber JE.
  TITLE     Employing a biochemical protecting group for a sustainable indigo 
            dyeing strategy
  JOURNAL   Nat. Chem. Biol. (2018) In press
  PUBMED    29309053
REFERENCE   2  (bases 1 to 4611)
  AUTHORS   Hsu TM, Welner DH, Russ ZN, Cervantes B, Prathuri RL, Adams PD, 
            Dueber JE.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-AUG-2017) Bioengineering, UC Berkeley, 2151 Berkeley 
            Way Room 512D, Berkeley, CA 94704, USA
REFERENCE   3  (bases 1 to 4611)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4611)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. Chem. 
            Biol. (2018) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (20-AUG-2017) Bioengineering, UC Berkeley, 2151 Berkeley Way Room 
            512D, Berkeley, CA 94704, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4611
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(52..73)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(89..1168)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(1169..1246)
                     /label=lacI promoter
     promoter        1555..1573
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1574..1598
                     /label=lac operator
                     /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)."
     RBS             1613..1635
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1642..3018
                     /codon_start=1
                     /product="flavin-containing monooxygenase FMO"
                     /EC_number="1.14.13.8"
                     /label=flavin-containing monooxygenase FMO
                     /note="from Methylophaga aminisulfidivorans"
                     /protein_id="AUO17141.1"
                     /translation="MATRIAILGAGPSGMAQLRAFQSAQEKGAEIPELVCFEKQADWGG
                     QWNYTWRTGLDENGEPVHSSMYRYLWSNGPKECLEFADYTFDEHFGKPIASYPPREVLW
                     DYIKGRVEKAGVRKYIRFNTAVRHVEFNEDSQTFTVTVQDHTTDTIYSEEFDYVVCCTG
                     HFSTPYVPEFEGFEKFGGRILHAHDFRDALEFKDKTVLLVGSSYSAEDIGSQCYKYGAK
                     KLISCYRTAPMGYKWPENWDERPNLVRVDTENAYFADGSSEKVDAIILCTGYIHHFPFL
                     NDDLRLVTNNRLWPLNLYKGVVWEDNPKFFYIGMQDQWYSFNMFDAQAWYARDVIMGRL
                     PLPSKEEMKADSMAWREKELTLVTAEEMYTYQGDYIQNLIDMTDYPSFDIPATNKTFLE
                     WKHHKKENIMTFRDHSYRSLMTGTMAPKHHTPWIDALDDSLEAYLSDKSEIPVAKEAGS
                     "
     terminator      3087..3134
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     regulatory      complement(3190..3216)
                     /regulatory_class="terminator"
     CDS             complement(3220..4032)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     regulatory      complement(4033..4180)
                     /regulatory_class="promoter"
     rep_origin      complement(4197..4580)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 actggccgat aattgcagac gaacgggcat cggtcgagat cccggtgcct aatgagtgag
       61 ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
      121 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca
      181 gggtggtttt tcttttcacc agtgaaacgg gcaacagctg attgcccttc accgcctggc
      241 cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt
      301 tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta
      361 ccgagatatc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg
      421 ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca
      481 tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa
      541 tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac
      601 ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc
      661 ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga
      721 catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt
      781 catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg
      841 ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca
      901 gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac
      961 tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt
     1021 tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa
     1081 cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg
     1141 cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc
     1201 gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc
     1261 tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc
     1321 accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg
     1381 gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga
     1441 tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg
     1501 atgccggcca cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg aaattaatac
     1561 gactcactat aggggaattg tgagcggata acaattcccc tctagaaata attttgttta
     1621 actttaagaa ggagaagatc tatggccact aggattgcta tcttaggtgc tggtccttct
     1681 ggaatggctc aattgagagc atttcagtct gcccaggaga agggtgccga aattcctgag
     1741 ttagtctgct tcgaaaagca ggccgattgg ggtggtcaat ggaattacac atggaggacc
     1801 ggattggacg aaaatggtga acctgtacat tcatctatgt atagatattt gtggtctaac
     1861 ggacctaaag agtgcttgga gtttgccgat tatacctttg atgaacattt tggaaaacca
     1921 atcgcatctt acccaccaag agaagtatta tgggactaca ttaagggtag agttgagaaa
     1981 gctggtgtta ggaaatatat cagatttaat accgccgtta gacatgtcga atttaacgag
     2041 gactcacaaa ccttcaccgt tactgtccag gaccatacca ctgacactat atattcagaa
     2101 gagtttgatt acgttgtttg ttgcactggt catttctcta ccccatacgt tccagaattt
     2161 gagggatttg agaagttcgg tggaagaata ttgcatgccc acgattttag agatgcctta
     2221 gagtttaaag acaagaccgt tttattagta ggatcttcat actcagctga agacatcggt
     2281 tctcaatgtt acaaatacgg tgccaagaag ttaatttctt gttatagaac agcaccaatg
     2341 ggatataaat ggcctgagaa ctgggatgaa agaccaaatt tggttagggt cgacacagaa
     2401 aatgcctact ttgctgatgg ttcttctgag aaagtagatg ctattatatt atgtacaggt
     2461 tacattcatc acttcccatt cttaaatgat gacttgaggt tagttactaa caacaggtta
     2521 tggccattaa atttatataa gggagtagta tgggaggata accctaagtt cttctacatt
     2581 ggaatgcagg atcagtggta ttcattcaat atgtttgacg cacaggcttg gtatgctagg
     2641 gatgtaatca tgggtagatt acctttgcca tctaaggagg aaatgaaagc tgattcaatg
     2701 gcttggagag aaaaggagtt aacattggtt accgctgagg agatgtatac ataccagggt
     2761 gactatattc agaatttgat cgacatgact gattatccat ctttcgatat ccctgccacc
     2821 aataaaacct ttcttgagtg gaaacatcat aagaaagaaa acattatgac attcagggac
     2881 cattcatata ggtcattaat gacaggtact atggcaccta aacaccatac tccatggatt
     2941 gacgccttgg atgattcatt agaagcttac ttatctgaca agtcagaaat ccctgttgcc
     3001 aaggaggctg gatcctaatg atcaatccgg ctgctaacaa agcccgaaag gaagctgagt
     3061 tggctgctgc caccgctgag caataactag cataacccct tggggcctct aaacgggtct
     3121 tgaggggttt tttgctgagc agttacagag atgttacgaa ccactagtgc actgcagtac
     3181 agcggccgcg tgttacaacc aattaaccaa ttctgattag aaaaactcat cgagcatcaa
     3241 atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
     3301 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg
     3361 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
     3421 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
     3481 cttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
     3541 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgaggc gaaatacgcg
     3601 atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
     3661 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
     3721 tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
     3781 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac
     3841 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
     3901 atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
     3961 atataaatca gcatccatgt tggaatttaa tcgcggcctg gagcaagacg tttcccgttg
     4021 aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca
     4081 tgatgatata tttttatctt gtgcaatgta acatcagaga ttttgagaca caacgtggct
     4141 ttgttgaata aatcgaactt ttgctgagtt gaaggatcag ccatggctaa ttcccatgtc
     4201 agccgttaag tgttcctgtg tcactcaaaa ttgctttgag aggctctaag ggcttctcag
     4261 tgcgttacat ccctggcttg ttgtccacaa ccgttaaacc ttaaaagctt taaaagcctt
     4321 atatattctt ttttttctta taaaacttaa aaccttagag gctatttaag ttgctgattt
     4381 atattaattt tattgttcaa acatgagagc ttagtacgtg aaacatgaga gcttagtacg
     4441 ttagccatga gagcttagta cgttagccat gagggtttag ttcgttaaac atgagagctt
     4501 agtacgttaa acatgagagc ttagtacgtg aaacatgaga gcttagtacg tactatcaac
     4561 aggttgaact gcggatcttg gcggccgccc ctgaattcgc atctagactg a
//
