LOCUS       40924_43623        6080 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pTMH409, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6080)
  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 6080)
  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 6080)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6080)
  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..6080
                     /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="AUO17145.1"
                     /translation="MATRIAILGAGPSGMAQLRAFQSAQEKGAEIPELVCFEKQADWGG
                     QWNYTWRTGLDENGEPVHSSMYRYLWSNGPKECLEFADYTFDEHFGKPIASYPPREVLW
                     DYIKGRVEKAGVRKYIRFNTAVRHVEFNEDSQTFTVTVQDHTTDTIYSEEFDYVVCCTG
                     HFSTPYVPEFEGFEKFGGRILHAHDFRDALEFKDKTVLLVGSSYSAEDIGSQCYKYGAK
                     KLISCYRTAPMGYKWPENWDERPNLVRVDTENAYFADGSSEKVDAIILCTGYIHHFPFL
                     NDDLRLVTNNRLWPLNLYKGVVWEDNPKFFYIGMQDQWYSFNMFDAQAWYARDVIMGRL
                     PLPSKEEMKADSMAWREKELTLVTAEEMYTYQGDYIQNLIDMTDYPSFDIPATNKTFLE
                     WKHHKKENIMTFRDHSYRSLMTGTMAPKHHTPWIDALDDSLEAYLSDKSEIPVAKEAGS
                     "
     regulatory      3027..3038
                     /regulatory_class="ribosome_binding_site"
     RBS             3027..3038
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3045..4478
                     /codon_start=1
                     /product="PtUGT72B29 indoxyl glucosyltransferase"
                     /label=PtUGT72B29 indoxyl glucosyltransferase
                     /note="from Polygonum tinctorium"
                     /protein_id="AUO17143.1"
                     /translation="MESPAAPPTTAPPPHVIIVPSAGMGHLIPLAEFAKRLLPRFTFTF
                     AVPTSGPPSSSQRDFLSSLPASIDTSFLPEVDLSDAPSDAQIETLMSLMVVRSLPSLRD
                     LIASYSASGRRVAALVVDLFATDAIDVALELGIRPFIFFPSTAMTLSFFLHLEKLDETV
                     SCEFAELSDPVQIPGCIPVHGKDLIDPVQDRKNDAYKWLLHHSKRYKLAEGVIVNSFEG
                     LEGGPIRELLHPEPGKPRVYPVGPLIQAGSCEKGAAARPECLKWLDQQPRGSVLFVNFG
                     SGGVLSTEQQNELAGVLAHSQQRFLWVVRPPNDGIANATYFSVDGEIDPLKLLPEGFLE
                     QTAGRGLVLPMWAPQIDVLSHESTGGFLTHCGWNSTLESVFHGVPLITWPLYAEQKMNA
                     VMLTEGLRVGLRPSVGKDGIIRGAEIARVIGELMEGEEGKRIRSKMQELKRAASAVLSK
                     DGSSTRALEEVAKIWESKV"
     terminator      4556..4603
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     regulatory      complement(4659..4685)
                     /regulatory_class="terminator"
     CDS             complement(4689..5501)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     regulatory      complement(5502..5649)
                     /regulatory_class="promoter"
     rep_origin      complement(5666..6049)
                     /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 gatcctaacc aacgacaaag aggagaaaag atctatggaa tcccccgccg
     3061 ctccaccaac caccgctcca ccaccgcacg tcataatcgt gccctccgcc ggcatgggcc
     3121 acctcatccc cctcgccgag ttcgccaagc gcctccttcc gcgcttcacc ttcaccttcg
     3181 ccgtacccac cagcggcccg ccctcatcct cccagcgcga cttcctctcc tccctccctg
     3241 cctccatcga cacctccttc ctccccgagg tcgacctctc cgacgccccc tccgacgccc
     3301 aaatcgagac tctcatgtcc ctcatggttg tccgctccct cccctcgctc cgcgacctca
     3361 ttgcctccta ctccgcctcc ggccgccgcg tcgccgccct cgtcgtcgac cttttcgcca
     3421 ctgatgcaat cgacgtcgcc cttgagctcg gcatccgccc tttcatcttc ttcccctcca
     3481 ccgccatgac cctctccttc ttcctccacc tcgagaagct tgatgaaacg gtgtcatgtg
     3541 agtttgccga gctgtccgac ccggttcaga tccccgggtg tattccggtc cacggcaagg
     3601 atttgatcga cccggttcag gataggaaga acgacgccta caagtggctc ctccaccact
     3661 ccaagaggta taaattggcc gagggtgtta tcgtaaatag cttcgagggt ttggaggggg
     3721 gaccgatcag ggagcttttg caccccgagc cgggaaagcc gcgggtttac ccggtcggac
     3781 cgctgattca ggccggttcg tgcgagaagg gggcagctgc ccggcctgag tgcttgaagt
     3841 ggttggacca gcagccacgt ggatccgtcc tattcgtgaa tttcgggagt ggtggggtcc
     3901 tcagtacgga gcagcagaac gagcttgcag gtgtgctggc ccacagccag cagaggttcc
     3961 tatgggtggt taggcctcca aacgacggca ttgccaacgc cacgtatttc agcgtcgacg
     4021 gggagatcga cccgttgaaa ctcctgcccg aggggttctt ggagcagacc gcgggcaggg
     4081 gtttggtctt gccaatgtgg gccccgcaga tcgatgtctt aagccatgag tcgacgggcg
     4141 ggttcttgac gcattgtggg tggaattcaa cactggagag cgtgttccat ggggtaccac
     4201 taattacatg gcccctctat gcagagcaaa agatgaacgc tgttatgcta accgagggcc
     4261 tgagggtggg actcagaccc tcagtgggta aggatggaat catccgaggt gctgagatcg
     4321 cacgagttat aggggagttg atggaaggtg aggaagggaa acgaatacgg agtaagatgc
     4381 aggagttgaa gcgtgcggct tctgctgtat tgagcaaaga tggatcatct actcgagctc
     4441 ttgaagaggt tgcaaaaatt tgggaaagca aggtttaagg atcctaatga tcaatccggc
     4501 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
     4561 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgagca gttacagaga
     4621 tgttacgaac cactagtgca ctgcagtaca gcggccgcgt gttacaacca attaaccaat
     4681 tctgattaga aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta
     4741 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
     4801 ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
     4861 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
     4921 acgactgaat ccggtgagaa tggcaaaagc ttatgcattt ctttccagac ttgttcaaca
     4981 ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt
     5041 gattgcgcct gagcgaggcg aaatacgcga tcgctgttaa aaggacaatt acaaacagga
     5101 atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca
     5161 ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat
     5221 gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc
     5281 cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
     5341 agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc
     5401 ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat
     5461 cgcggcctgg agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg
     5521 tttatgtaag cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtaa
     5581 catcagagat tttgagacac aacgtggctt tgttgaataa atcgaacttt tgctgagttg
     5641 aaggatcagc catggctaat tcccatgtca gccgttaagt gttcctgtgt cactcaaaat
     5701 tgctttgaga ggctctaagg gcttctcagt gcgttacatc cctggcttgt tgtccacaac
     5761 cgttaaacct taaaagcttt aaaagcctta tatattcttt tttttcttat aaaacttaaa
     5821 accttagagg ctatttaagt tgctgattta tattaatttt attgttcaaa catgagagct
     5881 tagtacgtga aacatgagag cttagtacgt tagccatgag agcttagtac gttagccatg
     5941 agggtttagt tcgttaaaca tgagagctta gtacgttaaa catgagagct tagtacgtga
     6001 aacatgagag cttagtacgt actatcaaca ggttgaactg cggatcttgg cggccgcccc
     6061 tgaattcgca tctagactga
//
