LOCUS       V008056                 7721 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V008056
VERSION     V008056
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7721)
  AUTHORS   Quandt EM, Hammerling MJ, Summers RM, Otoupal PB, Slater B, Alnahhas
            RN, Dasgupta A, Bachman JL, Subramanian MV, Barrick JE.
  TITLE     Decaffeination and measurement of caffeine content by addicted E.
            coli with a refactored N-demethylation operon
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7721)
  AUTHORS   Quandt EM, Hammerling MJ, Summers RM, Otoupal PB, Slater B, Alnahhas
            RN, Dasgupta A, Bachman JL, Subramanian MV, Barrick JE.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-FEB-2013) Chemistry and Biochemistry, The University
            of Texas at Austin, 2500 Speedway A4800, Austin, TX 78712-1639, USA
REFERENCE   3  (bases 1 to 7721)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7721)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (14-FEB-2013) Chemistry and Biochemistry, The University of Texas at
            Austin, 2500 Speedway A4800, Austin, TX 78712-1639, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7721
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      22..56
                     /gene="ndmA"
                     /note="BBa_J23100; derived from BBa_K515105"
                     /regulatory_class="promoter"
     regulatory      65..89
                     /label="derived from BBa_K515105"
                     /note="derived from BBa_K515105"
                     /regulatory_class="ribosome_binding_site"
     RBS             74..82
                     /label="Shine-Dalgarno sequence"
                     /note="full consensus sequence for ribosome-binding sites
                     upstream of start codons in E. coli; complementary to a
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             90..1142
                     /gene="ndmA"
                     /label="Methylxanthine N1-demethylase NdmA"
                     /note="Methylxanthine N1-demethylase NdmA from Pseudomonas
                     putida. Accession#: H9N289"
     regulatory      1153..1164
                     /label="BBa_B0034"
                     /note="BBa_B0034"
                     /regulatory_class="ribosome_binding_site"
     RBS             1153..1164
                     /note="strong bacterial ribosome binding site (Elowitz and
                     Leibler, 2000)"
     CDS             1171..2235
                     /gene="ndmB"
                     /label="Methylxanthine N3-demethylase NdmB"
                     /note="Methylxanthine N3-demethylase NdmB from Pseudomonas
                     putida. Accession#: H9N290"
     regulatory      2256..2267
                     /label="BBa_B0034"
                     /note="BBa_B0034"
                     /regulatory_class="ribosome_binding_site"
     RBS             2256..2267
                     /note="strong bacterial ribosome binding site (Elowitz and
                     Leibler, 2000)"
     CDS             2274..3128
                     /codon_start=1
                     /gene="ndmC"
                     /product="ndmC"
                     /label="ndmC"
                     /protein_id="AGL91158.1"
                     /translation="MSTDQVIFNDWHPVAALEDVSLDKRYRCRLLGRTVSYVKTSDAVN
                     AHWEESADEIKTIRAKEIYGLLWLSFADKPSEMFDIAEFKEPDRRIVSAGSVRVNVSGL
                     RAIENFLDMAHFPFVHTDILGAEPLTEVEPYNVNYDETVDEIFATECKFPQPKGSATAV
                     EPIDMQYIYRITRPYSAILYKTCPPEPHRWDALGLFIQPVDEDWCIAHTIMCYVDDVNS
                     DQQLRHFQQTIFGQDLMILINQVPKRLPLAASRESPVRADVLATAYRRWLREKGVQYGA
                     LRD"
     gene            2274..3128
                     /gene="ndmC"
                     /label="ndmC"
     regulatory      3136..3147
                     /label="BBa_B0034"
                     /note="BBa_B0034"
                     /regulatory_class="ribosome_binding_site"
     RBS             3136..3147
                     /note="strong bacterial ribosome binding site (Elowitz and
                     Leibler, 2000)"
     CDS             3154..4917
                     /gene="ndmD"
                     /label="Oxidoreductase NdmD"
                     /note="Oxidoreductase NdmD from Pseudomonas putida.
                     Accession#: H9N291"
     regulatory      4965..4976
                     /label="BBa_B0034"
                     /note="BBa_B0034"
                     /regulatory_class="ribosome_binding_site"
     RBS             4965..4976
                     /note="strong bacterial ribosome binding site (Elowitz and
                     Leibler, 2000)"
     CDS             4983..5657
                     /codon_start=1
                     /gene="gst9"
                     /product="gst9"
                     /label="gst9"
                     /protein_id="AGL91156.1"
                     /translation="MITLYDYELSGNCYKVRMLLSILKLPYTIETVEFFPSREHKSPAF
                     LRINPLGQLPVLRDDDFVLRDAQAILVYLANKYDTSLTWYPTHAPAILGQANMWLAFAD
                     GLTGSISAARLHDLFFYDFNAEQCRARAHELLRILDEHLWAAEQNGHQWLCPAAAPTIA
                     DLACFPYIALSDEAGVSLLDYPAVRRWLDRVKRIPGFTVMSGIFPTSASFEPASLAEET
                     CQ"
     gene            4983..5657
                     /gene="gst9"
                     /label="gst9"
     misc_feature    5673..5693
                     /label="BioBrick suffix"
                     /note="universal suffix for all parts"
     terminator      5694..5751
                     /label="his operon terminator"
                     /note="This putative transcriptin terminator from the E.
                     coli his operon has a 2-bp deletion introduced during
                     synthesis. Its efficiency has not been determined."
     rep_origin      complement(5946..6534)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     terminator      complement(6717..6811)
                     /label="lambda t0 terminator"
                     /note="transcription terminator from phage lambda"
     CDS             complement(6835..7491)
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(7492..7595)
                     /label="cat promoter"
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     terminator      complement(7675..7718)
                     /label="bacterial terminator"
                     /note="putative bacterial transcription terminator"
     misc_feature    7721
                     /label="BioBrick prefix"
                     /note="BioBrick prefix for parts that do not start with
                     'ATG'"
ORIGIN
        1 aattcgcggc cgcttctaga gttgacggct agctcagtcc taggtacagt gctagctact
       61 agagaaatca aattaaggag gtaagataaa tggaacaggc aatcattaat gatgaacggg
      121 agtatcttcg ccatttctgg catcccgtat gtactgtaac tgagcttgaa aaggcgcatc
      181 cttccagcct cggccccctg gccgttaagc tgctgaatga acagctcgtt gtcgccaagc
      241 taggcgatga gtacgtcgcg atgcgtgata gatgcgctca tcgatcagct aagctttcct
      301 tgggtacagt aagtggaaac cgcctacagt gcccctatca cggatggcaa tatgatacgc
      361 atggcgcttg ccagctcgta ccagcgtgcc ccaacagccc aatacccaac aaggctaaag
      421 ttgatcgctt cgattgcgaa gaacgctatg gattgatttg gattcgattg gactctagtt
      481 ttgactgcac tgaaattccc tactttagtg cagccaacga tcctaggttg cgtattgtta
      541 tacaagaacc ttactggtgg gatgcgactg cagaacgtag atgggaaaat tttacagatt
      601 tttctcactt tgcattcatt cacccaggca cgcttttcga tccaaataat gctgaacctc
      661 caattgttcc gatggatcga ttcaatggtc agtttcggtt tgtctacgac acgccagaag
      721 atatggctgt cccaaatcag gctccaattg gttcattttc gtatacttgc agcatgccgt
      781 ttgctattaa ccttgaagta tccaaatact ccagcagttc gctgcatgtg ttattcaatg
      841 tgtcatgtcc ggtagacagc cacaccacga aaaactttct gatcttcgct agggagcaat
      901 cggacgactc ggattatctg cacattgcat ttaatgatct cgtcttcgct gaagacaaac
      961 cagtaattga gtcccaatgg cctaaagacg cgccagcaga tgaggtctca gtagtcgcag
     1021 ataaggtatc gatacaatat agaaaatggc tgcgggaact aaaagaagct cataaagaag
     1081 gttcacaagc cttccgaagt gctttgttag acccagtcat tgaaagcgat aggagctaca
     1141 tataatctag agaaagagga gaaatactag atgaaagaac agctcaagcc gctgctagaa
     1201 gacaagactt accttcgcca cttctggcat cccgtgtgta cccttaatga attcgaacgc
     1261 gccaacgcca gtgggcacgg ccccatgggc gtcaccttgc taggcgagaa attggtgttg
     1321 gccaggttaa attcaaagat cattgcggct gctgaccgat gtgctcatcg atcggcacag
     1381 ctctccatcg gccgcgtttg cagcaacgcc ggaaaggact atctcgaatg cccgtatcac
     1441 ggctggcgct acgatgaggc tggggcctgt caactgatcc ctgcttgccc tgacaaaagc
     1501 atctctcctc gggccaagat ttcctcattc gattgtgagg tgaaatacga catcgtgtgg
     1561 gtacggctgg ataacagctt cgattgcact cagattccat acctcagcga tttcgataat
     1621 cccgacatgc aggtaatcgt tgccgattcg tatatttggg agactgttgc cgagcggcgg
     1681 tgggagaact ttacagattt ttcgcacttt gccttcgtac acccagggac gctctatgat
     1741 ccgtttttcg ctagccaccc aactgtttac gtgaatcgcg ttgatggtga gttgcaattc
     1801 aaactcgctc cgccgcgtga aatgaaaggc atcccgccag aagcaccgat gggtgacttc
     1861 acctaccgct gcacaatgcc gtattcagta aatcttgaaa tcaaattgtg gaaagatgac
     1921 tcacgtttcg ttctttggac taccgctagc cctgtcgata acaagtcttg ccggaatttt
     1981 atgattattg tgcgtgagaa ggataatcaa cctgatcata tgcacctggc tttccagaag
     2041 cgggtgcttg acgaagacca gcctgttatc gaatcgcaat ggcctctcga aatacagacc
     2101 tcggaagtct ccgttgcaac cgataaaatt tccgtccagt tccgcaaatg gcataaagag
     2161 ctatctctgt cagccgttga aggacgggag gcgttccgcg attccgtctt gacgaatgtt
     2221 attgaagaag aacagtaagc gagaccggtc tagagaaaga ggagaaatac tagatgtcta
     2281 ctgaccaagt aatttttaac gactggcatc cagtcgctgc tttggaagat gtatctctcg
     2341 ataaacgata ccgctgtcga ctactaggtc gtacagttag ttatgtgaaa acatctgatg
     2401 ctgttaatgc tcattgggaa gaaagtgcag atgaaattaa gaccatccgc gctaaagaaa
     2461 tctatggtct tctgtggctc tcctttgccg acaaacccag tgagatgttc gatattgcag
     2521 agttcaagga acctgatcgc cgaatcgtca gcgctggatc tgtgcgcgta aatgtctcag
     2581 gactgcgtgc tatcgaaaac tttctggaca tggctcattt cccttttgtt catacggata
     2641 ttttgggtgc agagccactg acggaagttg agccgtataa cgtcaactat gatgaaactg
     2701 ttgatgagat cttcgctact gagtgtaagt tcccgcaacc taaaggctca gctactgctg
     2761 tcgagccaat tgatatgcag tatatatacc gcattacacg gccatattcc gccattttat
     2821 ataaaacttg cccgcccgaa ccgcatagat gggatgccct cggtctgttt attcaacctg
     2881 tagacgaaga ttggtgcata gcgcatacaa ttatgtgcta tgtggatgac gttaactcag
     2941 atcaacaact tcgccatttc cagcagacga tttttggcca ggacttgatg attcttatca
     3001 accaagttcc aaagcgtctt cccctggctg caagtcgaga gagcccagtc cgggccgatg
     3061 tgcttgcaac agcttatcgt cgctggctgc gtgagaaagg tgtgcaatat ggtgctctgc
     3121 gggactaatc tagagaaaga ggagaaatac tagatgaaca aacttgacgt caaccagtgg
     3181 tttcctattg ctaccactga agatctcccg aagcgccatg tctttcatgc cacgttgttg
     3241 gggcaagaaa tggccatctg gcgcgatgac tctggttcag ttaatgcttg ggagaaccgc
     3301 tgcccgcata gaggattgcg gttgacactg ggtgctaata ccggtaacga gttgcggtgt
     3361 cagtatcatg gatggactta tgaaagcggg actggtggct gcacttttgt cccagcccat
     3421 cgcgatgcac cacccccaaa tgccgcgcgg gttaatactt ttcctgtccg cgaaaagcac
     3481 ggctttatct ggacgacatt aggtcagccg ccaggagagc ccatttcaat cctcgatgac
     3541 gctcagcttg taaacgctgt aaaaacaaat ctgcatagcg tagttataga tgctgatatt
     3601 gacggagttg tcagcgtcct acgtcagaat ctttcagcgt tcatcgatgt gtttggtgcg
     3661 gccagcgctg aagatctgca tttgaaatcc atgctgcaag atcgagggat tctggtaaca
     3721 agatcaggct ctattgctat tcatttttat atgcagcgct caaccattag taaatgcgtt
     3781 gtacatgcgc aagtacttac tccgggacgt ccaggatacg aacttcaaaa gaactactcg
     3841 tatgccatga acgttatccg cagggcagca gaagctgtag ctaccgactt gattagcatt
     3901 acagatatca gcgatcagac tatcgaaaag cttgaagtcg ttagagaaaa catgactaag
     3961 gctcctccaa cccactatat ctgcgaagtg gttacgcgta ctcaagagac aggtgatatt
     4021 aactcatact ggctgaagcc tatcggctac ccactaccag cattcagtcc agggatgcac
     4081 atcagcatca caacgccgga gggtagcatt cgacaatatt ccctcgtgaa cgggcctgac
     4141 gagcgtgaat ccttcatcat cggtgtgaag aaagagattc agtcccgtgg cggctccaga
     4201 tcaatgcacg aagatgtgaa ggttggaacg caactaaaag ttacacttcc gaggaacggt
     4261 tttccactcg tccaaaccag aaaacacccg attctcgtag caggtggcat cggtatcacc
     4321 ccaattttgt gtatggcaca ggctctggat cagcaaggtt catcgtatga aatacattat
     4381 tttgctcgtg catttgagca tgttccattc caggatcgac tgactgcgtt gggcgatcgt
     4441 ttgaatgtgc atcttggcct cggcccagac gagactagag caaaacttgc cgacatcatg
     4501 gagattcata acgcccaaga cgtagatgtt tacacttgcg gcccgcaacc aatgatcgaa
     4561 actgtatctg ctgtcgctct tgctcatggc atcgctgaag agtccatccg atttgaattt
     4621 ttcagtaaaa agaacgatgt tcccgtttct gatgaagaat atgaggttga gctcaaaaaa
     4681 actggtcaaa tattcactgt ctcgcctggc tctacgttgt tgcaagcttg tttggacaac
     4741 gatgttcgta tcgaagcttc ttgtgagcag ggtgtatgcg ggacttgtat aactccagtc
     4801 gtatccggcg atctcgagca tcatgacact tacctttcta agaaagaaag ggaaagcggt
     4861 aagtggatca tgccgtgtgt ttcgcgctgc aagtccaaaa aaatcgttct cgatctgtga
     4921 ggccgggcat cacccacggt atggacgagc tgtataaact agagaaagag gagaaatact
     4981 agatgatcac gctttatgac tatgaactct cggggaattg ctacaaggtg cggatgctcc
     5041 tttccatcct gaagctaccg tacacgatag agacagtgga atttttcccg tccagggaac
     5101 ataaatcacc ggccttcctg cgcatcaatc cactcggcca gttgccggta ttgcgcgatg
     5161 atgatttcgt gttgcgcgat gcccaggcca tcctggttta cctggcgaac aaatacgaca
     5221 ccagtttgac ctggtatccg acacatgcgc ccgccatcct cggccaggca aatatgtggc
     5281 tcgcgtttgc cgatggtttg accggctcga tttccgctgc acgtttgcat gatttgtttt
     5341 tttacgattt caatgcagag caatgccgtg cccgcgcgca cgaactactg cgcatactgg
     5401 acgaacacct gtgggcggcg gaacaaaatg gacatcaatg gctatgtccg gccgccgcac
     5461 ccaccatcgc ggatcttgcc tgcttcccgt atatcgcatt gtcggatgaa gccggcgttt
     5521 cactactcga ctacccggcg gtacgacgct ggctggaccg tgtcaaacgc atccccggat
     5581 tcaccgtcat gtcggggatt tttccaacct cagcttcctt cgagccggcc tcacttgcgg
     5641 aagaaacctg tcaataagta atgatactag agtactagta gcggccgctg cagtccggca
     5701 aaaaagggca aggtgtcacc accctgccct ttttctttaa aaccgaaaag attacttcgc
     5761 gttatgcagg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5821 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5881 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5941 gcgtttttcc acaggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     6001 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     6061 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     6121 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     6181 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     6241 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     6301 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     6361 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     6421 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     6481 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     6541 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     6601 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     6661 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagctcgag gcttggattc
     6721 tcaccaataa aaaacgcccg gcggcaaccg agcgttctga acaaatccag atggagttct
     6781 gaggtcatta ctggatctat caacaggagt ccaagcgagc tcgatatcaa attacgcccc
     6841 gccctgccac tcatcgcagt actgttgtaa ttcattaagc attctgccga catggaagcc
     6901 atcacaaacg gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt
     6961 ataatatttg cccatggtga aaacgggggc gaagaagttg tccatattgg ccacgtttaa
     7021 atcaaaactg gtgaaactca cccagggatt ggctgagacg aaaaacatat tctcaataaa
     7081 ccctttaggg aaataggcca ggttttcacc gtaacacgcc acatcttgcg aatatatgtg
     7141 tagaaactgc cggaaatcgt cgtggtattc actccagagc gatgaaaacg tttcagtttg
     7201 ctcatggaaa acggtgtaac aagggtgaac actatcccat atcaccagct caccgtcttt
     7261 cattgccata cgaaattccg gatgagcatt catcaggcgg gcaagaatgt gaataaaggc
     7321 cggataaaac ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa tatccagctg
     7381 aacggtctgg ttataggtac attgagcaac tgactgaaat gcctcaaaat gttctttacg
     7441 atgccattgg gatatatcaa cggtggtata tccagtgatt tttttctcca ttttagcttc
     7501 cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt
     7561 atggtgaaag ttggaacctc ttacgtgccc gatcaactcg agtgccacct gacgtctaag
     7621 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg cagaatttca
     7681 gataaaaaaa atccttagct ttcgctaagg atgatttctg g
//
