LOCUS       V003255                 7800 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003255
VERSION     V003255
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7800)
  AUTHORS   Hon S, Olson DG, Holwerda EK, Lanahan AA, Murphy SJL., Maloney MI,
            Zheng T, Papanek B, Guss AM, Lynd LR.
  TITLE     The ethanol pathway from Thermoanaerobacterium saccharolyticum
            improves ethanol production in Clostridium thermocellum
  JOURNAL   Metab. Eng. 42, 175-184 (2017)
   PUBMED   28663138
REFERENCE   2  (bases 1 to 7800)
  AUTHORS   Hon S, Olson DG, Holwerda EK, Lanahan AA, Murphy SJL., Maloney MI,
            Zheng T, Papanek BA, Guss AM, Lynd LR.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-MAR-2017) Thayer School of Engineering, Dartmouth
            College, 14 Engineering Drive, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 7800)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7800)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "Metab. Eng.
            42, 175-184 (2017)"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (27-MAR-2017) Thayer School of Engineering, Dartmouth College, 14
            Engineering Drive, Hanover, NH 03755, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7800
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..92
                     /label="cat promoter"
                     /note="cat promoter"
                     /regulatory_class="promoter"
     CDS             93..740
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     CDS             complement(824..1681)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(1682..1773)
                     /label="AmpR promoter"
     rep_origin      2033..2578
                     /direction=RIGHT
                     /label="p15A ori"
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells
                     that contain a second plasmid with the ColE1 origin."
     CDS             3049..4050
                     /label="repB"
                     /note="RepB replication protein"
     regulatory      4099..4307
                     /label="Clo1313_2638 promoter"
                     /note="Clo1313_2638 promoter"
                     /regulatory_class="promoter"
     CDS             4308..5162
                     /codon_start=1
                     /gene="nfnA"
                     /product="NfnA"
                     /label="nfnA"
                     /note="NADH-dependent reduced ferredoxin:NADP+
                     oxidoreductase"
                     /protein_id="ASK86044.1"
                     /translation="MNEILEKKQLNPTVKMMVINAPLMAKKAKPGQFVIVRVDEKGERI
                     PLTIADYDRNKGTITIIFQEVGMSTKKLGTLNVGDRLHDFVGPLGKPVEFSKDTKRVLA
                     IGGGVGVAPLYPKVKMLNEMKVPVDSIIGGRSAEYVILEDEMKKVSENLYITTDDGTKG
                     RKGFVTDVLKELIEKDNKYDEVIAIGPLIMMKMVCNITKEYNIPTMVSMNPIMIDGTGM
                     CGGCRVTVGGETKFACVDGPAFDGLKVDFDEAMRRQNMYKDMERKVLENYEHECKLGGI
                     LNG"
     gene            4308..5162
                     /gene="nfnA"
                     /label="nfnA"
     CDS             5155..6546
                     /codon_start=1
                     /gene="nfnB"
                     /product="NfnB"
                     /label="nfnB"
                     /note="NADH-dependent reduced ferredoxin:NADP+
                     oxidoreductase"
                     /protein_id="ASK86045.1"
                     /translation="MANMSLKKVPMPEQEPDQRNKNFKEVALGYEENMAVEEAERCIQC
                     KNQPCVEGCPVHVKIPEFIKLIANRDFEGAYQKIKETNNLPAICGRVCPQESQCESVCT
                     RGKKGEPVAIGRLERFTADWHMKNNEDKIEKPETNGRKVAVIGSGPAGLSCAGDLAKMG
                     YDTTIFEAFHTPGGVLMYGIPEFRLPKEIVQKEIDSLKKLGVKIETNMVIGKILTIDDL
                     FDMGYEAVFIGTGAGLPKFMNIPGENLNGVYSANEFLTRINLMKAYDFPNSPTPVKVGK
                     KVAVVGGGNVAMDAARSAKRMGAEEVYIVYRRSEEEMPARLEEIHHAKEEGIIFKLLTN
                     PVRIIGDESGSVKGIECVNMVLGDVDESGRRRPVEEKGSEHVIDVDTVIIAIGQSPNPL
                     ITSTTEGLEKQRWGGIIVNEETLETSRRGVFAGGDAVTGAATVILAMGAGKKAAASIHK
                     YLSEK"
     gene            5155..6546
                     /gene="nfnB"
                     /label="nfnB"
     CDS             6564..7763
                     /codon_start=1
                     /gene="adhA"
                     /product="AdhA"
                     /label="adhA"
                     /note="NADPH-dependent alcohol dehydrogenase"
                     /protein_id="ASK86043.1"
                     /translation="MWETKVNPSKIFELRCKNTTYFGVGSIHKIKDILENLKINGINNV
                     IFITGKGSYKTSGAWDVVRPVLEELDLKYSLYDKVGPNPTVDMIDEAAKIGRESGAKAV
                     IGIGGGSPIDTAKSVAVLLKYTDKNARELYKQKFIPDDAVPIIAINLTHGTGTEVDRFA
                     VATIPEKNYKPAIAYDCLYPMFAIDDPSLMTKLDKKQTIAVTVDALNHITEAATTLVAS
                     PYSILTAKETVRLIVRYLPAAVNDPLNIVARYYLLYASALAGISFDNGLLHLTHALEHP
                     LSAVKPEIAHGLGLGAILPAVIKAIYPATAEVLADVYSPIVPGLKGLPVEAEYVAEKVQ
                     EWLFSVGCIQKLSDFGFTKDDIPNLVKLAKTTPSLDGLLSIAPVEATESVIEKIYLKSL
                     "
     gene            6564..7763
                     /gene="adhA"
                     /label="adhA"
ORIGIN
        1 ttgactttta aaaaaggatt gattctaatg aagaaagcag acaagtaagc ctcctaaatt
       61 cactttagat aaaaatttag gaggcatatc aaatgaactt taataaaatt gatttagaca
      121 attggaagag aaaagagata tttaatcatt atttgaacca acaaacgact tttagtataa
      181 ccacagaaat tgatattagt gttttatacc gaaacataaa acaagaagga tataaatttt
      241 accctgcatt tattttctta gtgacaaggg tgataaactc aaatacagct tttagaactg
      301 gttacaatag cgacggagag ttaggttatt gggataagtt agagccactt tatacaattt
      361 ttgatggtgt atctaaaaca ttctctggta tttggactcc tgtaaagaat gacttcaaag
      421 agttttatga tttatacctt tctgatgtag agaaatataa tggttcgggg aaattgtttc
      481 ccaaaacacc tatacctgaa aatgcttttt ctctttctat tattccatgg acttcattta
      541 ctgggtttaa cttaaatatc aataataata gtaattacct tctacccatt attacagcag
      601 gaaaattcat taataaaggt aattcaatat atttaccgct atctttacag gtacatcatt
      661 ctgtttgtga tggttatcat gcaggattgt ttatgaactc tattcaggaa ttgtcagata
      721 ggcctaatga ctggctttta taatatgaga taatgccgac tgtacttttt acagtcggtt
      781 ttctaatgtc actagggctc gcctttggga agtttgaagg ttaccaatgc ttaatcagtg
      841 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
      901 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
      961 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1021 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1081 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctgcag
     1141 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1201 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1261 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1321 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1381 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac
     1441 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1501 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1561 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1621 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1681 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1741 acatatttga atgtatttag aaaaataaac aaaagagttt gaggaaaact taagaaaacg
     1801 ggaaactatt tcctgttgag aatgtgaaat taaatgataa aaagtgatat aatagggtca
     1861 ggcgtacaga agcctgacct ttttgcaaaa ttgttttatt ggcgcggcgt atatatacag
     1921 catatataca atgtaaattt atacaatcaa attgttgata caatcgaatt atgcggctaa
     1981 aaatttgctc atgagcctag aaatatttta tctgattaat aagatgatct tcttgagatc
     2041 gttttggtct gcgcgtaatc tcttgctctg aaaacgaaaa aaccgccttg cagggcggtt
     2101 tttcgaaggt tctctgagct accaactctt tgaaccgagg taactggctt ggaggagcgc
     2161 agtcaccaaa acttgtcctt tcagtttagc cttaaccggc gcatgacttc aagactaact
     2221 cctctaaatc aattaccagt ggctgctgcc agtggtgctt ttgcatgtct ttccgggttg
     2281 gactcaagac gatagttacc ggataaggcg cagcggtcgg actgaacggg gggttcgtgc
     2341 atacagtcca gcttggagcg aactgcctac ccggaactga gtgtcaggcg tggaatgaga
     2401 caaacgcggc cataacagcg gaatgacacc ggtaaaccga aaggcaggaa caggagagcg
     2461 cacgagggag ccgccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2521 ccactgattt gagcgtcaga tttcgtgatg cttgtcaggg gggcggagcc tatggaaaaa
     2581 cggctttgcc gcggccctct cacttccctg ttaagtatct tcctggcatc ttccaggaaa
     2641 tctccgcccc gttcgtaagc catttccgct cgccgcagtc gaacgaccga gcgtagcgag
     2701 tcagtgagcg aggaagcgga atatatcctg tatcacatat tctgctgacg caccggtgca
     2761 gccttttttc tcctgccaca tgaagcactt cactgacacc ctcatcagtg ccaacatagt
     2821 aagccagtat acaaatgagt gctttttttg cgttttgagc gtagcgaaaa acgagttctt
     2881 tctattcttg atacatatag aaataacgtc atttttattt tagttgctga aaggtgcgtt
     2941 gaagtgttgg tatgtatgtg ttttaaagta ttgaaaaccc ttaaaattgg ttgcacagaa
     3001 aaaccccatc tgttaaagtt ataagtgacc aaacaaataa ctaaatagat gggggtttct
     3061 tttaatatta tgtgtcctaa tagtagcatt tattcagatg aaaaatcaag ggttttagtg
     3121 gacaagacaa aaagtggaaa agtgagacca tggagagaaa agaaaatcgc taatgttgat
     3181 tactttgaac ttctgcatat tcttgaattt aaaaaggctg aaagagtaaa agattgtgct
     3241 gaaatattag agtataaaca aaatcgtgaa acaggcgaaa gaaagttgta tcgagtgtgg
     3301 ttttgtaaat ccaggctttg tccaatgtgc aactggagga gagcaatgaa acatggcatt
     3361 cagtcacaaa aggttgttgc tgaagttatt aaacaaaagc caacagttcg ttggttgttt
     3421 ctcacattaa cagttaaaaa tgtttatgat ggcgaagaat taaataagag tttgtcagat
     3481 atggctcaag gatttcgccg aatgatgcaa tataaaaaaa ttaataaaaa tcttgttggt
     3541 tttatgcgtg caacggaagt gacaataaat aataaagata attcttataa tcagcacatg
     3601 catgtattgg tatgtgtgga accaacttat tttaagaata cagaaaacta cgtgaatcaa
     3661 aaacaatgga ttcaattttg gaaaaaggca atgaaattag actatgatcc aaatgtaaaa
     3721 gttcaaatga ttcgaccgaa aaataaatat aaatcggata tacaatcggc aattgacgaa
     3781 actgcaaaat atcctgtaaa ggatacggat tttatgaccg atgatgaaga aaagaatttg
     3841 aaacgtttgt ctgatttgga ggaaggttta caccgtaaaa ggttaatctc ctatggtggt
     3901 ttgttaaaag aaatacataa aaaattaaac cttgatgaca cagaagaagg cgatttgatt
     3961 catacagatg atgacgaaaa agccgatgaa gatggatttt ctattattgc aatgtggaat
     4021 tgggaacgga aaaattattt tattaaagag tagttcaacc cggggatcct ctagagtcga
     4081 cctgcaggca tgcaagctga taaacaaagg acggttcagg gcttctgctc atcctactct
     4141 gcattgtaaa aaggtaggat gaatttttat ttttaatctt attgaaaaaa atttttgaaa
     4201 atcggtttta ttaaaaaaaa gtgggtatat ttataatagt caattgattg gttaaaaaaa
     4261 tttaaataag caaacagaat aataacaaaa gtaaggagga atttgttatg aacgagattt
     4321 tagagaagaa gcaattgaat cccactgtca aaatgatggt tataaacgct ccactaatgg
     4381 cgaaaaaggc gaaacctgga cagtttgtca tagtaagagt agatgaaaag ggtgaacgca
     4441 ttcctttgac aatagcggat tacgacagaa acaaaggcac gattactata atatttcaag
     4501 aagtcggcat gagcaccaag aagttaggta cgttaaacgt aggcgataga ttgcatgact
     4561 tcgtaggacc tttaggaaaa cctgttgaat tttcaaaaga tacgaagaga gttttagcaa
     4621 tcggcggagg tgttggagtt gcgccgcttt atccaaaagt aaaaatgcta aatgagatga
     4681 aagtaccagt ggatagtata ataggcggca gaagtgcaga gtacgtcata ttagaagatg
     4741 aaatgaaaaa agtcagtgaa aacctttata taacgacgga tgacggcaca aaaggccgta
     4801 aaggctttgt aacagatgta ttaaaagaat tgatagaaaa agacaacaaa tacgatgaag
     4861 taattgcaat tgggccactg ataatgatga agatggtatg caatataaca aaagaataca
     4921 acataccaac tatggtcagc atgaacccga taatgataga cggaactgga atgtgcggag
     4981 gatgcagagt cacagttgga ggagagacaa aatttgcatg tgtcgatggt ccagcatttg
     5041 acgggcttaa agtagacttt gatgaagcaa tgagaaggca gaatatgtat aaagatatgg
     5101 agaggaaagt attagaaaat tatgagcacg aatgcaagtt agggggcatc ttaaatggct
     5161 aacatgtcgt taaaaaaagt accaatgcct gaacaggaac cagatcagcg gaacaaaaat
     5221 ttcaaagaag ttgcattagg gtatgaagaa aacatggctg ttgaagaagc cgaaagatgt
     5281 attcaatgta agaatcagcc atgtgttgaa ggttgccctg ttcacgtaaa aatacctgag
     5341 ttcatcaagc tgatagcaaa cagggacttt gaaggagcat atcaaaagat aaaagaaacg
     5401 aataatctac ctgcaatatg tggaagagta tgtccacagg aaagccagtg tgaatcagta
     5461 tgcacaagag gcaaaaaagg agaacctgta gctataggca gactggagag atttacagca
     5521 gattggcata tgaaaaataa tgaagataaa atagaaaaac cagagacgaa tggcagaaaa
     5581 gttgctgtta taggatctgg gcctgcagga ctatcgtgtg caggtgatct tgcaaaaatg
     5641 ggatatgaca caaccatctt tgaagcattt catacaccag gcggtgtttt aatgtacggt
     5701 atcccagagt ttagacttcc aaaagaaata gtgcaaaaag aaattgattc gttgaaaaaa
     5761 ttgggagtaa aaatagaaac caacatggtc ataggcaaga tacttacaat cgatgattta
     5821 ttcgacatgg ggtatgaagc agtgttcata gggacaggcg caggattgcc taaatttatg
     5881 aatatacctg gagaaaactt aaatggagtt tactcagcaa atgaattttt gacgaggatt
     5941 aatttgatga aagcttatga tttccctaac agtcccactc ctgtaaaagt aggcaagaaa
     6001 gttgctgttg ttggcggtgg aaacgtggcg atggatgcag caagatccgc aaagcgcatg
     6061 ggagcagaag aagtgtacat cgtatataga aggtcagagg aagaaatgcc tgcaaggctt
     6121 gaggaaatac accacgcaaa agaagaagga atcatattta agttattgac aaaccctgta
     6181 agaatcattg gtgatgaaag tggcagtgta aaaggaatcg aatgtgttaa catggtttta
     6241 ggcgatgtag acgaatcagg tagaagaaga cctgtagaag aaaaaggctc agagcatgta
     6301 atagatgtag atacagttat tatagcaata ggacaaagtc caaatccgct tataacatcc
     6361 acaacagaag gtttagaaaa gcaaagatgg ggtggaataa tagtcaacga agaaacattg
     6421 gagacaagcc gcagaggagt ttttgcgggc ggtgatgctg tgacaggtgc agctacagta
     6481 atacttgcaa tgggagctgg taaaaaagct gcagcttcta tacacaaata tttaagcgaa
     6541 aagtgataaa ggaggttata aaaatgtggg aaacaaaagt aaatcctagc aaaatttttg
     6601 aacttcgctg taaaaatacg acttattttg gtgttggcag tatacataag ataaaggata
     6661 tattagaaaa tctaaaaatt aatggcatta ataatgttat ttttataaca ggcaaaggtt
     6721 cgtataagac tagcggagct tgggatgttg taaggcctgt attggaagag cttgatttaa
     6781 aatattcact ttacgataaa gttggtccaa atcctaccgt tgatatgata gacgaagcgg
     6841 caaaaattgg acgagaatct ggagcgaaag ccgtaattgg aattggcggt ggtagcccta
     6901 tagatactgc aaaaagtgta gctgtgctat taaagtatac tgataaaaat gcacgagagc
     6961 tttataagca aaaatttatt cctgatgatg ctgttcctat aatagctata aatcttacac
     7021 atggtacagg aacagaggta gatagatttg cagtggctac tatacctgag aaaaattata
     7081 aaccagctat agcttatgat tgcttatatc ctatgtttgc tatagatgac ccttcgctta
     7141 tgacaaagct tgataaaaaa cagactatag ccgttactgt tgatgcttta aatcacatta
     7201 cagaagctgc tacgacactt gttgcatctc catatagtat tcttacggct aaggaaacag
     7261 taagattgat tgttcgatat cttcccgctg ctgtaaatga tcctttaaat atagtggcaa
     7321 gatattacct tctgtatgct tctgcattag ctggcatatc ttttgacaat ggtcttttgc
     7381 atcttactca tgctttagag catcctttaa gtgctgtaaa acctgaaata gctcatggtt
     7441 tagggcttgg tgcaatattg ccagcagtta taaaagctat ttatccagct acagcagaag
     7501 tattggctga tgtatatagt cctatagttc ctggtttaaa aggactgcct gttgaggcgg
     7561 agtatgtagc agaaaaagtt caagaatggc tttttagtgt ggggtgcatt caaaagttat
     7621 ctgattttgg gtttacgaaa gatgatattc ctaatcttgt aaaacttgct aaaacaactc
     7681 cttcccttga tgggttgtta tctattgcac ctgttgaagc tactgaatct gtcattgaaa
     7741 agatttattt aaaatctctt taattaatag aaataatttt gtttaacttt acaaacggga
//
