LOCUS       V003253                 6924 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003253
VERSION     V003253
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6924)
  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 6924)
  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 6924)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6924)
  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..6924
                     /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..6887
                     /codon_start=1
                     /gene="adhE"
                     /product="AdhE"
                     /label="adhE"
                     /note="bifunctional alcohol and aldehyde dehydrogenase"
                     /protein_id="ASK86055.1"
                     /translation="MATTKTELDVQKQIDLLVSRAQEAQKKFMSYTQEQIDAIVKAMAL
                     AGVDKHVELAKMAYEETKMGVYEDKITKNLFATEYVYHDIKNEKTVGIINENIEENYME
                     VAEPIGVIAGVTPVTNPTSTTMFKCLISIKTRNPIIFSFHPKAIKCSIAAAKVMYEAAL
                     KAGAPEGCIGWIETPSIEATQLLMTHPGVSLILATGGAGMVKAAYSSGKPALGVGPGNV
                     PCYIEKSANIKRAVSDLILSKTFDNGVICASEQAVIIDEEIADEVKKLMKEYGCYFLNK
                     DEIKKLEKFAIDEQSCAMSPAVVGQPAAKIAEMAGFKVPEGTKILVAEYEGVGPKYPLS
                     REKLSPILACYTVKDYNEGIKKCEEMTEFGGLGHSAVIHSENQNVINEFARRVRTGRLI
                     VNSPSSQGAIGDIYNTNTPSLTLGCGSMGRNSTTDNVSVKNLLNIKRVVIRKDRMKWFK
                     IPPKIYFESGSLQYLCKVKRKKAFIVTDPFMVKLGFVDKVTYQLDKANIEYEIFSEVEP
                     DPSVDTVMNGVKIMNSYNPDLIIAVGDGSAIDAAKGMWLFYEYPDTEFETLRLKFADIR
                     KRAFKFPELGKKALFIAIPTTSGTGSEVTAFAVITDKKRNIKYPLADYELTPDIAIIDP
                     DLTKTVPPSVTADTGMDVLTHAIEAYVSVMASDYTDALAEKAIKIVFEYLPRAYKNGND
                     EEAREKMHNASCMAGMAFTNAFLGINHSMAHILGGKFHIPHGRANAILLPYVIRYNAEK
                     PTKFVAFPQYEYPKAAERYAEIAKFLGLPASTVEEGVESLIEAIKNLMKELNIPLTLKD
                     AGINKEQFEKEIEEMSDIAFNDQCTGTNPRMPLTKEIAEIYRKAYGA"
     gene            4308..6887
                     /gene="adhE"
                     /label="adhE"
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 gcaacgacaa
     4321 aaacggaatt agacgttcag aagcagatag atctacttgt gtcaagagca caagaggctc
     4381 agaaaaaatt catgtcttac acgcaagagc aaatcgacgc aatagttaag gcaatggctt
     4441 tagcaggcgt tgacaaacac gtagagctgg caaagatggc gtacgaagag acaaaaatgg
     4501 gtgtatacga agataagata acaaaaaatc tcttcgcaac agagtacgtg taccacgaca
     4561 taaaaaatga aaagactgta ggaatcataa acgagaacat agaagaaaac tacatggaag
     4621 tggcagaacc gataggcgta attgccggtg tcacacctgt cacaaaccca acatctacca
     4681 cgatgtttaa atgcttaata tccataaaga cgcgaaatcc tataatattc agcttccatc
     4741 caaaggcaat aaagtgcagc atcgcagcag ccaaagtgat gtatgaagct gcactaaagg
     4801 caggcgcacc tgaaggatgc ataggatgga tagaaacgcc atcaattgag gccacacagc
     4861 ttctcatgac acatccaggc gtatcgctga tccttgcaac gggcggtgca ggaatggtaa
     4921 aagcggcata cagctcagga aaaccggcat taggcgtagg tcctggcaat gtgccatgct
     4981 acatcgaaaa atcagcaaac ataaagaggg ctgtatcgga tctcatacta agcaagacat
     5041 ttgacaatgg agtaatatgc gcatcagagc aggccgtaat aatagacgag gaaatagcag
     5101 atgaagtcaa aaagcttatg aaagaatacg gctgctactt cttaaacaaa gatgaaataa
     5161 agaagcttga gaaatttgca attgatgagc aaagctgcgc catgagccct gcagtggtag
     5221 gtcagccagc ggcgaagatt gctgaaatgg caggcttcaa agtccccgaa ggcacaaaga
     5281 tattagtggc agagtacgaa ggagtaggtc caaaatatcc tctatcaagg gagaaactaa
     5341 gcccgattct tgcttgctac accgtcaaag actacaatga aggaatcaaa aagtgcgagg
     5401 aaatgactga attcggaggt ttaggccact ctgctgtaat acactctgaa aatcaaaacg
     5461 tcataaatga atttgcaagg cgagtccgca caggaagact tatcgtaaat tcaccatcat
     5521 cacagggagc aataggagat atatacaata caaacacgcc atcacttaca ttaggctgtg
     5581 gttctatggg aagaaactca acgacagaca atgtaagcgt caagaacctt ttgaatatta
     5641 agcgtgtcgt gataaggaag gatagaatga aatggttcaa gattccaccg aagatttact
     5701 ttgaaagcgg gtcactccag tacctgtgca aagtcaaaag aaaaaaagcg tttatcgtca
     5761 cagatccatt catggttaag cttggcttcg tagacaaagt gacatatcaa ttagacaaag
     5821 caaacatcga atacgaaata ttctcagaag tagagccaga tccatctgtt gacacagtca
     5881 tgaacggcgt aaaaataatg aattcgtaca atcctgactt aataatcgct gtaggcgatg
     5941 gctctgcaat agacgcagca aagggaatgt ggcttttcta cgaatatcct gatacagagt
     6001 ttgaaacatt gaggcttaaa tttgcagaca tcagaaaaag ggcatttaag ttcccagaac
     6061 ttggcaaaaa agcgctattc atcgcaatac cgacaacaag cggcacaggc tcagaagtga
     6121 cagcatttgc cgtaataacc gacaaaaaga gaaacatcaa gtatccactg gcagactacg
     6181 aacttacacc tgacatagcc ataatagatc ctgaccttac aaagactgta ccgccatctg
     6241 taacagcaga cacaggcatg gatgtgctga cacacgccat agaagcatac gtatcagtaa
     6301 tggcatcaga ctacacagat gcactggcgg aaaaggctat aaagatcgta tttgaatacc
     6361 tgccaagggc ttataaaaac ggcaatgatg aagaagcccg cgaaaagatg cacaatgctt
     6421 cctgcatggc tggtatggca ttcacaaatg cattcttagg aataaaccac agcatggcac
     6481 acatactggg cggaaagttc cacataccac acggaagagc aaatgcaata cttctgccgt
     6541 atgtaataag gtacaatgca gaaaaaccta caaagtttgt ggcattccca caatacgaat
     6601 atccaaaagc agcagaaaga tatgcggaaa tcgccaaatt cttaggactg cctgcttcaa
     6661 ctgttgaaga aggcgtagaa agcttaatag aagctataaa gaacctcatg aaagagctta
     6721 acattccgct tacacttaaa gacgccggca tcaacaaaga acagtttgaa aaagaaatag
     6781 aggaaatgtc agacatcgcc ttcaacgatc agtgcacagg gacaaacccg agaatgcctc
     6841 tcacaaaaga aattgcagag atctacagaa aagcatacgg tgcatagtta atagaaataa
     6901 ttttgtttaa ctttacaaac ggga
//
