LOCUS       V004991                 6917 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004991
VERSION     V004991
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6917)
  AUTHORS   Lo J, Olson DG, Murphy SJ, Tian L, Hon S, Lanahan A, Guss AM, Lynd
            LR.
  TITLE     Engineering electron metabolism to increase ethanol production in
            Clostridium thermocellum
  JOURNAL   Metab. Eng. (2016) In press
   PUBMED   27989806
REFERENCE   2  (bases 1 to 6917)
  AUTHORS   Lo J, Olson DG, Murphy SJL., Tian L, Hon S, Lanahan AA, Guss AM,
            Lynd LR.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-NOV-2016) Thayer School of Engineering, Dartmouth
            College, 14 Engineering Drive, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 6917)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6917)
  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.
            (2016) In press"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (29-NOV-2016) Thayer School of Engineering, Dartmouth College, 14
            Engineering Drive, Hanover, NH 03755, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6917
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..209
                     /regulatory_class="promoter"
     CDS             210..1055
                     /codon_start=1
                     /gene="nfnA"
                     /product="NfnA"
                     /label="nfnA"
                     /protein_id="APQ46211.1"
                     /translation="MFKIVKKKILNPQVKLMEISAPHVAKKAEPGQFIILRVNENGERI
                     PLTIADFDREKGTVTIIFQEVGKTTKLLGTLEEGDELLDFVGPLGKASHFENVKKAAVI
                     GGGLGTAIAYPQAKKLHSMGVEVHTIAGFRNKDLIILEDEMRAVSSKLFITTDDGSNGN
                     KGFVSDVLKKLIEEGNKYDLVVAIGPLIMMKVISELTRPYGIKTIVSMNPVMIDGTGMC
                     GGCRVTVGGEIKFACVDGPDFDGHLVDFDEAMRRQAMYKKEESLALEKHNCKLGVEKNA
                     "
     gene            210..1055
                     /gene="nfnA"
                     /label="nfnA"
     CDS             1048..2442
                     /codon_start=1
                     /gene="nfnB"
                     /product="NfnB"
                     /label="nfnB"
                     /protein_id="APQ46210.1"
                     /translation="MPNMSPKKVPMPEQDPNVRIKNFLEVALGYTEQMAMEEAQRCLNC
                     KHKPCVSGCPVNVKIPEFVQLIAQGKFEKAYNKIRETNNLPAICGRVCPQENQCEKFCV
                     RGIKGEPVAIGRLERFAADWHMKNGTTSYEKPEKNGKRVAVIGSGPASLTCASDLAKLG
                     YEVTIFEAFHVPGGVLMYGIPEFRLPKKLVQEEIETIKQLGVEIKTNMVIGKVYSIDEL
                     KAEGYDAIFIGSGAGLPSFMKIPGENLNGVYSANEFLTRINLMKAYEFPNCDTPVKVGK
                     NVAVVGGGNVAMDAARSAKRLGAENVYIVYRRSEAEMPARLEEIHHAKEEGILFKFLTN
                     PTRILGTDDGWVKGMECIEMELGEPDESGRRRPVPKPGSEHVIDVETVIIAIGQTPNPL
                     IASTTPGLATQKWGGIIVDENTGATNIEGVYAGGDAVTGAATVILAMGAGKKAAKAIDE
                     YLKNKK"
     gene            1048..2442
                     /gene="nfnB"
                     /label="nfnB"
     CDS             2572..3219
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     CDS             complement(3303..4160)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(4161..4252)
                     /label="AmpR promoter"
     rep_origin      4852..5397
                     /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."
     rep_origin      5687..5867
     CDS             5868..6869
                     /label="repB"
                     /note="RepB replication protein"
ORIGIN
        1 gataaacaaa ggacggttca gggcttctgc tcatcctact ctgcattgta aaaaggtagg
       61 atgaattttt atttttaatc ttattgaaaa aaatttttga aaatcggttt tattaaaaaa
      121 aagtgggtat atttataata gtcaattgat tggttaaaaa aatttaaata agcaaacaga
      181 ataataacaa aagtaaggag gaatttgtta tgtttaaaat agttaaaaag aagattctca
      241 acccacaggt taagttgatg gaaatatcgg cgccgcacgt agctaaaaaa gcagagccgg
      301 gacagttcat aatcctgagg gtaaatgaaa atggtgaaag aattcccctc acaatagccg
      361 attttgaccg tgaaaaaggt acggtgacaa taatttttca ggaagtggga aaaaccacaa
      421 aactactggg aactttggaa gaaggagacg aattgctgga ctttgtcgga cctctgggta
      481 aagcatccca ctttgagaac gtaaaaaaag cggccgtcat tggaggaggc ttgggaacgg
      541 caatagcata tccccaggcc aaaaaacttc actccatggg tgttgaagtc catacgatag
      601 cgggtttcag aaacaaagac ctgataattc ttgaagatga aatgagagcc gtaagcagca
      661 aactttttat aaccaccgat gacggttcca acggcaataa aggtttcgta agcgacgtat
      721 taaagaaact tattgaagaa ggaaacaaat atgatttggt tgttgccatt gggccgctaa
      781 ttatgatgaa agttatcagc gagctcacaa gaccctacgg cattaaaacc attgtaagca
      841 tgaatccggt tatgattgac ggtaccggca tgtgcggagg ctgcagggtt accgttggcg
      901 gtgaaatcaa gttcgcatgt gtggacggac ctgattttga cgggcatctg gttgattttg
      961 acgaagcaat gagaaggcag gcaatgtata aaaaagaaga atccttggct ttggaaaaac
     1021 acaattgcaa attgggggtg gagaaaaatg cctaacatgt caccgaaaaa agttcccatg
     1081 ccggagcagg acccaaacgt cagaatcaaa aactttttgg aggttgcttt aggatatacc
     1141 gagcaaatgg caatggaaga agctcaaagg tgtcttaact gcaagcacaa accttgtgtt
     1201 tccggctgtc ccgtaaacgt aaaaattcct gagtttgtac agcttatcgc tcagggaaaa
     1261 tttgagaaag cctacaataa aataagagaa accaacaacc ttccggcaat atgcggcaga
     1321 gtctgtccgc aggaaaacca gtgtgaaaag ttctgtgtaa ggggtataaa aggtgaacct
     1381 gttgccatag gaaggcttga aagatttgcg gcggactggc acatgaaaaa cggcaccact
     1441 tcttatgaaa agcctgaaaa aaacggcaaa agggtggcag taataggttc gggacctgca
     1501 agccttacct gtgcaagcga cctggccaaa ctcggctacg aagtaacaat cttcgaagcc
     1561 tttcacgtgc ccggcggagt gttgatgtac ggtattccgg aattcaggct tccaaagaaa
     1621 ctggttcagg aggaaattga aaccataaag cagctgggtg tggaaattaa aacaaatatg
     1681 gttataggaa aggtttattc cattgacgaa ctcaaagctg aaggatatga tgccatattt
     1741 ataggctcgg gtgccggatt gccttcattt atgaaaattc ccggagaaaa cctcaacgga
     1801 gtttactcgg caaatgagtt tctcacaaga ataaacctca tgaaggctta tgaattcccc
     1861 aactgcgata ctcccgtgaa agtaggaaag aatgtcgccg ttgtgggcgg aggaaatgtc
     1921 gcaatggacg ccgcaagaag cgcaaaaaga cttggcgcgg aaaacgttta tatagtatac
     1981 aggcgttcgg aagcggaaat gcccgcaaga cttgaagaaa ttcatcacgc aaaggaagaa
     2041 ggaattttgt tcaaattcct tacaaacccc acaagaattc ttggcaccga cgacggctgg
     2101 gtcaaaggca tggagtgcat agagatggag ctgggcgaac ctgatgaatc cggaagaaga
     2161 agacccgtgc caaagccggg atccgaacat gtaattgatg ttgaaacggt tattatcgcc
     2221 atcggccaaa ctccaaatcc gttaattgcc tcaacaaccc cggggctggc cactcaaaaa
     2281 tggggcggaa ttattgtcga tgaaaacacc ggcgccacca acatagaagg tgtatatgcc
     2341 ggcggagatg cggtaaccgg tgccgcaacc gtcattcttg caatgggagc aggcaaaaaa
     2401 gccgcaaagg caattgacga atatcttaaa aacaaaaaat agttaataga aataattttg
     2461 tttaacttta caaacgggat tgacttttaa aaaaggattg attctaatga agaaagcaga
     2521 caagtaagcc tcctaaattc actttagata aaaatttagg aggcatatca aatgaacttt
     2581 aataaaattg atttagacaa ttggaagaga aaagagatat ttaatcatta tttgaaccaa
     2641 caaacgactt ttagtataac cacagaaatt gatattagtg ttttataccg aaacataaaa
     2701 caagaaggat ataaatttta ccctgcattt attttcttag tgacaagggt gataaactca
     2761 aatacagctt ttagaactgg ttacaatagc gacggagagt taggttattg ggataagtta
     2821 gagccacttt atacaatttt tgatggtgta tctaaaacat tctctggtat ttggactcct
     2881 gtaaagaatg acttcaaaga gttttatgat ttataccttt ctgatgtaga gaaatataat
     2941 ggttcgggga aattgtttcc caaaacacct atacctgaaa atgctttttc tctttctatt
     3001 attccatgga cttcatttac tgggtttaac ttaaatatca ataataatag taattacctt
     3061 ctacccatta ttacagcagg aaaattcatt aataaaggta attcaatata tttaccgcta
     3121 tctttacagg tacatcattc tgtttgtgat ggttatcatg caggattgtt tatgaactct
     3181 attcaggaat tgtcagatag gcctaatgac tggcttttat aatatgagat aatgccgact
     3241 gtacttttta cagtcggttt tctaatgtca ctagggctcg cctttgggaa gtttgaaggt
     3301 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3361 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3421 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     3481 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3541 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     3601 gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     3661 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     3721 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     3781 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     3841 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     3901 tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     3961 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4021 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4081 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4141 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4201 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aaagagtttg
     4261 aggaaaactt aagaaaacgg gaaactattt cctgttgaga atgtgaaatt aaatgataaa
     4321 aagtgatata atagggtcag gcgtacagaa gcctgacctt tttgcaaaat tgttttattg
     4381 gcgcggcgta tatatacagc atatatacaa tgtaaattta tacaatcaaa ttgttgatac
     4441 aatcgaatta tgcggctaaa aatttgctcg cgattgctga ataaaagata cgagagacct
     4501 ctcttgtatc ttttttattt tgagtggttt tgtccgttac actagaaaac cgaaagacaa
     4561 taaaaatttt attcttgctg agtctggctt tcggtaagct agacaaaacg gacaaaataa
     4621 aaattggcaa gggtttaaag gtggagattt tttgagtgat cttctcaaaa aatactacct
     4681 gtcccttgct gatttttaaa cgagcacgag agcaaaaccc ccctttgctg aggtggcaga
     4741 gggcaggttt ttttgtttct tttttctcgt aaaaaaaaga aaggtcttaa aggttttatg
     4801 gttttggtca tgagcctaga aatattttat ctgattaata agatgatctt cttgagatcg
     4861 ttttggtctg cgcgtaatct cttgctctga aaacgaaaaa accgccttgc agggcggttt
     4921 ttcgaaggtt ctctgagcta ccaactcttt gaaccgaggt aactggcttg gaggagcgca
     4981 gtcaccaaaa cttgtccttt cagtttagcc ttaaccggcg catgacttca agactaactc
     5041 ctctaaatca attaccagtg gctgctgcca gtggtgcttt tgcatgtctt tccgggttgg
     5101 actcaagacg atagttaccg gataaggcgc agcggtcgga ctgaacgggg ggttcgtgca
     5161 tacagtccag cttggagcga actgcctacc cggaactgag tgtcaggcgt ggaatgagac
     5221 aaacgcggcc ataacagcgg aatgacaccg gtaaaccgaa aggcaggaac aggagagcgc
     5281 acgagggagc cgccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     5341 cactgatttg agcgtcagat ttcgtgatgc ttgtcagggg ggcggagcct atggaaaaac
     5401 ggctttgccg cggccctctc acttccctgt taagtatctt cctggcatct tccaggaaat
     5461 ctccgccccg ttcgtaagcc atttccgctc gccgcagtcg aacgaccgag cgtagcgagt
     5521 cagtgagcga ggaagcggaa tatatcctgt atcacatatt ctgctgacgc accggtgcag
     5581 ccttttttct cctgccacat gaagcacttc actgacaccc tcatcagtgc caacatagta
     5641 agccagtata caaatgagtg ctttttttgc gttttgagcg tagcgaaaaa cgagttcttt
     5701 ctattcttga tacatataga aataacgtca tttttatttt agttgctgaa aggtgcgttg
     5761 aagtgttggt atgtatgtgt tttaaagtat tgaaaaccct taaaattggt tgcacagaaa
     5821 aaccccatct gttaaagtta taagtgacca aacaaataac taaatagatg ggggtttctt
     5881 ttaatattat gtgtcctaat agtagcattt attcagatga aaaatcaagg gttttagtgg
     5941 acaagacaaa aagtggaaaa gtgagaccat ggagagaaaa gaaaatcgct aatgttgatt
     6001 actttgaact tctgcatatt cttgaattta aaaaggctga aagagtaaaa gattgtgctg
     6061 aaatattaga gtataaacaa aatcgtgaaa caggcgaaag aaagttgtat cgagtgtggt
     6121 tttgtaaatc caggctttgt ccaatgtgca actggaggag agcaatgaaa catggcattc
     6181 agtcacaaaa ggttgttgct gaagttatta aacaaaagcc aacagttcgt tggttgtttc
     6241 tcacattaac agttaaaaat gtttatgatg gcgaagaatt aaataagagt ttgtcagata
     6301 tggctcaagg atttcgccga atgatgcaat ataaaaaaat taataaaaat cttgttggtt
     6361 ttatgcgtgc aacggaagtg acaataaata ataaagataa ttcttataat cagcacatgc
     6421 atgtattggt atgtgtggaa ccaacttatt ttaagaatac agaaaactac gtgaatcaaa
     6481 aacaatggat tcaattttgg aaaaaggcaa tgaaattaga ctatgatcca aatgtaaaag
     6541 ttcaaatgat tcgaccgaaa aataaatata aatcggatat acaatcggca attgacgaaa
     6601 ctgcaaaata tcctgtaaag gatacggatt ttatgaccga tgatgaagaa aagaatttga
     6661 aacgtttgtc tgatttggag gaaggtttac accgtaaaag gttaatctcc tatggtggtt
     6721 tgttaaaaga aatacataaa aaattaaacc ttgatgacac agaagaaggc gatttgattc
     6781 atacagatga tgacgaaaaa gccgatgaag atggattttc tattattgca atgtggaatt
     6841 gggaacggaa aaattatttt attaaagagt agttcaaccc ggggatcctc tagagtcgac
     6901 ctgcaggcat gcaagct
//
