LOCUS       V003140                 8732 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003140
VERSION     V003140
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8732)
  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 8732)
  AUTHORS   Lo J, Olson DG, Murphy SJ, Tian L, Hon S, Lanahan A, Guss AM, Lynd
            LR.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-DEC-2016) Thayer School of Engineering, Dartmouth
            College, 14 Engineering Drive, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 8732)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8732)
  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
            (05-DEC-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..8732
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      33..221
                     /label="CYC1 terminator"
                     /note="transcription terminator for CYC1"
     rep_origin      complement(481..1069)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(1156..1734)
                     /codon_start=1
                     /product="Tdk"
                     /EC_number="2.7.1.21"
                     /label="Tdk"
                     /note="thymidine kinase"
                     /protein_id="APQ46124.1"
                     /translation="MYGPKDHGYIEVVTGPMFSGKSEELIRRIKRAKIARQKVQVFKPA
                     IDDRYSIDKVVSHNGDNMHAIAIVKASDILAYAEEDTDVFAIDEVQFFDSEIVDIVKEI
                     ADSGKRVICAGLDMDFRGEPFGPTPELMAIAEFVDKLTAICMKCGNPATRTQRLINGKP
                     ANYDDPIIMVGAKESYEARCRKCHEVPRT"
     CDS             complement(2434..3435)
                     /label="repB"
                     /note="RepB replication protein"
     CDS             complement(3996..4853)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(4854..4958)
                     /label="AmpR promoter"
     CDS             6126..6773
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     CDS             6794..7339
                     /codon_start=1
                     /product="Hpt"
                     /EC_number="2.4.2.8"
                     /label="Hpt"
                     /note="hypoxanthine phosphoribosyltransferase"
                     /protein_id="APQ46123.1"
                     /translation="MENLSKDIDEILITEEELKEKIKELGRQITKDYKGKNLMLVGVLK
                     GALMFMADLSRHIDLPLSLDFMAVSSYGSSTHSSGIVKIIKDLDISIEGKDVLIVEDII
                     DSGLTLSYLRETLLGRKPKSLKICTILDKPERREASVKVDYVGFKIPDKFVVGYGLDFD
                     EKYRNLPFIGVLKPEMYS"
ORIGIN
        1 ctgctaatag tagtgaaaaa atcagggata atgccgctct aaccgaaaag gaaggagtta
       61 gacaacctga agtctaggtc cctatttatt tttttatagt tatgttagta ttaagaacgt
      121 tatttatatt tcaaattttt cttttttttc tgtacagacg cgtgtacgca tgtaacatta
      181 tactgaaaac cttgcttgag aaggttttgg gacgctcgaa ggctttaatt tgcaagctgc
      241 ggccctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
      301 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
      361 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
      421 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
      481 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
      541 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
      601 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
      661 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
      721 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
      781 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
      841 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
      901 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
      961 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     1021 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     1081 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     1141 catgaaaaaa tccgcttaag tccgcgggac ttcatggcac tttctacacc ttgcttcata
     1201 agactccttt gctccaacca ttataatggg atcgtcgtaa ttggcaggct tcccatttat
     1261 aagcctttgc gtgcgagtag caggattgcc acacttcatg catatagcag taagcttatc
     1321 gacaaattca gctatggcca tcaattctgg agtcggacca aatggttcac ctctaaagtc
     1381 catgtcaagc cctgcacata tgactctttt cccgctatcg gcaatctctt ttacgatgtc
     1441 gactatttca gaatcaaaaa attgaacttc atctatagca aatacatccg tatcttcttc
     1501 agcataagcc aatatgtcag aagcctttac tatggcaatg gcgtgcatgt tgtcgccgtt
     1561 atgagatacg actttgtcta tggaatacct atcgtctata gccggtttga aaacctgaac
     1621 tttttgcctg gcaatcttag ctctctttat ccttcttata agttcctcac tttttccact
     1681 gaacatggga ccagttacaa cttctatgta cccgtggtct ttaggcccat acatctttcg
     1741 tcctccttaa aattttcgtt tatttgtttt ttatttagtg tgcctaaaag caccatctta
     1801 tacattcgta ccttatatct cttatcttgt atctcttacc tcatatctct tattattaac
     1861 taccgaagta tattttcata acggaatctt atgacttatt tttctgactg ccgttttacc
     1921 atacggttaa tttatacgga aacagctaat ctaaccttgc tgaaggcatg tttaccaaaa
     1981 tattatatct actttaccat attaattcaa gtggtcattt tgatatttta cgccaaaatt
     2041 tttagcgttc ctttacttga taatttatcc aaaaaccttc attcgtcaaa acaattatga
     2101 tataattaat ttatgatatg attaattaat atttacggat aactaatcat ttcaatgcaa
     2161 tagatcaaca tgcaaatcaa tatccacctt actccgaaca ttttgcacca acgaatgccc
     2221 agcactatat cccggcaagc aatataacgt ctataatttt ttatgataac gataaaatta
     2281 aattttatga aaggggtaaa agcatgagaa aaatagctgt attgttaatt actttgacgt
     2341 tcttagtcac gactcgaatt taggaggctt acttgtctgc tttcttcatt agaatcaatc
     2401 cttttttaaa agtcaatccc gtttgttgaa ctactcttta ataaaataat ttttccgttc
     2461 ccaattccac attgcaataa tagaaaatcc atcttcatcg gctttttcgt catcatctgt
     2521 atgaatcaaa tcgccttctt ctgtgtcatc aaggtttaat tttttatgta tttcttttaa
     2581 caaaccacca taggagatta accttttacg gtgtaaacct tcctccaaat cagacaaacg
     2641 tttcaaattc ttttcttcat catcggtcat aaaatccgta tcctttacag gatattttgc
     2701 agtttcgtca attgccgatt gtatatccga tttatattta tttttcggtc gaatcatttg
     2761 aacttttaca tttggatcat agtctaattt cattgccttt ttccaaaatt gaatccattg
     2821 tttttgattc acgtagtttt ctgtattctt aaaataagtt ggttccacac ataccaatac
     2881 atgcatgtgc tgattataag aattatcttt attatttatt gtcacttccg ttgcacgcat
     2941 aaaaccaaca agatttttat taattttttt atattgcatc attcggcgaa atccttgagc
     3001 catatctgac aaactcttat ttaattcttc gccatcataa acatttttaa ctgttaatgt
     3061 gagaaacaac caacgaactg ttggcttttg tttaataact tcagcaacaa ccttttgtga
     3121 ctgaatgcca tgtttcattg ctctcctcca gttgcacatt ggacaaagcc tggatttaca
     3181 aaaccacact cgatacaact ttctttcgcc tgtttcacga ttttgtttat actctaatat
     3241 ttcagcacaa tcttttactc tttcagcctt tttaaattca agaatatgca gaagttcaaa
     3301 gtaatcaaca ttagcgattt tcttttctct ccatggtctc acttttccac tttttgtctt
     3361 gtccactaaa acccttgatt tttcatctga ataaatgcta ctattaggac acataatatt
     3421 aaaagaaacc cccatctatt tagttatttg tttggtcact tataacttta acagatgggg
     3481 tttttctgtg caaccaattt taagggtttt caatacttta aaacacatac ataccaacac
     3541 ttcaacgcac ctttcagcaa ctaaaataaa aatgacgtta tttctatatg tatcaagaat
     3601 agaaagaact cgtttttcgc tacgctcaaa acgcaaaaaa agcactcatt cgagtgcttt
     3661 ttcttatcgc tccaaatcat gcgatttttt cctctttgct tttctttgct cacgaagttc
     3721 tcgatcacgc tgcaaaacat cttgaagcga aaaagtattc ttcttttctt ccgatcgctc
     3781 atgctgacgc acgaaaagcc ctctaggcgc ataggaacaa ctcctaaatg catgtgaggg
     3841 gttttctcgt ccatgtgaac agtcgcatac gcaatatttt gtttcccata gcggccgcga
     3901 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     3961 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     4021 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     4081 actacgatac gggagcgctt accatctggc cccagtgctg caatgatacc gcgagaccca
     4141 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     4201 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     4261 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     4321 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     4381 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     4441 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     4501 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     4561 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     4621 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     4681 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     4741 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     4801 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     4861 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     4921 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     4981 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     5041 aggcgataaa actgatacga atgaatatcg tgattacagt tgtttattga aagacagcat
     5101 aaattagttg ctaaaaattg gttactaaaa gatattttaa tttggcaaat tcttatttga
     5161 cttatttaca agttgcaaat ataatattca gtggaaatat atttagagga agaacatatt
     5221 cagaaaaaag cagcctatcg aaaaaacaaa aaagcctgtc aatggggatg ggttttatca
     5281 ctctagtcga acagcttgct acagaaattg ctcattttta gttggtgttt cgtgatattc
     5341 tatgttgtat acaatatacc gccagaaatc acagaaatcc agtaattttg catggtagta
     5401 acaattgcct gatttgtaca aatcaaaata ttttaataaa attgccgatg tgctattata
     5461 aaaaagtgag taattaaaat aggcatatgg gtggtatata aaattttact gtggaatata
     5521 aaaattattt aaaatggggt tatacgaatc gtgggaatag gcatggaaaa gatttttttg
     5581 cccgaggatg cattgataaa gtatttcttt tccgaaagag aggaaaagat tctaaagagt
     5641 tttggaaata ctgatgaata ttgtgtgcag agtacaattc tatggacaag aaaagaggct
     5701 ttgtcaaaac tttttcgtct gggaatgagg atggatttta aaaagctgga tactttggag
     5761 ggcgaggtgg tttttcagga aacaaacagg gcgcgtctgt tttcttttat atgcaataat
     5821 tactgtatct ctctggcatt gccaggtttt aataaagatt aaaattattg actagaaata
     5881 aaaaaattgt ccataatatt aatggacaaa aaaacaaaga attacatcaa aggaagataa
     5941 aaatactttg ttaaaaaatt aattattttt tatctaaact attgaaaatg aaaataaaat
     6001 aatataaaat gaatcatagt gcaagagata cttgccagag gatgaatatt ttactgcatt
     6061 catgctttat ggcagctaat agaggcatta aattaaattt taatttacaa taggaggcga
     6121 tattaatgaa ctttaataaa attgatttag acaattggaa gagaaaagag atatttaatc
     6181 attatttgaa ccaacaaacg acttttagta taaccacaga aattgatatt agtgttttat
     6241 accgaaacat aaaacaagaa ggatataaat tttaccctgc atttattttc ttagtgacaa
     6301 gggtgataaa ctcaaataca gcttttagaa ctggttacaa tagcgacgga gagttaggtt
     6361 attgggataa gttagagcca ctttatacaa tttttgatgg tgtatctaaa acattctctg
     6421 gtatttggac tcctgtaaag aatgacttca aagagtttta tgatttatac ctttctgatg
     6481 tagagaaata taatggttcg gggaaattgt ttcccaaaac acctatacct gaaaatgctt
     6541 tttctctttc tattattcca tggacttcat ttactgggtt taacttaaat atcaataata
     6601 atagtaatta ccttctaccc attattacag caggaaaatt cattaataaa ggtaattcaa
     6661 tatatttacc gctatcttta caggtacatc attctgtttg tgatggttat catgcaggat
     6721 tgtttatgaa ctctattcag gaattgtcag ataggcctaa tgactggctt ttataataaa
     6781 ggaggtcgac gtcatggaaa atttatcaaa agacatcgat gaaattttga tcacagaaga
     6841 agaacttaag gaaaagataa aagagcttgg gaggcaaatc acaaaagact acaaagggaa
     6901 aaatttgatg ttggtaggag ttttaaaagg tgctttaatg tttatggctg atttgtcaag
     6961 acacatagat ttgcctttat cacttgattt tatggctgtt tccagctatg gaagctcaac
     7021 tcattcatca ggaatagtaa agataatcaa agatcttgat ataagcatag aaggcaaaga
     7081 tgttctgatt gtggaagaca taattgacag cggtttgact ttgtcttact taagggaaac
     7141 tttacttgga aggaagccaa aaagcctgaa aatatgcaca atattagaca aaccggagag
     7201 aagagaagca tctgtaaaag tcgattatgt aggatttaag atacctgata agtttgtcgt
     7261 gggttatgga ttggactttg atgaaaagta caggaacctt ccttttatag gcgttttgaa
     7321 acctgaaatg tacagctaag cccgggcctc gagaaaacaa aaggctcagt cggaagactg
     7381 ggccttttgt tttggtaccg aattcggcgc gcctcagcgt ttaaacccgc tgatcctaga
     7441 gggccgcatc atgtaattag ttatgtcacg cttacattca cgccctcccc atgctttaat
     7501 acatacctgg cccagtagtt cagaaaactg atacgaatga atatcgtgat tacagttgtt
     7561 tattgaaaga cagcataaat tagttgctaa aaattggtta ctaaaagata ttttaatttg
     7621 gcaaattctt atttgactta tttacaagtt gcaaatataa tattcagtgg aaatatattt
     7681 agaggaagaa catattcaga aaaaagcagc ctatcgaaaa aacaaaaaag cctgtcaatg
     7741 gggatgggtt ttatcactct agtcgaacag cttgctacag aaattgctca tttttagttg
     7801 gtgtttcgtg atattctatg ttgtatacaa tataccgcca gaaatcacag aaatccagta
     7861 attttgcatg gtagtaacaa ttgcctgatt tgtacaaatc aaaatatttt aataaaattg
     7921 ccgatgtgct attataaaaa agtgagtaat taaaataggc atatgggtgg tatataaaat
     7981 tttactgtgg aatataaaaa ttatttaaaa tggggttata cgagataaac aaaggacggt
     8041 tcagggcttc tgctcatcct actctgcatt gtaaaaaggt aggatgaatt tttattttta
     8101 atcttattga aaaaaatttt tgaaaatcgg ttttattaaa aaaaagtggg tatatttata
     8161 atagtcaatt gattggttaa aaaaatttaa ataagcaaac agaataataa caaaagtaag
     8221 gaggaatttg ttatgtttgc taggaatttc ttcaaaggcg gaacgaaagt tcctcacttc
     8281 aaaaatactg ctgaatgtga aactgtcaaa atgggggtcc ctgaaaaggt ggtaattccc
     8341 atgcttcagc atgtcggagc gccgtgtgaa ccctgtgtaa agaaagggga tactgtccgg
     8401 gtgggacagg tgataggaaa ttctgataag tatatttcgt cacctatcca ttcaagcgta
     8461 tccggaacgg taaacgatgt ggctccgatg ctttttgccg gcgggattta cgttaccgcg
     8521 gtggaaatta agactgacgg aaaacaggaa atttatgaag gtatcaagcc tccgagtttt
     8581 tcaaacagca aagagtttat ttcagccatc cgggaatcgg gcctcgtagg acttggcggt
     8641 gctggatttc cggctcatgt caagctttca ccccctccgg acaaaaagat tgacacgctg
     8701 ataatcaatg cggctgagtg tgaaccttat at
//
