LOCUS       V005307                 8515 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V005307
VERSION     V005307
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8515)
  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 8515)
  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 8515)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8515)
  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..8515
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(561..1418)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(1419..1523)
                     /label="AmpR promoter"
     CDS             3193..3840
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     CDS             3861..4406
                     /codon_start=1
                     /product="Hpt"
                     /EC_number="2.4.2.8"
                     /label="Hpt"
                     /note="hypoxanthine phosphoribosyltransferase"
                     /protein_id="APQ46113.1"
                     /translation="MENLSKDIDEILITEEELKEKIKELGRQITKDYKGKNLMLVGVLK
                     GALMFMADLSRHIDLPLSLDFMAVSSYGSSTHSSGIVKIIKDLDISIEGKDVLIVEDII
                     DSGLTLSYLRETLLGRKPKSLKICTILDKPERREASVKVDYVGFKIPDKFVVGYGLDFD
                     EKYRNLPFIGVLKPEMYS"
     terminator      5113..5301
                     /label="CYC1 terminator"
                     /note="transcription terminator for CYC1"
     rep_origin      complement(5561..6149)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6236..6814)
                     /codon_start=1
                     /product="Tdk"
                     /EC_number="2.7.1.21"
                     /label="Tdk"
                     /note="thymidine kinase"
                     /protein_id="APQ46114.1"
                     /translation="MYGPKDHGYIEVVTGPMFSGKSEELIRRIKRAKIARQKVQVFKPA
                     IDDRYSIDKVVSHNGDNMHAIAIVKASDILAYAEEDTDVFAIDEVQFFDSEIVDIVKEI
                     ADSGKRVICAGLDMDFRGEPFGPTPELMAIAEFVDKLTAICMKCGNPATRTQRLINGKP
                     ANYDDPIIMVGAKESYEARCRKCHEVPRT"
     CDS             complement(7514..8515)
                     /label="repB"
                     /note="RepB replication protein"
ORIGIN
        1 ctatttagtt atttgtttgg tcacttataa ctttaacaga tggggttttt ctgtgcaacc
       61 aattttaagg gttttcaata ctttaaaaca catacatacc aacacttcaa cgcacctttc
      121 agcaactaaa ataaaaatga cgttatttct atatgtatca agaatagaaa gaactcgttt
      181 ttcgctacgc tcaaaacgca aaaaaagcac tcattcgagt gctttttctt atcgctccaa
      241 atcatgcgat tttttcctct ttgcttttct ttgctcacga agttctcgat cacgctgcaa
      301 aacatcttga agcgaaaaag tattcttctt ttcttccgat cgctcatgct gacgcacgaa
      361 aagccctcta ggcgcatagg aacaactcct aaatgcatgt gaggggtttt ctcgtccatg
      421 tgaacagtcg catacgcaat attttgtttc ccatagcggc cgcgattatc aaaaaggatc
      481 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
      541 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
      601 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
      661 cgcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
      721 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
      781 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
      841 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
      901 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
      961 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     1021 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     1081 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     1141 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     1201 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     1261 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     1321 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     1381 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     1441 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     1501 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     1561 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcg attatcaaga
     1621 ccggaatgcc ccttataaag aaaagggtca cggtggacgg accttcggta aaaagaccgg
     1681 gcaatgtgga agttttgata ggtactccca tatcggaggt tttcagcttt tgcggaggtt
     1741 ttggcattgt tcccaaaaag atactgatgg gtggaccaat gatgggaata gcgcagtata
     1801 cgctggatac accggttctc aagcatacaa atgcccttct ggcttttgat gaaacaactg
     1861 ccgtattgcc aaaggagcaa gcttgtataa ggtgcggaag gtgtgttagg gcatgtccga
     1921 tgaatctttt accactttat cttaacaata attcgataag gggaaatatt gaagagctta
     1981 acagatacca tgtaatggat tgcatagaat gtggcagctg tgcttatgta tgtccggcaa
     2041 aaattaatct tgtccagtca ataaggcttg gcaaggcaca ggtaagacaa gcggcagcga
     2101 aagggggaaa ttaaatgagt ggttttaaga ctgtaacgaa agggataata aaggaaaacc
     2161 cggtattcag gcttgtcctt ggtacatgcc ccactttggc tgtgacaaca tcagccgaaa
     2221 atgccttggg aatgggggcg gccgttatat ttgtgttgat ggggtcaaat gtagtgattt
     2281 cccttttgag aaaggtgata cctgacaagg tcagaattcc tgcatatata acaattattt
     2341 cgggatttgt gacgatagtt cagatgctga ttaaggcata tcttcctgct attgataagc
     2401 agctgggtat atttatacct cttatagttg taaactgtat catacttgca agagcggaga
     2461 tgtttgcaag caagaacggt ccactcatgt caattcttga cgggcttggc atgggaatag
     2521 gttttacctt gtcgcttctt gttatgggaa gcgtcagaga gatacttgga aacggcacat
     2581 ggtttggaat gaatattacc ggtggaattt atgaatcgtg ggaataggca tggaaaagat
     2641 ttttttgccc gaggatgcat tgataaagta tttcttttcc gaaagagagg aaaagattct
     2701 aaagagtttt ggaaatactg atgaatattg tgtgcagagt acaattctat ggacaagaaa
     2761 agaggctttg tcaaaacttt ttcgtctggg aatgaggatg gattttaaaa agctggatac
     2821 tttggagggc gaggtggttt ttcaggaaac aaacagggcg cgtctgtttt cttttatatg
     2881 caataattac tgtatctctc tggcattgcc aggttttaat aaagattaaa attattgact
     2941 agaaataaaa aaattgtcca taatattaat ggacaaaaaa acaaagaatt acatcaaagg
     3001 aagataaaaa tactttgtta aaaaattaat tattttttat ctaaactatt gaaaatgaaa
     3061 ataaaataat ataaaatgaa tcatagtgca agagatactt gccagaggat gaatatttta
     3121 ctgcattcat gctttatggc agctaataga ggcattaaat taaattttaa tttacaatag
     3181 gaggcgatat taatgaactt taataaaatt gatttagaca attggaagag aaaagagata
     3241 tttaatcatt atttgaacca acaaacgact tttagtataa ccacagaaat tgatattagt
     3301 gttttatacc gaaacataaa acaagaagga tataaatttt accctgcatt tattttctta
     3361 gtgacaaggg tgataaactc aaatacagct tttagaactg gttacaatag cgacggagag
     3421 ttaggttatt gggataagtt agagccactt tatacaattt ttgatggtgt atctaaaaca
     3481 ttctctggta tttggactcc tgtaaagaat gacttcaaag agttttatga tttatacctt
     3541 tctgatgtag agaaatataa tggttcgggg aaattgtttc ccaaaacacc tatacctgaa
     3601 aatgcttttt ctctttctat tattccatgg acttcattta ctgggtttaa cttaaatatc
     3661 aataataata gtaattacct tctacccatt attacagcag gaaaattcat taataaaggt
     3721 aattcaatat atttaccgct atctttacag gtacatcatt ctgtttgtga tggttatcat
     3781 gcaggattgt ttatgaactc tattcaggaa ttgtcagata ggcctaatga ctggctttta
     3841 taataaagga ggtcgacgtc atggaaaatt tatcaaaaga catcgatgaa attttgatca
     3901 cagaagaaga acttaaggaa aagataaaag agcttgggag gcaaatcaca aaagactaca
     3961 aagggaaaaa tttgatgttg gtaggagttt taaaaggtgc tttaatgttt atggctgatt
     4021 tgtcaagaca catagatttg cctttatcac ttgattttat ggctgtttcc agctatggaa
     4081 gctcaactca ttcatcagga atagtaaaga taatcaaaga tcttgatata agcatagaag
     4141 gcaaagatgt tctgattgtg gaagacataa ttgacagcgg tttgactttg tcttacttaa
     4201 gggaaacttt acttggaagg aagccaaaaa gcctgaaaat atgcacaata ttagacaaac
     4261 cggagagaag agaagcatct gtaaaagtcg attatgtagg atttaagata cctgataagt
     4321 ttgtcgtggg ttatggattg gactttgatg aaaagtacag gaaccttcct tttataggcg
     4381 ttttgaaacc tgaaatgtac agctaagccc gggcctcgag aaaacaaaag gctcagtcgg
     4441 aagactgggc cttttgtttt ggtaccgaat tcggcgcgcc tcagcgttta aacccgctga
     4501 tcctagaggg ccgcatcatg taattagtta tgtcacgctt acattcacgc cctccccatg
     4561 ctttaataca tacctggccc agtagttcag atgagtggtt ttaagactgt aacgaaaggg
     4621 ataataaagg aaaacccggt attcaggctt gtccttggta catgccccac tttggctgtg
     4681 acaacatcag ccgaaaatgc cttgggaatg ggggcggccg ttatatttgt gttgatgggg
     4741 tcaaatgtag tgatttccct tttgagaaag gtgatacctg acaaggtcag aattcctgca
     4801 tatataacaa ttatttcggg atttgtgacg atagttcaga tgctgattaa ggcatatctt
     4861 cctgctattg ataagcagct gggtatattt atacctctta tagttgtaaa ctgtatcata
     4921 cttgcaagag cggagatgtt tgcaagcaag aacggtccac tcatgtcaat tcttgacggg
     4981 cttggcatgg gaataggttt taccttgtcg cttcttgtta tgggaagcgt cagagagata
     5041 cttggaaacg gcacatggtt tggaatgaat attaccggtg ctgctaatag tagtgaaaaa
     5101 atcagggata atgccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc
     5161 cctatttatt tttttatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt
     5221 cttttttttc tgtacagacg cgtgtacgca tgtaacatta tactgaaaac cttgcttgag
     5281 aaggttttgg gacgctcgaa ggctttaatt tgcaagctgc ggccctgcat taatgaatcg
     5341 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5401 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5461 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5521 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5581 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5641 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5701 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5761 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5821 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5881 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5941 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6001 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6061 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6121 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6181 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgaaaaaa tccgcttaag
     6241 tccgcgggac ttcatggcac tttctacacc ttgcttcata agactccttt gctccaacca
     6301 ttataatggg atcgtcgtaa ttggcaggct tcccatttat aagcctttgc gtgcgagtag
     6361 caggattgcc acacttcatg catatagcag taagcttatc gacaaattca gctatggcca
     6421 tcaattctgg agtcggacca aatggttcac ctctaaagtc catgtcaagc cctgcacata
     6481 tgactctttt cccgctatcg gcaatctctt ttacgatgtc gactatttca gaatcaaaaa
     6541 attgaacttc atctatagca aatacatccg tatcttcttc agcataagcc aatatgtcag
     6601 aagcctttac tatggcaatg gcgtgcatgt tgtcgccgtt atgagatacg actttgtcta
     6661 tggaatacct atcgtctata gccggtttga aaacctgaac tttttgcctg gcaatcttag
     6721 ctctctttat ccttcttata agttcctcac tttttccact gaacatggga ccagttacaa
     6781 cttctatgta cccgtggtct ttaggcccat acatctttcg tcctccttaa aattttcgtt
     6841 tatttgtttt ttatttagtg tgcctaaaag caccatctta tacattcgta ccttatatct
     6901 cttatcttgt atctcttacc tcatatctct tattattaac taccgaagta tattttcata
     6961 acggaatctt atgacttatt tttctgactg ccgttttacc atacggttaa tttatacgga
     7021 aacagctaat ctaaccttgc tgaaggcatg tttaccaaaa tattatatct actttaccat
     7081 attaattcaa gtggtcattt tgatatttta cgccaaaatt tttagcgttc ctttacttga
     7141 taatttatcc aaaaaccttc attcgtcaaa acaattatga tataattaat ttatgatatg
     7201 attaattaat atttacggat aactaatcat ttcaatgcaa tagatcaaca tgcaaatcaa
     7261 tatccacctt actccgaaca ttttgcacca acgaatgccc agcactatat cccggcaagc
     7321 aatataacgt ctataatttt ttatgataac gataaaatta aattttatga aaggggtaaa
     7381 agcatgagaa aaatagctgt attgttaatt actttgacgt tcttagtcac gactcgaatt
     7441 taggaggctt acttgtctgc tttcttcatt agaatcaatc cttttttaaa agtcaatccc
     7501 gtttgttgaa ctactcttta ataaaataat ttttccgttc ccaattccac attgcaataa
     7561 tagaaaatcc atcttcatcg gctttttcgt catcatctgt atgaatcaaa tcgccttctt
     7621 ctgtgtcatc aaggtttaat tttttatgta tttcttttaa caaaccacca taggagatta
     7681 accttttacg gtgtaaacct tcctccaaat cagacaaacg tttcaaattc ttttcttcat
     7741 catcggtcat aaaatccgta tcctttacag gatattttgc agtttcgtca attgccgatt
     7801 gtatatccga tttatattta tttttcggtc gaatcatttg aacttttaca tttggatcat
     7861 agtctaattt cattgccttt ttccaaaatt gaatccattg tttttgattc acgtagtttt
     7921 ctgtattctt aaaataagtt ggttccacac ataccaatac atgcatgtgc tgattataag
     7981 aattatcttt attatttatt gtcacttccg ttgcacgcat aaaaccaaca agatttttat
     8041 taattttttt atattgcatc attcggcgaa atccttgagc catatctgac aaactcttat
     8101 ttaattcttc gccatcataa acatttttaa ctgttaatgt gagaaacaac caacgaactg
     8161 ttggcttttg tttaataact tcagcaacaa ccttttgtga ctgaatgcca tgtttcattg
     8221 ctctcctcca gttgcacatt ggacaaagcc tggatttaca aaaccacact cgatacaact
     8281 ttctttcgcc tgtttcacga ttttgtttat actctaatat ttcagcacaa tcttttactc
     8341 tttcagcctt tttaaattca agaatatgca gaagttcaaa gtaatcaaca ttagcgattt
     8401 tcttttctct ccatggtctc acttttccac tttttgtctt gtccactaaa acccttgatt
     8461 tttcatctga ataaatgcta ctattaggac acataatatt aaaagaaacc cccat
//
