LOCUS       V007997                 8705 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V007997
VERSION     V007997
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8705)
  AUTHORS   Hon S, Holwerda EK, Worthen RS, Maloney MI, Tian L, Cui J, Lin PP,
            Lynd LR, Olson DG.
  TITLE     Expressing the Thermoanaerobacterium saccharolyticum pforA in
            engineered Clostridium thermocellum improves ethanol production
  JOURNAL   Biotechnol Biofuels 11, 242 (2018)
   PUBMED   30202437
REFERENCE   2  (bases 1 to 8705)
  AUTHORS   Hon S, Olson DG, Holwerda EK, Worthen RS, Maloney MI, Tian L, Cui J,
            Lin PP, Lynd LR.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-APR-2018) Thayer School of Engineering, Dartmouth
            College, 14 Engineering Drive, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 8705)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8705)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "Biotechnol
            Biofuels 11, 242 (2018)"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (20-APR-2018) Thayer School of Engineering, Dartmouth College, 14
            Engineering Drive, Hanover, NH 03755, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8705
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(60..648)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(735..1313)
                     /codon_start=1
                     /gene="tdk"
                     /product="thymidine kinase"
                     /label="tdk"
                     /protein_id="AYA22189.1"
                     /translation="MYGPKDHGYIEVVTGPMFSGKSEELIRRIKRAKIARQKVQVFKPA
                     IDDRYSIDKVVSHNGDNMHAIAIVKASDILAYAEEDTDVFAIDEVQFFDSEIVDIVKEI
                     ADSGKRVICAGLDMDFRGEPFGPTPELMAIAEFVDKLTAICMKCGNPATRTQRLINGKP
                     ANYDDPIIMVGAKESYEARCRKCHEVPRT"
     gene            complement(735..1313)
                     /gene="tdk"
                     /label="tdk"
     regulatory      complement(1314..1934)
                     /label="C. thermocellum cbp promoter"
                     /note="C. thermocellum cbp promoter"
                     /regulatory_class="promoter"
     CDS             complement(2013..3014)
                     /label="repB"
                     /note="RepB replication protein"
     rep_origin      complement(3015..3469)
                     /direction=LEFT
                     /label="C. thermocellum origin of replication"
                     /note="C. thermocellum origin of replication"
     CDS             complement(3575..4432)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(4433..4537)
                     /label="AmpR promoter"
     misc_feature    4627..5171
                     /label="upstream homology recombination flank"
                     /note="upstream homology recombination flank"
     misc_feature    5172..5711
                     /label="downstream homology recombination flank"
                     /note="downstream homology recombination flank"
     regulatory      5715..6289
                     /label="C. thermocellum gapdh promoter"
                     /note="C. thermocellum gapdh promoter"
                     /regulatory_class="promoter"
     CDS             6290..6937
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     CDS             6958..7503
                     /codon_start=1
                     /gene="hpt"
                     /product="hypoxanthine phosphoribosyltransferase"
                     /label="hpt"
                     /protein_id="AYA22193.1"
                     /translation="MENLSKDIDEILITEEELKEKIKELGRQITKDYKGKNLMLVGVLK
                     GALMFMADLSRHIDLPLSLDFMAVSSYGSSTHSSGIVKIIKDLDISIEGKDVLIVEDII
                     DSGLTLSYLRETLLGRKPKSLKICTILDKPERREASVKVDYVGFKIPDKFVVGYGLDFD
                     EKYRNLPFIGVLKPEMYS"
     gene            6958..7503
                     /gene="hpt"
                     /label="hpt"
     misc_feature    7688..8284
                     /label="gene target internal homology recombination flank"
                     /note="gene target internal homology recombination flank"
     terminator      8317..8505
                     /label="CYC1 terminator"
                     /note="transcription terminator for CYC1"
ORIGIN
        1 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
       61 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
      121 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
      181 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
      241 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
      301 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
      361 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
      421 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
      481 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
      541 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
      601 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
      661 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
      721 atgaaaaaat ccgcttaagt ccgcgggact tcatggcact ttctacacct tgcttcataa
      781 gactcctttg ctccaaccat tataatggga tcgtcgtaat tggcaggctt cccatttata
      841 agcctttgcg tgcgagtagc aggattgcca cacttcatgc atatagcagt aagcttatcg
      901 acaaattcag ctatggccat caattctgga gtcggaccaa atggttcacc tctaaagtcc
      961 atgtcaagcc ctgcacatat gactcttttc ccgctatcgg caatctcttt tacgatgtcg
     1021 actatttcag aatcaaaaaa ttgaacttca tctatagcaa atacatccgt atcttcttca
     1081 gcataagcca atatgtcaga agcctttact atggcaatgg cgtgcatgtt gtcgccgtta
     1141 tgagatacga ctttgtctat ggaataccta tcgtctatag ccggtttgaa aacctgaact
     1201 ttttgcctgg caatcttagc tctctttatc cttcttataa gttcctcact ttttccactg
     1261 aacatgggac cagttacaac ttctatgtac ccgtggtctt taggcccata catctttcgt
     1321 cctccttaaa attttcgttt atttgttttt tatttagtgt gcctaaaagc accatcttat
     1381 acattcgtac cttatatctc ttatcttgta tctcttacct catatctctt attattaact
     1441 accgaagtat attttcataa cggaatctta tgacttattt ttctgactgc cgttttacca
     1501 tacggttaat ttatacggaa acagctaatc taaccttgct gaaggcatgt ttaccaaaat
     1561 attatatcta ctttaccata ttaattcaag tggtcatttt gatattttac gccaaaattt
     1621 ttagcgttcc tttacttgat aatttatcca aaaaccttca ttcgtcaaaa caattatgat
     1681 ataattaatt tatgatatga ttaattaata tttacggata actaatcatt tcaatgcaat
     1741 agatcaacat gcaaatcaat atccacctta ctccgaacat tttgcaccaa cgaatgccca
     1801 gcactatatc ccggcaagca atataacgtc tataattttt tatgataacg ataaaattaa
     1861 attttatgaa aggggtaaaa gcatgagaaa aatagctgta ttgttaatta ctttgacgtt
     1921 cttagtcacg actcgaattt aggaggctta cttgtctgct ttcttcatta gaatcaatcc
     1981 ttttttaaaa gtcaatcccg tttgttgaac tactctttaa taaaataatt tttccgttcc
     2041 caattccaca ttgcaataat agaaaatcca tcttcatcgg ctttttcgtc atcatctgta
     2101 tgaatcaaat cgccttcttc tgtgtcatca aggtttaatt ttttatgtat ttcttttaac
     2161 aaaccaccat aggagattaa ccttttacgg tgtaaacctt cctccaaatc agacaaacgt
     2221 ttcaaattct tttcttcatc atcggtcata aaatccgtat cctttacagg atattttgca
     2281 gtttcgtcaa ttgccgattg tatatccgat ttatatttat ttttcggtcg aatcatttga
     2341 acttttacat ttggatcata gtctaatttc attgcctttt tccaaaattg aatccattgt
     2401 ttttgattca cgtagttttc tgtattctta aaataagttg gttccacaca taccaataca
     2461 tgcatgtgct gattataaga attatcttta ttatttattg tcacttccgt tgcacgcata
     2521 aaaccaacaa gatttttatt aattttttta tattgcatca ttcggcgaaa tccttgagcc
     2581 atatctgaca aactcttatt taattcttcg ccatcataaa catttttaac tgttaatgtg
     2641 agaaacaacc aacgaactgt tggcttttgt ttaataactt cagcaacaac cttttgtgac
     2701 tgaatgccat gtttcattgc tctcctccag ttgcacattg gacaaagcct ggatttacaa
     2761 aaccacactc gatacaactt tctttcgcct gtttcacgat tttgtttata ctctaatatt
     2821 tcagcacaat cttttactct ttcagccttt ttaaattcaa gaatatgcag aagttcaaag
     2881 taatcaacat tagcgatttt cttttctctc catggtctca cttttccact ttttgtcttg
     2941 tccactaaaa cccttgattt ttcatctgaa taaatgctac tattaggaca cataatatta
     3001 aaagaaaccc ccatctattt agttatttgt ttggtcactt ataactttaa cagatggggt
     3061 ttttctgtgc aaccaatttt aagggttttc aatactttaa aacacataca taccaacact
     3121 tcaacgcacc tttcagcaac taaaataaaa atgacgttat ttctatatgt atcaagaata
     3181 gaaagaactc gtttttcgct acgctcaaaa cgcaaaaaaa gcactcattc gagtgctttt
     3241 tcttatcgct ccaaatcatg cgattttttc ctctttgctt ttctttgctc acgaagttct
     3301 cgatcacgct gcaaaacatc ttgaagcgaa aaagtattct tcttttcttc cgatcgctca
     3361 tgctgacgca cgaaaagccc tctaggcgca taggaacaac tcctaaatgc atgtgagggg
     3421 ttttctcgtc catgtgaaca gtcgcatacg caatattttg tttcccatag cggccgcgat
     3481 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3541 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3601 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3661 ctacgatacg ggagcgctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     3721 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3781 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     3841 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     3901 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3961 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     4021 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4081 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     4141 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     4201 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     4261 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4321 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4381 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     4441 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4501 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     4561 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     4621 ggcgatgggc agatacctgg ttgttccgat gaaaaatctc ttaaaacttg tatataatgg
     4681 ctttatcggg ggaattgcca ttctggttat aaattttgtg ggaggaaact tcagcttcca
     4741 aatcgcattg aatcctctga cggcatttat tgtaggtacg ctgggagtgc ccggagtact
     4801 tatgataaca attctaaagt atcttttcca ggtatgaaaa cagcttaaaa tcaaccaagt
     4861 accctgtttt tgattttaaa tcccttattc caaattccat ataaagtaaa ctgtcaatta
     4921 aaaattaaaa ttttcgcaaa cgcataagaa attgtataca aagaacattg agagagcaaa
     4981 tcagcataga tattaagcca ttttcaattt cgctttcttg acaaaaaatt gacaaacaat
     5041 tatacttgaa gatgtattaa tctatgtttt tggtatacaa agctgaattt tttttcagcg
     5101 taaaacttct cataaataaa ataaggagga ttaccacatg ggcaaggttg ttgaaatcag
     5161 atggcacggg caacgaatat atgctggaag aaattcagaa agaagtcgac ttaagatggg
     5221 agagactctt gaagctggcc ggagaggctt aaaaaacgct aaagtaaaat gataataaaa
     5281 agacctgggt tttggcctgg gtctttaaat ttgtaaaatc agtttatttt ttaaaatgag
     5341 aaaatattat tgaaaatagc atttaaaagt ctcattaaag tatcatttaa gaaggacttt
     5401 cggtactttg ggttggtata aaaagaaaaa acaggccgaa cttgccgaca aaacggcaaa
     5461 aagagtgacg ataaaagcca ccaaacctaa tgtttttttt aacatgaaaa attactcccc
     5521 ctctatccaa ttattcattt aagattattg aagccggtat tgcggtcaga attattgcag
     5581 aaggaatgtg gttcgtagga tataggataa tatggttcat agaatgatac agtttcgtgt
     5641 aaacaaatca gtgttcaatg atatgcgatt ttaagcttat tagagcgttt ttggcttcat
     5701 tgattttttg tatcgtggga ataggcatgg aaaagatttt tttgcccgag gatgcattga
     5761 taaagtattt cttttccgaa agagaggaaa agattctaaa gagttttgga aatactgatg
     5821 aatattgtgt gcagagtaca attctatgga caagaaaaga ggctttgtca aaactttttc
     5881 gtctgggaat gaggatggat tttaaaaagc tggatacttt ggagggcgag gtggtttttc
     5941 aggaaacaaa cagggcgcgt ctgttttctt ttatatgcaa taattactgt atctctctgg
     6001 cattgccagg ttttaataaa gattaaaatt attgactaga aataaaaaaa ttgtccataa
     6061 tattaatgga caaaaaaaca aagaattaca tcaaaggaag ataaaaatac tttgttaaaa
     6121 aattaattat tttttatcta aactattgaa aatgaaaata aaataatata aaatgaatca
     6181 tagtgcaaga gatacttgcc agaggatgaa tattttactg cattcatgct ttatggcagc
     6241 taatagaggc attaaattaa attttaattt acaataggag gcgatattaa tgaactttaa
     6301 taaaattgat ttagacaatt ggaagagaaa agagatattt aatcattatt tgaaccaaca
     6361 aacgactttt agtataacca cagaaattga tattagtgtt ttataccgaa acataaaaca
     6421 agaaggatat aaattttacc ctgcatttat tttcttagtg acaagggtga taaactcaaa
     6481 tacagctttt agaactggtt acaatagcga cggagagtta ggttattggg ataagttaga
     6541 gccactttat acaatttttg atggtgtatc taaaacattc tctggtattt ggactcctgt
     6601 aaagaatgac ttcaaagagt tttatgattt atacctttct gatgtagaga aatataatgg
     6661 ttcggggaaa ttgtttccca aaacacctat acctgaaaat gctttttctc tttctattat
     6721 tccatggact tcatttactg ggtttaactt aaatatcaat aataatagta attaccttct
     6781 acccattatt acagcaggaa aattcattaa taaaggtaat tcaatatatt taccgctatc
     6841 tttacaggta catcattctg tttgtgatgg ttatcatgca ggattgttta tgaactctat
     6901 tcaggaattg tcagataggc ctaatgactg gcttttataa taaaggaggt cgacgtcatg
     6961 gaaaatttat caaaagacat cgatgaaatt ttgatcacag aagaagaact taaggaaaag
     7021 ataaaagagc ttgggaggca aatcacaaaa gactacaaag ggaaaaattt gatgttggta
     7081 ggagttttaa aaggtgcttt aatgtttatg gctgatttgt caagacacat agatttgcct
     7141 ttatcacttg attttatggc tgtttccagc tatggaagct caactcattc atcaggaata
     7201 gtaaagataa tcaaagatct tgatataagc atagaaggca aagatgttct gattgtggaa
     7261 gacataattg acagcggttt gactttgtct tacttaaggg aaactttact tggaaggaag
     7321 ccaaaaagcc tgaaaatatg cacaatatta gacaaaccgg agagaagaga agcatctgta
     7381 aaagtcgatt atgtaggatt taagatacct gataagtttg tcgtgggtta tggattggac
     7441 tttgatgaaa agtacaggaa ccttcctttt ataggcgttt tgaaacctga aatgtacagc
     7501 taagcccggg cctcgagaaa acaaaaggct cagtcggaag actgggcctt ttgttttggt
     7561 accgaattcg gcgcgcctca gcgtttaaac ccgctgatcc tagagggccg catcatgtaa
     7621 ttagttatgt cacgcttaca ttcacgccct ccccatgctt taatacatac ctggcccagt
     7681 agttcagcaa cactccaaaa actcctgacg aaataagacc tcttttaaaa ggctacaagg
     7741 gaaaggtttg cacaattgac gcaagaaaaa tttccattga aactttggga aaatacttcc
     7801 cgaacacccc aatgcttggt gctgttgtaa aggttagcaa gataatggac gaagaagaat
     7861 tcctcaagga tatggttgag tcattcaagc acaagtttgc gaacaagcct gaagttgttg
     7921 aaggcaatat caaggccctg gagaggtcaa tgcaggaggt gaaaggatta tgagtaaaga
     7981 attgagagat gtaaaacctg atgttacctg gaaagagata acatccggtg gagtgataga
     8041 ttcccctgga aacgcacatc tttttaagac cggtgactgg cgttctatga agccggtgtg
     8101 gaatgaggaa aaatgcaagc agtgtttgct ttgcaacccg gtatgtcctg attcatccat
     8161 aatggtaagt gaagagggaa aaatgacagg tatcgattat gatcactgca aaggatgtgg
     8221 catttgttca aaggtttgtc ctttcaaagc catagacttt gtagaggaag tataagagag
     8281 ggggctgcta atagtagtga aaaaatcagg gataatgccg ctctaaccga aaaggaagga
     8341 gttagacaac ctgaagtcta ggtccctatt tattttttta tagttatgtt agtattaaga
     8401 acgttattta tatttcaaat ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac
     8461 attatactga aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt aatttgcaag
     8521 ctgcggccct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     8581 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     8641 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     8701 agaac
//
