LOCUS       V005305                 8513 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V005305
VERSION     V005305
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8513)
  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 8513)
  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 8513)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8513)
  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..8513
                     /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             3191..3838
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     CDS             3859..4404
                     /codon_start=1
                     /product="Hpt"
                     /EC_number="2.4.2.8"
                     /label="Hpt"
                     /note="hypoxanthine phosphoribosyltransferase"
                     /protein_id="APQ46118.1"
                     /translation="MENLSKDIDEILITEEELKEKIKELGRQITKDYKGKNLMLVGVLK
                     GALMFMADLSRHIDLPLSLDFMAVSSYGSSTHSSGIVKIIKDLDISIEGKDVLIVEDII
                     DSGLTLSYLRETLLGRKPKSLKICTILDKPERREASVKVDYVGFKIPDKFVVGYGLDFD
                     EKYRNLPFIGVLKPEMYS"
     terminator      5111..5299
                     /label="CYC1 terminator"
                     /note="transcription terminator for CYC1"
     rep_origin      complement(5559..6147)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6234..6812)
                     /codon_start=1
                     /product="Tdk"
                     /EC_number="2.7.1.21"
                     /label="Tdk"
                     /note="thymidine kinase"
                     /protein_id="APQ46119.1"
                     /translation="MYGPKDHGYIEVVTGPMFSGKSEELIRRIKRAKIARQKVQVFKPA
                     IDDRYSIDKVVSHNGDNMHAIAIVKASDILAYAEEDTDVFAIDEVQFFDSEIVDIVKEI
                     ADSGKRVICAGLDMDFRGEPFGPTPELMAIAEFVDKLTAICMKCGNPATRTQRLINGKP
                     ANYDDPIIMVGAKESYEARCRKCHEVPRT"
     CDS             complement(7512..8513)
                     /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 ataaaactga
     1621 tacgaatgaa tatcgtgatt acagttgttt attgaaagac agcataaatt agttgctaaa
     1681 aattggttac taaaagatat tttaatttgg caaattctta tttgacttat ttacaagttg
     1741 caaatataat attcagtgga aatatattta gaggaagaac atattcagaa aaaagcagcc
     1801 tatcgaaaaa acaaaaaagc ctgtcaatgg ggatgggttt tatcactcta gtcgaacagc
     1861 ttgctacaga aattgctcat ttttagttgg tgtttcgtga tattctatgt tgtatacaat
     1921 ataccgccag aaatcacaga aatccagtaa ttttgcatgg tagtaacaat tgcctgattt
     1981 gtacaaatca aaatatttta ataaaattgc cgatgtgcta ttataaaaaa gtgagtaatt
     2041 aaaataggca tatgggtggt atataaaatt ttactgtgga atataaaaat tatttaaaat
     2101 ggggttatac gaatgttaag tgtgaaataa agtttggttc ttttgacata aagctcagcg
     2161 gatgctgagc ttaatatatt tgtaattttt agatttgggt tattatctgt gtggcttttt
     2221 tgtaaccagt atttggtttc tgccgttttt aaatgatttt ctaacatcat agccccttgt
     2281 taccaatgtg tcgtatattt tgtcggtttg ctcaaagtag ttgttttcca catccaaaat
     2341 cagtgtttct ccgggcttta actcgtttga caacttttcc agtgtggcat ccacgttcat
     2401 attttctgcg tttatgattt tctcaccgtt gtgattcatt ttgatttcct cccgttttgg
     2461 aaaaaataaa tgaaaaatga tttaagtcag tgcaaaaaat gtttcaaaac tcaattcaat
     2521 gcgtatatat tttgtatata gtattgacat gcagatgcta ttatataagg caaatattat
     2581 tatattgtaa aaaatttctt gtttgaggag caatcgtggg aataggcatg gaaaagattt
     2641 ttttgcccga ggatgcattg ataaagtatt tcttttccga aagagaggaa aagattctaa
     2701 agagttttgg aaatactgat gaatattgtg tgcagagtac aattctatgg acaagaaaag
     2761 aggctttgtc aaaacttttt cgtctgggaa tgaggatgga ttttaaaaag ctggatactt
     2821 tggagggcga ggtggttttt caggaaacaa acagggcgcg tctgttttct tttatatgca
     2881 ataattactg tatctctctg gcattgccag gttttaataa agattaaaat tattgactag
     2941 aaataaaaaa attgtccata atattaatgg acaaaaaaac aaagaattac atcaaaggaa
     3001 gataaaaata ctttgttaaa aaattaatta ttttttatct aaactattga aaatgaaaat
     3061 aaaataatat aaaatgaatc atagtgcaag agatacttgc cagaggatga atattttact
     3121 gcattcatgc tttatggcag ctaatagagg cattaaatta aattttaatt tacaatagga
     3181 ggcgatatta atgaacttta ataaaattga tttagacaat tggaagagaa aagagatatt
     3241 taatcattat ttgaaccaac aaacgacttt tagtataacc acagaaattg atattagtgt
     3301 tttataccga aacataaaac aagaaggata taaattttac cctgcattta ttttcttagt
     3361 gacaagggtg ataaactcaa atacagcttt tagaactggt tacaatagcg acggagagtt
     3421 aggttattgg gataagttag agccacttta tacaattttt gatggtgtat ctaaaacatt
     3481 ctctggtatt tggactcctg taaagaatga cttcaaagag ttttatgatt tatacctttc
     3541 tgatgtagag aaatataatg gttcggggaa attgtttccc aaaacaccta tacctgaaaa
     3601 tgctttttct ctttctatta ttccatggac ttcatttact gggtttaact taaatatcaa
     3661 taataatagt aattaccttc tacccattat tacagcagga aaattcatta ataaaggtaa
     3721 ttcaatatat ttaccgctat ctttacaggt acatcattct gtttgtgatg gttatcatgc
     3781 aggattgttt atgaactcta ttcaggaatt gtcagatagg cctaatgact ggcttttata
     3841 ataaaggagg tcgacgtcat ggaaaattta tcaaaagaca tcgatgaaat tttgatcaca
     3901 gaagaagaac ttaaggaaaa gataaaagag cttgggaggc aaatcacaaa agactacaaa
     3961 gggaaaaatt tgatgttggt aggagtttta aaaggtgctt taatgtttat ggctgatttg
     4021 tcaagacaca tagatttgcc tttatcactt gattttatgg ctgtttccag ctatggaagc
     4081 tcaactcatt catcaggaat agtaaagata atcaaagatc ttgatataag catagaaggc
     4141 aaagatgttc tgattgtgga agacataatt gacagcggtt tgactttgtc ttacttaagg
     4201 gaaactttac ttggaaggaa gccaaaaagc ctgaaaatat gcacaatatt agacaaaccg
     4261 gagagaagag aagcatctgt aaaagtcgat tatgtaggat ttaagatacc tgataagttt
     4321 gtcgtgggtt atggattgga ctttgatgaa aagtacagga accttccttt tataggcgtt
     4381 ttgaaacctg aaatgtacag ctaagcccgg gcctcgagaa aacaaaaggc tcagtcggaa
     4441 gactgggcct tttgttttgg taccgaattc ggcgcgcctc agcgtttaaa cccgctgatc
     4501 ctagagggcc gcatcatgta attagttatg tcacgcttac attcacgccc tccccatgct
     4561 ttaatacata cctggcccag tagttcagat gagtggtttt aagactgtaa cgaaagggat
     4621 aataaaggaa aacccggtat tcaggcttgt ccttggtaca tgccccactt tggctgtgac
     4681 aacatcagcc gaaaatgcct tgggaatggg ggcggccgtt atatttgtgt tgatggggtc
     4741 aaatgtagtg atttcccttt tgagaaaggt gatacctgac aaggtcagaa ttcctgcata
     4801 tataacaatt atttcgggat ttgtgacgat agttcagatg ctgattaagg catatcttcc
     4861 tgctattgat aagcagctgg gtatatttat acctcttata gttgtaaact gtatcatact
     4921 tgcaagagcg gagatgtttg caagcaagaa cggtccactc atgtcaattc ttgacgggct
     4981 tggcatggga ataggtttta ccttgtcgct tcttgttatg ggaagcgtca gagagatact
     5041 tggaaacggc acatggtttg gaatgaatat taccggtgct gctaatagta gtgaaaaaat
     5101 cagggataat gccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc
     5161 tatttatttt tttatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct
     5221 tttttttctg tacagacgcg tgtacgcatg taacattata ctgaaaacct tgcttgagaa
     5281 ggttttggga cgctcgaagg ctttaatttg caagctgcgg ccctgcatta atgaatcggc
     5341 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     5401 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5461 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5521 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5581 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5641 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5701 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5761 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5821 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5881 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5941 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     6001 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6061 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6121 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6181 gctcagtgga acgaaaactc acgttaaggg attttggtca tgaaaaaatc cgcttaagtc
     6241 cgcgggactt catggcactt tctacacctt gcttcataag actcctttgc tccaaccatt
     6301 ataatgggat cgtcgtaatt ggcaggcttc ccatttataa gcctttgcgt gcgagtagca
     6361 ggattgccac acttcatgca tatagcagta agcttatcga caaattcagc tatggccatc
     6421 aattctggag tcggaccaaa tggttcacct ctaaagtcca tgtcaagccc tgcacatatg
     6481 actcttttcc cgctatcggc aatctctttt acgatgtcga ctatttcaga atcaaaaaat
     6541 tgaacttcat ctatagcaaa tacatccgta tcttcttcag cataagccaa tatgtcagaa
     6601 gcctttacta tggcaatggc gtgcatgttg tcgccgttat gagatacgac tttgtctatg
     6661 gaatacctat cgtctatagc cggtttgaaa acctgaactt tttgcctggc aatcttagct
     6721 ctctttatcc ttcttataag ttcctcactt tttccactga acatgggacc agttacaact
     6781 tctatgtacc cgtggtcttt aggcccatac atctttcgtc ctccttaaaa ttttcgttta
     6841 tttgtttttt atttagtgtg cctaaaagca ccatcttata cattcgtacc ttatatctct
     6901 tatcttgtat ctcttacctc atatctctta ttattaacta ccgaagtata ttttcataac
     6961 ggaatcttat gacttatttt tctgactgcc gttttaccat acggttaatt tatacggaaa
     7021 cagctaatct aaccttgctg aaggcatgtt taccaaaata ttatatctac tttaccatat
     7081 taattcaagt ggtcattttg atattttacg ccaaaatttt tagcgttcct ttacttgata
     7141 atttatccaa aaaccttcat tcgtcaaaac aattatgata taattaattt atgatatgat
     7201 taattaatat ttacggataa ctaatcattt caatgcaata gatcaacatg caaatcaata
     7261 tccaccttac tccgaacatt ttgcaccaac gaatgcccag cactatatcc cggcaagcaa
     7321 tataacgtct ataatttttt atgataacga taaaattaaa ttttatgaaa ggggtaaaag
     7381 catgagaaaa atagctgtat tgttaattac tttgacgttc ttagtcacga ctcgaattta
     7441 ggaggcttac ttgtctgctt tcttcattag aatcaatcct tttttaaaag tcaatcccgt
     7501 ttgttgaact actctttaat aaaataattt ttccgttccc aattccacat tgcaataata
     7561 gaaaatccat cttcatcggc tttttcgtca tcatctgtat gaatcaaatc gccttcttct
     7621 gtgtcatcaa ggtttaattt tttatgtatt tcttttaaca aaccaccata ggagattaac
     7681 cttttacggt gtaaaccttc ctccaaatca gacaaacgtt tcaaattctt ttcttcatca
     7741 tcggtcataa aatccgtatc ctttacagga tattttgcag tttcgtcaat tgccgattgt
     7801 atatccgatt tatatttatt tttcggtcga atcatttgaa cttttacatt tggatcatag
     7861 tctaatttca ttgccttttt ccaaaattga atccattgtt tttgattcac gtagttttct
     7921 gtattcttaa aataagttgg ttccacacat accaatacat gcatgtgctg attataagaa
     7981 ttatctttat tatttattgt cacttccgtt gcacgcataa aaccaacaag atttttatta
     8041 atttttttat attgcatcat tcggcgaaat ccttgagcca tatctgacaa actcttattt
     8101 aattcttcgc catcataaac atttttaact gttaatgtga gaaacaacca acgaactgtt
     8161 ggcttttgtt taataacttc agcaacaacc ttttgtgact gaatgccatg tttcattgct
     8221 ctcctccagt tgcacattgg acaaagcctg gatttacaaa accacactcg atacaacttt
     8281 ctttcgcctg tttcacgatt ttgtttatac tctaatattt cagcacaatc ttttactctt
     8341 tcagcctttt taaattcaag aatatgcaga agttcaaagt aatcaacatt agcgattttc
     8401 ttttctctcc atggtctcac ttttccactt tttgtcttgt ccactaaaac ccttgatttt
     8461 tcatctgaat aaatgctact attaggacac ataatattaa aagaaacccc cat
//
