LOCUS       V005308                 8544 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V005308
VERSION     V005308
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8544)
  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 8544)
  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 8544)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8544)
  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..8544
                     /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="APQ46108.1"
                     /translation="MENLSKDIDEILITEEELKEKIKELGRQITKDYKGKNLMLVGVLK
                     GALMFMADLSRHIDLPLSLDFMAVSSYGSSTHSSGIVKIIKDLDISIEGKDVLIVEDII
                     DSGLTLSYLRETLLGRKPKSLKICTILDKPERREASVKVDYVGFKIPDKFVVGYGLDFD
                     EKYRNLPFIGVLKPEMYS"
     terminator      5142..5330
                     /label="CYC1 terminator"
                     /note="transcription terminator for CYC1"
     rep_origin      complement(5590..6178)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6265..6843)
                     /codon_start=1
                     /product="Tdk"
                     /EC_number="2.7.1.21"
                     /label="Tdk"
                     /note="thymidine kinase"
                     /protein_id="APQ46109.1"
                     /translation="MYGPKDHGYIEVVTGPMFSGKSEELIRRIKRAKIARQKVQVFKPA
                     IDDRYSIDKVVSHNGDNMHAIAIVKASDILAYAEEDTDVFAIDEVQFFDSEIVDIVKEI
                     ADSGKRVICAGLDMDFRGEPFGPTPELMAIAEFVDKLTAICMKCGNPATRTQRLINGKP
                     ANYDDPIIMVGAKESYEARCRKCHEVPRT"
     CDS             complement(7543..8544)
                     /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 ataattcctc
     1621 tcccatcttt tttatttata tagagatatt tgcatataag caaattactc cccgcaataa
     1681 tgctagttac caatatacac ctttgtactg tatatggtat acgccattgt ataccctcaa
     1741 ataaatgttt gaggaattaa caaaatgaca ttttgaatta tagcaaaacc gtaaccatat
     1801 tgcaagagtt ggaacaaata atttcgaaat ttaatcttgc ccttgctttt atcttttcgt
     1861 tcaaagtgtg tatttttctt tgattaaaca atttaacaat tttttcattc gtttttttat
     1921 tctcaatttc tgcaagaaac cattcggcaa actttcattt tcttttgttt accatatttt
     1981 tatccgtata tatcccgtat ttgtaacata ataccatata gttaatacaa tccctataat
     2041 ttcgattgac ttaaagaaca tcaaaagtta aaattagaat aaccttacga ttgagaaaag
     2101 ggaggtccta agctagtttt attgaattga atgatataaa attggctaaa aaattcccta
     2161 taaaattcat gacatccatg gtacaaatat aaaatcgtaa caaaaaataa taatgagatt
     2221 gtaaaatatt ttatgtaaac taattgactc cttctatgat agaataataa agtataggag
     2281 gagtttttta tggcaagaaa aaggataata acaccagaaa agaaagagct tatcagaaat
     2341 ctcatttctg agtacaacat tacttcagca aaggatttgc aggaagcatt gaaggatctg
     2401 ctcggagata cgatacaaaa tatgttggaa gcagagctgg atgaacatct cggatatgaa
     2461 aagtacgaat caactgaaga agcgaaatca aattaccgta acgggtacac atcaaaaaca
     2521 ttaaagtcaa gtgtagggca agtggaaata gatatcccgc gggaccggaa tgcagaattc
     2581 gagccgaaaa ttgttcccag gtataaaagg gacaatcgtg 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 accacttctt atgaaaagcc tgaaaaaaac
     4621 ggcaaaaggg tggcagtaat aggttcggga cctgcaagcc ttacctgtgc aagcgacctg
     4681 gccaaactcg gctacgaagt aacaatcttc gaagcctttc acgtgcccgg cggagtgttg
     4741 atgtacggta ttccggaatt caggcttcca aagaaactgg ttcaggagga aattgaaacc
     4801 ataaagcagc tgggtgtgga aattaaaaca aatatggtta taggaaaggt ttattccatt
     4861 gacgaactca aagctgaagg atatgatgcc atatttatag gctcgggtgc cggattgcct
     4921 tcatttatga aaattcccgg agaaaacctc aacggagttt actcggcaaa tgagtttctc
     4981 acaagaataa acctcatgaa ggcttatgaa ttccccaact gcgatactcc cgtgaaagta
     5041 ggaaagaatg tcgccgttgt gggcggagga aatgtcgcaa tggacgccgc aagaagcgca
     5101 aaaagacttc tgctaatagt agtgaaaaaa tcagggataa tgccgctcta accgaaaagg
     5161 aaggagttag acaacctgaa gtctaggtcc ctatttattt ttttatagtt atgttagtat
     5221 taagaacgtt atttatattt caaatttttc ttttttttct gtacagacgc gtgtacgcat
     5281 gtaacattat actgaaaacc ttgcttgaga aggttttggg acgctcgaag gctttaattt
     5341 gcaagctgcg gccctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5401 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5461 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5521 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5581 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5641 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5701 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5761 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5821 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5881 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5941 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6001 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6061 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6121 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6181 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6241 gattttggtc atgaaaaaat ccgcttaagt ccgcgggact tcatggcact ttctacacct
     6301 tgcttcataa gactcctttg ctccaaccat tataatggga tcgtcgtaat tggcaggctt
     6361 cccatttata agcctttgcg tgcgagtagc aggattgcca cacttcatgc atatagcagt
     6421 aagcttatcg acaaattcag ctatggccat caattctgga gtcggaccaa atggttcacc
     6481 tctaaagtcc atgtcaagcc ctgcacatat gactcttttc ccgctatcgg caatctcttt
     6541 tacgatgtcg actatttcag aatcaaaaaa ttgaacttca tctatagcaa atacatccgt
     6601 atcttcttca gcataagcca atatgtcaga agcctttact atggcaatgg cgtgcatgtt
     6661 gtcgccgtta tgagatacga ctttgtctat ggaataccta tcgtctatag ccggtttgaa
     6721 aacctgaact ttttgcctgg caatcttagc tctctttatc cttcttataa gttcctcact
     6781 ttttccactg aacatgggac cagttacaac ttctatgtac ccgtggtctt taggcccata
     6841 catctttcgt cctccttaaa attttcgttt atttgttttt tatttagtgt gcctaaaagc
     6901 accatcttat acattcgtac cttatatctc ttatcttgta tctcttacct catatctctt
     6961 attattaact accgaagtat attttcataa cggaatctta tgacttattt ttctgactgc
     7021 cgttttacca tacggttaat ttatacggaa acagctaatc taaccttgct gaaggcatgt
     7081 ttaccaaaat attatatcta ctttaccata ttaattcaag tggtcatttt gatattttac
     7141 gccaaaattt ttagcgttcc tttacttgat aatttatcca aaaaccttca ttcgtcaaaa
     7201 caattatgat ataattaatt tatgatatga ttaattaata tttacggata actaatcatt
     7261 tcaatgcaat agatcaacat gcaaatcaat atccacctta ctccgaacat tttgcaccaa
     7321 cgaatgccca gcactatatc ccggcaagca atataacgtc tataattttt tatgataacg
     7381 ataaaattaa attttatgaa aggggtaaaa gcatgagaaa aatagctgta ttgttaatta
     7441 ctttgacgtt cttagtcacg actcgaattt aggaggctta cttgtctgct ttcttcatta
     7501 gaatcaatcc ttttttaaaa gtcaatcccg tttgttgaac tactctttaa taaaataatt
     7561 tttccgttcc caattccaca ttgcaataat agaaaatcca tcttcatcgg ctttttcgtc
     7621 atcatctgta tgaatcaaat cgccttcttc tgtgtcatca aggtttaatt ttttatgtat
     7681 ttcttttaac aaaccaccat aggagattaa ccttttacgg tgtaaacctt cctccaaatc
     7741 agacaaacgt ttcaaattct tttcttcatc atcggtcata aaatccgtat cctttacagg
     7801 atattttgca gtttcgtcaa ttgccgattg tatatccgat ttatatttat ttttcggtcg
     7861 aatcatttga acttttacat ttggatcata gtctaatttc attgcctttt tccaaaattg
     7921 aatccattgt ttttgattca cgtagttttc tgtattctta aaataagttg gttccacaca
     7981 taccaataca tgcatgtgct gattataaga attatcttta ttatttattg tcacttccgt
     8041 tgcacgcata aaaccaacaa gatttttatt aattttttta tattgcatca ttcggcgaaa
     8101 tccttgagcc atatctgaca aactcttatt taattcttcg ccatcataaa catttttaac
     8161 tgttaatgtg agaaacaacc aacgaactgt tggcttttgt ttaataactt cagcaacaac
     8221 cttttgtgac tgaatgccat gtttcattgc tctcctccag ttgcacattg gacaaagcct
     8281 ggatttacaa aaccacactc gatacaactt tctttcgcct gtttcacgat tttgtttata
     8341 ctctaatatt tcagcacaat cttttactct ttcagccttt ttaaattcaa gaatatgcag
     8401 aagttcaaag taatcaacat tagcgatttt cttttctctc catggtctca cttttccact
     8461 ttttgtcttg tccactaaaa cccttgattt ttcatctgaa taaatgctac tattaggaca
     8521 cataatatta aaagaaaccc ccat
//
