LOCUS       V009509                 8662 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V009509
VERSION     V009509
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8662)
  AUTHORS   Olson DG.
  TITLE     Transformation of Clostridium thermocellum by electroporation
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8662)
  AUTHORS   Olson DG.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-OCT-2011) Thayer School of Engineering, Dartmouth
            College, 8000 Cummings, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 8662)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8662)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (19-OCT-2011) Thayer School of Engineering, Dartmouth College, 8000
            Cummings, Hanover, NH 03755, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8662
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    11..514
                     /label="CEN/ARS"
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     regulatory      692..1339
                     /gene="GapDH"
                     /regulatory_class="promoter"
     gene            692..1339
                     /gene="GapDH"
                     /label="GapDH"
     CDS             1340..1987
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     CDS             2008..2553
                     /codon_start=1
                     /gene="hpt"
                     /product="hypoxanthine phosphoribosyltransferase"
                     /label="hpt"
                     /protein_id="AFC37773.1"
                     /translation="MENLSKDIDEILITEEELKEKIKELGRQITKDYKGKNLMLVGVLK
                     GALMFMADLSRHIDLPLSLDFMAVSSYGSSTHSSGIVKIIKDLDISIEGKDVLIVEDII
                     DSGLTLSYLRETLLGRKPKSLKICTILDKPERREASVKVDYVGFKIPDKFVVGYGLDFD
                     EKYRNLPFIGVLKPEMYS"
     gene            2008..2553
                     /gene="hpt"
                     /label="hpt"
     terminator      2663..2910
                     /label="CYC1 terminator"
                     /note="transcription terminator for CYC1"
     rep_origin      complement(3158..3746)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(3833..4411)
                     /codon_start=1
                     /gene="tdk"
                     /product="thymidine kinase"
                     /label="tdk"
                     /protein_id="AFC37774.1"
                     /translation="MYGPKDHGYIEVVTGPMFSGKSEELIRRIKRAKIARQKVQVFKPA
                     IDDRYSIDKVVSHNGDNMHAIAIVKASDILAYAEEDTDVFAIDEVQFFDSEIVDIVKEI
                     ADSGKRVICAGLDMDFRGEPFGPTPELMAIAEFVDKLTAICMKCGNPATRTQRLINGKP
                     ANYDDPIIMVGAKESYEARCRKCHEVPRT"
     gene            complement(3833..4411)
                     /gene="tdk"
                     /label="tdk"
     regulatory      complement(4412..5032)
                     /gene="Cbp"
                     /regulatory_class="promoter"
     gene            complement(4412..5032)
                     /gene="Cbp"
                     /label="Cbp"
     CDS             complement(5111..6112)
                     /label="repB"
                     /note="RepB replication protein"
     rep_origin      complement(6113..6567)
                     /direction=LEFT
                     /label="cth ori"
                     /note="cth ori"
     CDS             complement(6673..7530)
                     /label="AmpR"
                     /note="beta-lactamase"
     CDS             complement(7629..8429)
                     /label="URA3"
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     promoter        complement(8430..8645)
                     /label="URA3 promoter"
ORIGIN
        1 ggtccttttc atcacgtgct ataaaaataa ttataattta aattttttaa tataaatata
       61 taaattaaaa atagaaagta aaaaaagaaa ttaaagaaaa aatagttttt gttttccgaa
      121 gatgtaaaag actctagggg gatcgccaac aaatactacc ttttatcttg ctcttcctgc
      181 tctcaggtat taatgccgaa ttgtttcatc ttgtctgtgt agaagaccac acacgaaaat
      241 cctgtgattt tacattttac ttatcgttaa tcgaatgtat atctatttaa tctgcttttc
      301 ttgtctaata aatatatatg taaagtacgc tttttgttga aattttttaa acctttgttt
      361 attttttttt cttcattccg taactcttct accttcttta tttactttct aaaatccaaa
      421 tacaaaacat aaaaataaat aaacacagag taaattccca aattattcca tcattaaaag
      481 atacgaggcg cgtgtaagtt acaggcaagc gatccgtccg ccggcgaacg tggcgagaaa
      541 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
      601 gcgcgtaacc accacacccg ccgcgcttaa tggggcgcta cagggcgcgt ggggatgatc
      661 cactagtaag cttggatcct cgcgaggccg gccagtattc tgacatgggt gtatcaataa
      721 cccatgcgtt tccgtattgt atcggaatgg tttcggacag ggcggtggga ataggcatgg
      781 aaaagatttt tttgcccgag gatgcattga taaagtattt cttttccgaa agagaggaaa
      841 agattctaaa gagttttgga aatactgatg aatattgtgt gcagagtaca attctatgga
      901 caagaaaaga ggctttgtca aaactttttc gtctgggaat gaggatggat tttaaaaagc
      961 tggatacttt ggagggcgag gtggtttttc aggaaacaaa cagggcgcgt ctgttttctt
     1021 ttatatgcaa taattactgt atctctctgg cattgccagg ttttaataaa gattaaaatt
     1081 attgactaga aataaaaaaa ttgtccataa tattaatgga caaaaaaaca aagaattaca
     1141 tcaaaggaag ataaaaatac tttgttaaaa aattaattat tttttatcta aactattgaa
     1201 aatgaaaata aaataatata aaatgaatca tagtgcaaga gatacttgcc agaggatgaa
     1261 tattttactg cattcatgct ttatggcagc taatagaggc attaaattaa attttaattt
     1321 acaataggag gcgatattaa tgaactttaa taaaattgat ttagacaatt ggaagagaaa
     1381 agagatattt aatcattatt tgaaccaaca aacgactttt agtataacca cagaaattga
     1441 tattagtgtt ttataccgaa acataaaaca agaaggatat aaattttacc ctgcatttat
     1501 tttcttagtg acaagggtga taaactcaaa tacagctttt agaactggtt acaatagcga
     1561 cggagagtta ggttattggg ataagttaga gccactttat acaatttttg atggtgtatc
     1621 taaaacattc tctggtattt ggactcctgt aaagaatgac ttcaaagagt tttatgattt
     1681 atacctttct gatgtagaga aatataatgg ttcggggaaa ttgtttccca aaacacctat
     1741 acctgaaaat gctttttctc tttctattat tccatggact tcatttactg ggtttaactt
     1801 aaatatcaat aataatagta attaccttct acccattatt acagcaggaa aattcattaa
     1861 taaaggtaat tcaatatatt taccgctatc tttacaggta catcattctg tttgtgatgg
     1921 ttatcatgca ggattgttta tgaactctat tcaggaattg tcagataggc ctaatgactg
     1981 gcttttataa taaaggaggt cgacgtcatg gaaaatttat caaaagacat cgatgaaatt
     2041 ttgatcacag aagaagaact taaggaaaag ataaaagagc ttgggaggca aatcacaaaa
     2101 gactacaaag ggaaaaattt gatgttggta ggagttttaa aaggtgcttt aatgtttatg
     2161 gctgatttgt caagacacat agatttgcct ttatcacttg attttatggc tgtttccagc
     2221 tatggaagct caactcattc atcaggaata gtaaagataa tcaaagatct tgatataagc
     2281 atagaaggca aagatgttct gattgtggaa gacataattg acagcggttt gactttgtct
     2341 tacttaaggg aaactttact tggaaggaag ccaaaaagcc tgaaaatatg cacaatatta
     2401 gacaaaccgg agagaagaga agcatctgta aaagtcgatt atgtaggatt taagatacct
     2461 gataagtttg tcgtgggtta tggattggac tttgatgaaa agtacaggaa ccttcctttt
     2521 ataggcgttt tgaaacctga aatgtacagc taagcccggg cctcgagaaa acaaaaggct
     2581 cagtcggaag actgggcctt ttgttttggt accgaattcg gcgcgcctca gcgtttaaac
     2641 ccgctgatcc tagagggccg catcatgtaa ttagttatgt cacgcttaca ttcacgccct
     2701 ccccccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct
     2761 atttattttt ttatagttat gttagtatta agaacgttat ttatatttca aatttttctt
     2821 ttttttctgt acagacgcgt gtacgcatgt aacattatac tgaaaacctt gcttgagaag
     2881 gttttgggac gctcgaaggc tttaatttgc aagctgcggc cctgcattaa tgaatcggcc
     2941 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     3001 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3061 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3121 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3181 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3241 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3301 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3361 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3421 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3481 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3541 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     3601 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3661 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3721 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3781 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gaaaaaatcc gcttaagtcc
     3841 gcgggacttc atggcacttt ctacaccttg cttcataaga ctcctttgct ccaaccatta
     3901 taatgggatc gtcgtaattg gcaggcttcc catttataag cctttgcgtg cgagtagcag
     3961 gattgccaca cttcatgcat atagcagtaa gcttatcgac aaattcagct atggccatca
     4021 attctggagt cggaccaaat ggttcacctc taaagtccat gtcaagccct gcacatatga
     4081 ctcttttccc gctatcggca atctctttta cgatgtcgac tatttcagaa tcaaaaaatt
     4141 gaacttcatc tatagcaaat acatccgtat cttcttcagc ataagccaat atgtcagaag
     4201 cctttactat ggcaatggcg tgcatgttgt cgccgttatg agatacgact ttgtctatgg
     4261 aatacctatc gtctatagcc ggtttgaaaa cctgaacttt ttgcctggca atcttagctc
     4321 tctttatcct tcttataagt tcctcacttt ttccactgaa catgggacca gttacaactt
     4381 ctatgtaccc gtggtcttta ggcccataca tctttcgtcc tccttaaaat tttcgtttat
     4441 ttgtttttta tttagtgtgc ctaaaagcac catcttatac attcgtacct tatatctctt
     4501 atcttgtatc tcttacctca tatctcttat tattaactac cgaagtatat tttcataacg
     4561 gaatcttatg acttattttt ctgactgccg ttttaccata cggttaattt atacggaaac
     4621 agctaatcta accttgctga aggcatgttt accaaaatat tatatctact ttaccatatt
     4681 aattcaagtg gtcattttga tattttacgc caaaattttt agcgttcctt tacttgataa
     4741 tttatccaaa aaccttcatt cgtcaaaaca attatgatat aattaattta tgatatgatt
     4801 aattaatatt tacggataac taatcatttc aatgcaatag atcaacatgc aaatcaatat
     4861 ccaccttact ccgaacattt tgcaccaacg aatgcccagc actatatccc ggcaagcaat
     4921 ataacgtcta taatttttta tgataacgat aaaattaaat tttatgaaag gggtaaaagc
     4981 atgagaaaaa tagctgtatt gttaattact ttgacgttct tagtcacgac tcgaatttag
     5041 gaggcttact tgtctgcttt cttcattaga atcaatcctt ttttaaaagt caatcccgtt
     5101 tgttgaacta ctctttaata aaataatttt tccgttccca attccacatt gcaataatag
     5161 aaaatccatc ttcatcggct ttttcgtcat catctgtatg aatcaaatcg ccttcttctg
     5221 tgtcatcaag gtttaatttt ttatgtattt cttttaacaa accaccatag gagattaacc
     5281 ttttacggtg taaaccttcc tccaaatcag acaaacgttt caaattcttt tcttcatcat
     5341 cggtcataaa atccgtatcc tttacaggat attttgcagt ttcgtcaatt gccgattgta
     5401 tatccgattt atatttattt ttcggtcgaa tcatttgaac ttttacattt ggatcatagt
     5461 ctaatttcat tgcctttttc caaaattgaa tccattgttt ttgattcacg tagttttctg
     5521 tattcttaaa ataagttggt tccacacata ccaatacatg catgtgctga ttataagaat
     5581 tatctttatt atttattgtc acttccgttg cacgcataaa accaacaaga tttttattaa
     5641 tttttttata ttgcatcatt cggcgaaatc cttgagccat atctgacaaa ctcttattta
     5701 attcttcgcc atcataaaca tttttaactg ttaatgtgag aaacaaccaa cgaactgttg
     5761 gcttttgttt aataacttca gcaacaacct tttgtgactg aatgccatgt ttcattgctc
     5821 tcctccagtt gcacattgga caaagcctgg atttacaaaa ccacactcga tacaactttc
     5881 tttcgcctgt ttcacgattt tgtttatact ctaatatttc agcacaatct tttactcttt
     5941 cagccttttt aaattcaaga atatgcagaa gttcaaagta atcaacatta gcgattttct
     6001 tttctctcca tggtctcact tttccacttt ttgtcttgtc cactaaaacc cttgattttt
     6061 catctgaata aatgctacta ttaggacaca taatattaaa agaaaccccc atctatttag
     6121 ttatttgttt ggtcacttat aactttaaca gatggggttt ttctgtgcaa ccaattttaa
     6181 gggttttcaa tactttaaaa cacatacata ccaacacttc aacgcacctt tcagcaacta
     6241 aaataaaaat gacgttattt ctatatgtat caagaataga aagaactcgt ttttcgctac
     6301 gctcaaaacg caaaaaaagc actcattcga gtgctttttc ttatcgctcc aaatcatgcg
     6361 attttttcct ctttgctttt ctttgctcac gaagttctcg atcacgctgc aaaacatctt
     6421 gaagcgaaaa agtattcttc ttttcttccg atcgctcatg ctgacgcacg aaaagccctc
     6481 taggcgcata ggaacaactc ctaaatgcat gtgaggggtt ttctcgtcca tgtgaacagt
     6541 cgcatacgca atattttgtt tcccatagcg gccgcgatta tcaaaaagga tcttcaccta
     6601 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6661 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6721 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agcgcttacc
     6781 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6841 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6901 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6961 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7021 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7081 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7141 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7201 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7261 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7321 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7381 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7441 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7501 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatggg taataactga
     7561 tataattaaa ttgaagctct aatttgtgag tttagtatac atgcatttac ttataataca
     7621 gttttttagt tttgctggcc gcatcttctc aaatatgctt cccagcctgc ttttctgtaa
     7681 cgttcaccct ctaccttagc atcccttccc tttgcaaata gtcctcttcc aacaataata
     7741 atgtcagatc ctgtagagac cacatcatcc acggttctat actgttgacc caatgcgtct
     7801 cccttgtcat ctaaacccac accgggtgtc ataatcaacc aatcgtaacc ttcatctctt
     7861 ccacccatgt ctctttgagc aataaagccg ataacaaaat ctttgtcgct cttcgcaatg
     7921 tcaacagtac ccttagtata ttctccagta gatagggagc ccttgcatga caattctgct
     7981 aacatcaaaa ggcctctagg ttcctttgtt acttcttctg ccgcctgctt caaaccgcta
     8041 acaatacctg ggcccaccac accgtgtgca ttcgtaatgt ctgcccattc tgctattctg
     8101 tatacacccg cagagtactg caatttgact gtattaccaa tgtcagcaaa ttttctgtct
     8161 tcgaagagta aaaaattgta cttggcggat aatgccttta gcggcttaac tgtgccctcc
     8221 atggaaaaat cagtcaagat atccacatgt gtttttagta aacaaatttt gggacctaat
     8281 gcttcaacta actccagtaa ttccttggtg gtacgaacat ccaatgaagc acacaagttt
     8341 gtttgctttt cgtgcatgat attaaatagc ttggcagcaa caggactagg atgagtagca
     8401 gcacgttcct tatatgtagc tttcgacatg atttatcttc gtttcctgca ggtttttgtt
     8461 ctgtgcagtt gggttaagaa tactgggcaa tttcatgttt cttcaacact acatatgcgt
     8521 atatatacca atctaagtct gtgctccttc cttcgttctt ccttctgttc ggagattacc
     8581 gaatcaaaaa aatttcaagg aaaccgaaat caaaaaaaag aataaaaaaa aaatgatgaa
     8641 ttgaaaagct agcttatcga tg
//
