LOCUS       V001906                 8248 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V001906
VERSION     V001906
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8248)
  AUTHORS   Deng Y, Olson DG, Zhou J, Herring CD, Joe Shaw A, Lynd LR.
  TITLE     Redirecting carbon flux through exogenous pyruvate kinase to achieve
            high ethanol yields in Clostridium thermocellum
  JOURNAL   Metab. Eng. 15, 151-158 (2013)
   PUBMED   23202749
REFERENCE   2  (bases 1 to 8248)
  AUTHORS   Zhou J, Olson DG, Lynd LR.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-NOV-2013) Thayer School of Engineering, Dartmouth
            College, 14 Engineering Drive, Hanover, NH 03755, U.S
REFERENCE   3  (bases 1 to 8248)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8248)
  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.
            15, 151-158 (2013)"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (05-NOV-2013) Thayer School of Engineering, Dartmouth College, 14
            Engineering Drive, Hanover, NH 03755, U.S"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8248
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..455)
                     /direction=LEFT
                     /label="Clostridium thermocellum"
                     /note="Clostridium thermocellum"
     CDS             complement(561..1418)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(1419..1523)
                     /label="AmpR promoter"
     misc_feature    1613..2012
                     /label="similar to Clo1313_1879"
                     /note="similar to Clo1313_1879"
     regulatory      2016..2590
                     /gene="GAPDH"
                     /regulatory_class="promoter"
     gene            2016..2590
                     /gene="GAPDH"
                     /label="GAPDH"
     CDS             2591..3238
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     CDS             3259..3804
                     /codon_start=1
                     /locus_tag="or0940"
                     /product="hypoxanthine phosphoribosyl transferase"
                     /EC_number="2.4.2.8"
                     /label="or0940"
                     /protein_id="AHH35087.1"
                     /translation="MENLSKDIDEILITEEELKEKIKELGRQITKDYKGKNLMLVGVLK
                     GALMFMADLSRHIDLPLSLDFMAVSSYGSSTHSSGIVKIIKDLDISIEGKDVLIVEDII
                     DSGLTLSYLRETLLGRKPKSLKICTILDKPERREASVKVDYVGFKIPDKFVVGYGLDFD
                     EKYRNLPFIGVLKPEMYS"
     gene            3259..3804
                     /locus_tag="or0940"
                     /label="or0940"
     regulatory      3819..3858
                     /label="hairpin"
                     /note="hairpin"
                     /regulatory_class="terminator"
     misc_feature    4391..4813
                     /label="similar to Clo1313_1879"
                     /note="similar to Clo1313_1879"
     terminator      4846..5034
                     /label="CYC1 terminator"
                     /note="transcription terminator for CYC1"
     rep_origin      complement(5294..5882)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5969..6547)
                     /codon_start=1
                     /gene="tdk"
                     /locus_tag="or0326"
                     /product="thymidine kinase"
                     /EC_number="2.7.1.21"
                     /label="tdk"
                     /protein_id="AHH35090.1"
                     /translation="MYGPKDHGYIEVVTGPMFSGKSEELIRRIKRAKIARQKVQVFKPA
                     IDDRYSIDKVVSHNGDNMHAIAIVKASDILAYAEEDTDVFAIDEVQFFDSEIVDIVKEI
                     ADSGKRVICAGLDMDFRGEPFGPTPELMAIAEFVDKLTAICMKCGNPATRTQRLINGKP
                     ANYDDPIIMVGAKESYEARCRKCHEVPRT"
     gene            complement(5969..6547)
                     /gene="tdk"
                     /locus_tag="or0326"
                     /label="tdk"
     regulatory      complement(6548..7168)
                     /gene="cbp"
                     /regulatory_class="promoter"
     gene            complement(6548..7168)
                     /gene="cbp"
                     /label="cbp"
     CDS             complement(7247..8248)
                     /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 atgaagagtt
     1621 gtcgcttgct tatacaccgg gtgttgcaga accatgtctt gcaattcaga aagatgttaa
     1681 tctttcttat gaatatacaa gacgttggaa cctggtagcg gttattaccg acggtacggc
     1741 ggttttaggg ctcggagaca taggacctga agccggaatg cctgttatgg aaggtaaatg
     1801 cgtactcttc aagaggtttg gtgatgtgga cgcatttccg ctctgtatca aatcaaaaga
     1861 cgtagatgaa attgtaaaga caatcaagct catctccgga agctttggcg gtataaacct
     1921 cgaagatata tccgctccga gatgctttga aatagaaaga agactcaaag aggaatgtga
     1981 cattccaata ttccatgatg accagcacgg taatcgtggg aataggcatg gaaaagattt
     2041 ttttgcccga ggatgcattg ataaagtatt tcttttccga aagagaggaa aagattctaa
     2101 agagttttgg aaatactgat gaatattgtg tgcagagtac aattctatgg acaagaaaag
     2161 aggctttgtc aaaacttttt cgtctgggaa tgaggatgga ttttaaaaag ctggatactt
     2221 tggagggcga ggtggttttt caggaaacaa acagggcgcg tctgttttct tttatatgca
     2281 ataattactg tatctctctg gcattgccag gttttaataa agattaaaat tattgactag
     2341 aaataaaaaa attgtccata atattaatgg acaaaaaaac aaagaattac atcaaaggaa
     2401 gataaaaata ctttgttaaa aaattaatta ttttttatct aaactattga aaatgaaaat
     2461 aaaataatat aaaatgaatc atagtgcaag agatacttgc cagaggatga atattttact
     2521 gcattcatgc tttatggcag ctaatagagg cattaaatta aattttaatt tacaatagga
     2581 ggcgatatta atgaacttta ataaaattga tttagacaat tggaagagaa aagagatatt
     2641 taatcattat ttgaaccaac aaacgacttt tagtataacc acagaaattg atattagtgt
     2701 tttataccga aacataaaac aagaaggata taaattttac cctgcattta ttttcttagt
     2761 gacaagggtg ataaactcaa atacagcttt tagaactggt tacaatagcg acggagagtt
     2821 aggttattgg gataagttag agccacttta tacaattttt gatggtgtat ctaaaacatt
     2881 ctctggtatt tggactcctg taaagaatga cttcaaagag ttttatgatt tatacctttc
     2941 tgatgtagag aaatataatg gttcggggaa attgtttccc aaaacaccta tacctgaaaa
     3001 tgctttttct ctttctatta ttccatggac ttcatttact gggtttaact taaatatcaa
     3061 taataatagt aattaccttc tacccattat tacagcagga aaattcatta ataaaggtaa
     3121 ttcaatatat ttaccgctat ctttacaggt acatcattct gtttgtgatg gttatcatgc
     3181 aggattgttt atgaactcta ttcaggaatt gtcagatagg cctaatgact ggcttttata
     3241 ataaaggagg tcgacgtcat ggaaaattta tcaaaagaca tcgatgaaat tttgatcaca
     3301 gaagaagaac ttaaggaaaa gataaaagag cttgggaggc aaatcacaaa agactacaaa
     3361 gggaaaaatt tgatgttggt aggagtttta aaaggtgctt taatgtttat ggctgatttg
     3421 tcaagacaca tagatttgcc tttatcactt gattttatgg ctgtttccag ctatggaagc
     3481 tcaactcatt catcaggaat agtaaagata atcaaagatc ttgatataag catagaaggc
     3541 aaagatgttc tgattgtgga agacataatt gacagcggtt tgactttgtc ttacttaagg
     3601 gaaactttac ttggaaggaa gccaaaaagc ctgaaaatat gcacaatatt agacaaaccg
     3661 gagagaagag aagcatctgt aaaagtcgat tatgtaggat ttaagatacc tgataagttt
     3721 gtcgtgggtt atggattgga ctttgatgaa aagtacagga accttccttt tataggcgtt
     3781 ttgaaacctg aaatgtacag ctaagcccgg gcctcgagaa aacaaaaggc tcagtcggaa
     3841 gactgggcct tttgttttgg taccgaattc ggcgcgcctc agcgtttaaa cccgctgatc
     3901 ctagagggcc gcatcatgta attagttatg tcacgcttac attcacgccc tccccatgct
     3961 ttaatacata cctggcccag tagttcaggc tacataatag gcgaacacgg tgattcacag
     4021 cttccgttgt ggagctgcac acatatcgcc ggtaaaaata tcaacgaata tatcgatgat
     4081 ccgaaatgca atttcacaga agaagacaag aaaaaaatcg ctgaagatgt taaaactgcg
     4141 ggtgcaacca ttatcaagaa caaaggtgca acatactatg gtattgcagt ttcaatcaac
     4201 acaatagttg aaacactcct taagaatcag aatacaataa gaaccgtagg aaccgttata
     4261 aacggcatgt atggaataga agatgttgca ataagccttc catccatcgt aaattccgaa
     4321 ggtgttcagg aagttctcca atttaatctg actcctgaag aagaagaagc tttaagattc
     4381 tcagcggagc ggaagagaca acttaaacag tgaaaaagcc ctgattgcta aaatctcgaa
     4441 cctcgagaaa aagaaaggta ctcttgaaga tgtaatcaag ggagctgacg tattcatcgg
     4501 tctttccgtt ccaggaacag ttacaaagga tatggtaaaa tccatggcaa aggatccgat
     4561 tatctttgct atggcaaatc ctactcctga aataatgcct gatgaagcaa aagaagcagg
     4621 agcaaaggta gtgggtaccg gaagatccga cttcccgaac cagataaaca acgttcttgc
     4681 gttccccgga atattcagag gtgcgcttga tgtaagagca agagatatca atgatgaaat
     4741 gaagatagcc gctgcaaaag caatagcttc tctggtaagc gatgaagagc tcaatcctga
     4801 cttcattctt ccgctgctaa tagtagtgaa aaaatcaggg ataatgccgc tctaaccgaa
     4861 aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttat agttatgtta
     4921 gtattaagaa cgttatttat atttcaaatt tttctttttt ttctgtacag acgcgtgtac
     4981 gcatgtaaca ttatactgaa aaccttgctt gagaaggttt tgggacgctc gaaggcttta
     5041 atttgcaagc tgcggccctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     5101 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5161 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5221 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5281 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5341 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5401 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5461 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5521 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5581 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5641 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5701 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     5761 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5821 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5881 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5941 aagggatttt ggtcatgaaa aaatccgctt aagtccgcgg gacttcatgg cactttctac
     6001 accttgcttc ataagactcc tttgctccaa ccattataat gggatcgtcg taattggcag
     6061 gcttcccatt tataagcctt tgcgtgcgag tagcaggatt gccacacttc atgcatatag
     6121 cagtaagctt atcgacaaat tcagctatgg ccatcaattc tggagtcgga ccaaatggtt
     6181 cacctctaaa gtccatgtca agccctgcac atatgactct tttcccgcta tcggcaatct
     6241 cttttacgat gtcgactatt tcagaatcaa aaaattgaac ttcatctata gcaaatacat
     6301 ccgtatcttc ttcagcataa gccaatatgt cagaagcctt tactatggca atggcgtgca
     6361 tgttgtcgcc gttatgagat acgactttgt ctatggaata cctatcgtct atagccggtt
     6421 tgaaaacctg aactttttgc ctggcaatct tagctctctt tatccttctt ataagttcct
     6481 cactttttcc actgaacatg ggaccagtta caacttctat gtacccgtgg tctttaggcc
     6541 catacatctt tcgtcctcct taaaattttc gtttatttgt tttttattta gtgtgcctaa
     6601 aagcaccatc ttatacattc gtaccttata tctcttatct tgtatctctt acctcatatc
     6661 tcttattatt aactaccgaa gtatattttc ataacggaat cttatgactt atttttctga
     6721 ctgccgtttt accatacggt taatttatac ggaaacagct aatctaacct tgctgaaggc
     6781 atgtttacca aaatattata tctactttac catattaatt caagtggtca ttttgatatt
     6841 ttacgccaaa atttttagcg ttcctttact tgataattta tccaaaaacc ttcattcgtc
     6901 aaaacaatta tgatataatt aatttatgat atgattaatt aatatttacg gataactaat
     6961 catttcaatg caatagatca acatgcaaat caatatccac cttactccga acattttgca
     7021 ccaacgaatg cccagcacta tatcccggca agcaatataa cgtctataat tttttatgat
     7081 aacgataaaa ttaaatttta tgaaaggggt aaaagcatga gaaaaatagc tgtattgtta
     7141 attactttga cgttcttagt cacgactcga atttaggagg cttacttgtc tgctttcttc
     7201 attagaatca atcctttttt aaaagtcaat cccgtttgtt gaactactct ttaataaaat
     7261 aatttttccg ttcccaattc cacattgcaa taatagaaaa tccatcttca tcggcttttt
     7321 cgtcatcatc tgtatgaatc aaatcgcctt cttctgtgtc atcaaggttt aattttttat
     7381 gtatttcttt taacaaacca ccataggaga ttaacctttt acggtgtaaa ccttcctcca
     7441 aatcagacaa acgtttcaaa ttcttttctt catcatcggt cataaaatcc gtatccttta
     7501 caggatattt tgcagtttcg tcaattgccg attgtatatc cgatttatat ttatttttcg
     7561 gtcgaatcat ttgaactttt acatttggat catagtctaa tttcattgcc tttttccaaa
     7621 attgaatcca ttgtttttga ttcacgtagt tttctgtatt cttaaaataa gttggttcca
     7681 cacataccaa tacatgcatg tgctgattat aagaattatc tttattattt attgtcactt
     7741 ccgttgcacg cataaaacca acaagatttt tattaatttt tttatattgc atcattcggc
     7801 gaaatccttg agccatatct gacaaactct tatttaattc ttcgccatca taaacatttt
     7861 taactgttaa tgtgagaaac aaccaacgaa ctgttggctt ttgtttaata acttcagcaa
     7921 caaccttttg tgactgaatg ccatgtttca ttgctctcct ccagttgcac attggacaaa
     7981 gcctggattt acaaaaccac actcgataca actttctttc gcctgtttca cgattttgtt
     8041 tatactctaa tatttcagca caatctttta ctctttcagc ctttttaaat tcaagaatat
     8101 gcagaagttc aaagtaatca acattagcga ttttcttttc tctccatggt ctcacttttc
     8161 cactttttgt cttgtccact aaaacccttg atttttcatc tgaataaatg ctactattag
     8221 gacacataat attaaaagaa acccccat
//
