LOCUS       V008004                 7023 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V008004
VERSION     V008004
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7023)
  AUTHORS   Olson DG.
  TITLE     Conversion of phosphoenolpyruvate to pyruvate in Clostridium
            thermocellum
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7023)
  AUTHORS   Olson DG.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-FEB-2016) Thayer School of Engineering, Dartmouth
            College, 14 Engineering Drive, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 7023)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7023)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (05-FEB-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..7023
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..455)
                     /direction=LEFT
                     /label="C. thermocellum replication origin"
                     /note="C. thermocellum replication origin"
     CDS             complement(561..1418)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(1419..1523)
                     /label="AmpR promoter"
     CDS             2191..2838
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     CDS             2859..3404
                     /codon_start=1
                     /gene="hpt"
                     /product="hypoxanthine phosphoribosyltransferase"
                     /label="hpt"
                     /note="Hpt; counterselection with 8AZH"
                     /protein_id="AOC59180.1"
                     /translation="MENLSKDIDEILITEEELKEKIKELGRQITKDYKGKNLMLVGVLK
                     GALMFMADLSRHIDLPLSLDFMAVSSYGSSTHSSGIVKIIKDLDISIEGKDVLIVEDII
                     DSGLTLSYLRETLLGRKPKSLKICTILDKPERREASVKVDYVGFKIPDKFVVGYGLDFD
                     EKYRNLPFIGVLKPEMYS"
     gene            2859..3404
                     /gene="hpt"
                     /label="hpt"
     terminator      3621..3809
                     /label="CYC1 terminator"
                     /note="transcription terminator for CYC1"
     rep_origin      complement(4069..4657)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(4744..5322)
                     /codon_start=1
                     /gene="tdk"
                     /product="thymidine kinase"
                     /label="tdk"
                     /note="Tdk; marker for 5FOA counter selection"
                     /protein_id="AOC59182.1"
                     /translation="MYGPKDHGYIEVVTGPMFSGKSEELIRRIKRAKIARQKVQVFKPA
                     IDDRYSIDKVVSHNGDNMHAIAIVKASDILAYAEEDTDVFAIDEVQFFDSEIVDIVKEI
                     ADSGKRVICAGLDMDFRGEPFGPTPELMAIAEFVDKLTAICMKCGNPATRTQRLINGKP
                     ANYDDPIIMVGAKESYEARCRKCHEVPRT"
     gene            complement(4744..5322)
                     /gene="tdk"
                     /label="tdk"
     CDS             complement(6022..7023)
                     /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 atatcgtggg
     1621 aataggcatg gaaaagattt ttttgcccga ggatgcattg ataaagtatt tcttttccga
     1681 aagagaggaa aagattctaa agagttttgg aaatactgat gaatattgtg tgcagagtac
     1741 aattctatgg acaagaaaag aggctttgtc aaaacttttt cgtctgggaa tgaggatgga
     1801 ttttaaaaag ctggatactt tggagggcga ggtggttttt caggaaacaa acagggcgcg
     1861 tctgttttct tttatatgca ataattactg tatctctctg gcattgccag gttttaataa
     1921 agattaaaat tattgactag aaataaaaaa attgtccata atattaatgg acaaaaaaac
     1981 aaagaattac atcaaaggaa gataaaaata ctttgttaaa aaattaatta ttttttatct
     2041 aaactattga aaatgaaaat aaaataatat aaaatgaatc atagtgcaag agatacttgc
     2101 cagaggatga atattttact gcattcatgc tttatggcag ctaatagagg cattaaatta
     2161 aattttaatt tacaatagga ggcgatatta atgaacttta ataaaattga tttagacaat
     2221 tggaagagaa aagagatatt taatcattat ttgaaccaac aaacgacttt tagtataacc
     2281 acagaaattg atattagtgt tttataccga aacataaaac aagaaggata taaattttac
     2341 cctgcattta ttttcttagt gacaagggtg ataaactcaa atacagcttt tagaactggt
     2401 tacaatagcg acggagagtt aggttattgg gataagttag agccacttta tacaattttt
     2461 gatggtgtat ctaaaacatt ctctggtatt tggactcctg taaagaatga cttcaaagag
     2521 ttttatgatt tatacctttc tgatgtagag aaatataatg gttcggggaa attgtttccc
     2581 aaaacaccta tacctgaaaa tgctttttct ctttctatta ttccatggac ttcatttact
     2641 gggtttaact taaatatcaa taataatagt aattaccttc tacccattat tacagcagga
     2701 aaattcatta ataaaggtaa ttcaatatat ttaccgctat ctttacaggt acatcattct
     2761 gtttgtgatg gttatcatgc aggattgttt atgaactcta ttcaggaatt gtcagatagg
     2821 cctaatgact ggcttttata ataaaggagg tcgacgtcat ggaaaattta tcaaaagaca
     2881 tcgatgaaat tttgatcaca gaagaagaac ttaaggaaaa gataaaagag cttgggaggc
     2941 aaatcacaaa agactacaaa gggaaaaatt tgatgttggt aggagtttta aaaggtgctt
     3001 taatgtttat ggctgatttg tcaagacaca tagatttgcc tttatcactt gattttatgg
     3061 ctgtttccag ctatggaagc tcaactcatt catcaggaat agtaaagata atcaaagatc
     3121 ttgatataag catagaaggc aaagatgttc tgattgtgga agacataatt gacagcggtt
     3181 tgactttgtc ttacttaagg gaaactttac ttggaaggaa gccaaaaagc ctgaaaatat
     3241 gcacaatatt agacaaaccg gagagaagag aagcatctgt aaaagtcgat tatgtaggat
     3301 ttaagatacc tgataagttt gtcgtgggtt atggattgga ctttgatgaa aagtacagga
     3361 accttccttt tataggcgtt ttgaaacctg aaatgtacag ctaagcccgg gcctcgagaa
     3421 aacaaaaggc tcagtcggaa gactgggcct tttgttttgg taccgaattc ggcgcgcctc
     3481 agcgtttaaa cccgctgatc ctagagggcc gcatcatgta attagttatg tcacgcttac
     3541 attcacgccc tccccatgct ttaatacata cctggcccag tagttcagct gctaatagta
     3601 gtgaaaaaat cagggataat gccgctctaa ccgaaaagga aggagttaga caacctgaag
     3661 tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta tttatatttc
     3721 aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata ctgaaaacct
     3781 tgcttgagaa ggttttggga cgctcgaagg ctttaatttg caagctgcgg ccctgcatta
     3841 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     3901 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     3961 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4021 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4081 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4141 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4201 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4261 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4321 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4381 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4441 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4501 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4561 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4621 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     4681 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgaaaaaatc
     4741 cgcttaagtc cgcgggactt catggcactt tctacacctt gcttcataag actcctttgc
     4801 tccaaccatt ataatgggat cgtcgtaatt ggcaggcttc ccatttataa gcctttgcgt
     4861 gcgagtagca ggattgccac acttcatgca tatagcagta agcttatcga caaattcagc
     4921 tatggccatc aattctggag tcggaccaaa tggttcacct ctaaagtcca tgtcaagccc
     4981 tgcacatatg actcttttcc cgctatcggc aatctctttt acgatgtcga ctatttcaga
     5041 atcaaaaaat tgaacttcat ctatagcaaa tacatccgta tcttcttcag cataagccaa
     5101 tatgtcagaa gcctttacta tggcaatggc gtgcatgttg tcgccgttat gagatacgac
     5161 tttgtctatg gaatacctat cgtctatagc cggtttgaaa acctgaactt tttgcctggc
     5221 aatcttagct ctctttatcc ttcttataag ttcctcactt tttccactga acatgggacc
     5281 agttacaact tctatgtacc cgtggtcttt aggcccatac atctttcgtc ctccttaaaa
     5341 ttttcgttta tttgtttttt atttagtgtg cctaaaagca ccatcttata cattcgtacc
     5401 ttatatctct tatcttgtat ctcttacctc atatctctta ttattaacta ccgaagtata
     5461 ttttcataac ggaatcttat gacttatttt tctgactgcc gttttaccat acggttaatt
     5521 tatacggaaa cagctaatct aaccttgctg aaggcatgtt taccaaaata ttatatctac
     5581 tttaccatat taattcaagt ggtcattttg atattttacg ccaaaatttt tagcgttcct
     5641 ttacttgata atttatccaa aaaccttcat tcgtcaaaac aattatgata taattaattt
     5701 atgatatgat taattaatat ttacggataa ctaatcattt caatgcaata gatcaacatg
     5761 caaatcaata tccaccttac tccgaacatt ttgcaccaac gaatgcccag cactatatcc
     5821 cggcaagcaa tataacgtct ataatttttt atgataacga taaaattaaa ttttatgaaa
     5881 ggggtaaaag catgagaaaa atagctgtat tgttaattac tttgacgttc ttagtcacga
     5941 ctcgaattta ggaggcttac ttgtctgctt tcttcattag aatcaatcct tttttaaaag
     6001 tcaatcccgt ttgttgaact actctttaat aaaataattt ttccgttccc aattccacat
     6061 tgcaataata gaaaatccat cttcatcggc tttttcgtca tcatctgtat gaatcaaatc
     6121 gccttcttct gtgtcatcaa ggtttaattt tttatgtatt tcttttaaca aaccaccata
     6181 ggagattaac cttttacggt gtaaaccttc ctccaaatca gacaaacgtt tcaaattctt
     6241 ttcttcatca tcggtcataa aatccgtatc ctttacagga tattttgcag tttcgtcaat
     6301 tgccgattgt atatccgatt tatatttatt tttcggtcga atcatttgaa cttttacatt
     6361 tggatcatag tctaatttca ttgccttttt ccaaaattga atccattgtt tttgattcac
     6421 gtagttttct gtattcttaa aataagttgg ttccacacat accaatacat gcatgtgctg
     6481 attataagaa ttatctttat tatttattgt cacttccgtt gcacgcataa aaccaacaag
     6541 atttttatta atttttttat attgcatcat tcggcgaaat ccttgagcca tatctgacaa
     6601 actcttattt aattcttcgc catcataaac atttttaact gttaatgtga gaaacaacca
     6661 acgaactgtt ggcttttgtt taataacttc agcaacaacc ttttgtgact gaatgccatg
     6721 tttcattgct ctcctccagt tgcacattgg acaaagcctg gatttacaaa accacactcg
     6781 atacaacttt ctttcgcctg tttcacgatt ttgtttatac tctaatattt cagcacaatc
     6841 ttttactctt tcagcctttt taaattcaag aatatgcaga agttcaaagt aatcaacatt
     6901 agcgattttc ttttctctcc atggtctcac ttttccactt tttgtcttgt ccactaaaac
     6961 ccttgatttt tcatctgaat aaatgctact attaggacac ataatattaa aagaaacccc
     7021 cat
//
