LOCUS       V008002                 9473 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V008002
VERSION     V008002
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9473)
  AUTHORS   Olson DG.
  TITLE     Conversion of phosphoenolpyruvate to pyruvate in Clostridium
            thermocellum
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9473)
  AUTHORS   Olson DG.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-FEB-2016) Thayer School of Engineering, Dartmouth
            College, 14 Engineering Drive, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 9473)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9473)
  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            "Unpublished"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
            (06-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..9473
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(4..1005)
                     /label="repB"
                     /note="RepB replication protein"
     rep_origin      complement(1006..1460)
                     /direction=LEFT
                     /label="C. thermocellum replication origin"
                     /note="C. thermocellum replication origin"
     CDS             complement(1566..2423)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(2424..2528)
                     /label="AmpR promoter"
     CDS             4857..5504
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     CDS             5525..6070
                     /codon_start=1
                     /gene="hpt"
                     /product="hypoxanthine phosphoribosyltransferase"
                     /label="hpt"
                     /note="Hpt; counterselection with 8AZH"
                     /protein_id="AOC59192.1"
                     /translation="MENLSKDIDEILITEEELKEKIKELGRQITKDYKGKNLMLVGVLK
                     GALMFMADLSRHIDLPLSLDFMAVSSYGSSTHSSGIVKIIKDLDISIEGKDVLIVEDII
                     DSGLTLSYLRETLLGRKPKSLKICTILDKPERREASVKVDYVGFKIPDKFVVGYGLDFD
                     EKYRNLPFIGVLKPEMYS"
     gene            5525..6070
                     /gene="hpt"
                     /label="hpt"
     misc_feature    6255..7043
                     /note="partial pyruvate kinase gene from T.
                     saccharolyticum"
     terminator      7076..7264
                     /label="CYC1 terminator"
                     /note="transcription terminator for CYC1"
     rep_origin      complement(7524..8112)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(8199..8777)
                     /codon_start=1
                     /gene="tdk"
                     /product="thymidine kinase"
                     /label="tdk"
                     /note="Tdk; counterselection with FUDR"
                     /protein_id="AOC59190.1"
                     /translation="MYGPKDHGYIEVVTGPMFSGKSEELIRRIKRAKIARQKVQVFKPA
                     IDDRYSIDKVVSHNGDNMHAIAIVKASDILAYAEEDTDVFAIDEVQFFDSEIVDIVKEI
                     ADSGKRVICAGLDMDFRGEPFGPTPELMAIAEFVDKLTAICMKCGNPATRTQRLINGKP
                     ANYDDPIIMVGAKESYEARCRKCHEVPRT"
     gene            complement(8199..8777)
                     /gene="tdk"
                     /label="tdk"
ORIGIN
        1 ctactcttta ataaaataat ttttccgttc ccaattccac attgcaataa tagaaaatcc
       61 atcttcatcg gctttttcgt catcatctgt atgaatcaaa tcgccttctt ctgtgtcatc
      121 aaggtttaat tttttatgta tttcttttaa caaaccacca taggagatta accttttacg
      181 gtgtaaacct tcctccaaat cagacaaacg tttcaaattc ttttcttcat catcggtcat
      241 aaaatccgta tcctttacag gatattttgc agtttcgtca attgccgatt gtatatccga
      301 tttatattta tttttcggtc gaatcatttg aacttttaca tttggatcat agtctaattt
      361 cattgccttt ttccaaaatt gaatccattg tttttgattc acgtagtttt ctgtattctt
      421 aaaataagtt ggttccacac ataccaatac atgcatgtgc tgattataag aattatcttt
      481 attatttatt gtcacttccg ttgcacgcat aaaaccaaca agatttttat taattttttt
      541 atattgcatc attcggcgaa atccttgagc catatctgac aaactcttat ttaattcttc
      601 gccatcataa acatttttaa ctgttaatgt gagaaacaac caacgaactg ttggcttttg
      661 tttaataact tcagcaacaa ccttttgtga ctgaatgcca tgtttcattg ctctcctcca
      721 gttgcacatt ggacaaagcc tggatttaca aaaccacact cgatacaact ttctttcgcc
      781 tgtttcacga ttttgtttat actctaatat ttcagcacaa tcttttactc tttcagcctt
      841 tttaaattca agaatatgca gaagttcaaa gtaatcaaca ttagcgattt tcttttctct
      901 ccatggtctc acttttccac tttttgtctt gtccactaaa acccttgatt tttcatctga
      961 ataaatgcta ctattaggac acataatatt aaaagaaacc cccatctatt tagttatttg
     1021 tttggtcact tataacttta acagatgggg tttttctgtg caaccaattt taagggtttt
     1081 caatacttta aaacacatac ataccaacac ttcaacgcac ctttcagcaa ctaaaataaa
     1141 aatgacgtta tttctatatg tatcaagaat agaaagaact cgtttttcgc tacgctcaaa
     1201 acgcaaaaaa agcactcatt cgagtgcttt ttcttatcgc tccaaatcat gcgatttttt
     1261 cctctttgct tttctttgct cacgaagttc tcgatcacgc tgcaaaacat cttgaagcga
     1321 aaaagtattc ttcttttctt ccgatcgctc atgctgacgc acgaaaagcc ctctaggcgc
     1381 ataggaacaa ctcctaaatg catgtgaggg gttttctcgt ccatgtgaac agtcgcatac
     1441 gcaatatttt gtttcccata gcggccgcga ttatcaaaaa ggatcttcac ctagatcctt
     1501 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     1561 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     1621 atagttgcct gactccccgt cgtgtagata actacgatac gggagcgctt accatctggc
     1681 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     1741 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     1801 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     1861 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     1921 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     1981 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     2041 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     2101 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     2161 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     2221 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     2281 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     2341 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     2401 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     2461 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     2521 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     2581 acattaacct ataaaaatag gcgtatcacg aggcgatgac actatcctgt atcctgattt
     2641 ttccgcaagg taaggaataa cggtatcatc aagcatcggc atcatctccg gcggaggacc
     2701 gggaagcatt accacaatct ttcccttatc ttcaataatg cagcccgggg ctgtgccgct
     2761 gttattctca accactgtgc acccttccgg aagatatgcc tgcttgacat tattgtccgt
     2821 catttttcgg tttatcctgg taaaaaaagt tttaatcctc tcaaggcttt cttcgtgtaa
     2881 aacaagcttt ttccccaaaa cttccgcaac agtctccttt gtaaggtcat cctgcgtggg
     2941 gccgagtcct ccggtcataa taacaaggtc gcacctttcc aaagctgcaa gaagacattt
     3001 tttcagccga acggaattgt cccccaccac actgtgataa tacacattca caccaatgtc
     3061 attgagcctt ttggatatat actgggcatt ggtatttgct atctgcccca ttaaaagctc
     3121 ggttccaacc gctaatatct ccgcattcat attgaaagac cccttaaatt taaacttttt
     3181 gtaacttatt atatcaatta gtgttataaa ataaaaggga aaaagaatta aaatcaaagg
     3241 tttcaagagc agccgtatca cccgtaaaag tttcagccga ttcaaccttt ttacacataa
     3301 aactttcaaa aattgatgac ttacaattat caagtaggat ataatattac taatgctaaa
     3361 cagttattga taaaggagga aggaataagt tctccttttc ttttatgaaa aggagaactt
     3421 tcattattga taaatatata aactagtata taattttaat ataaaaccta ttttacataa
     3481 tggaaattat ctatcggggg aggaaatatg aacaattcag tggaaatttt aaataaaatc
     3541 gtgtcaaata ttgaaaaagt cattgttgga aaaaagaaag ctatcgagtt gatattaata
     3601 tcacttattt gcgatggaca tgttttgatt gaagatgtcc ccggtgtcgg aaaaaccagt
     3661 attgtatcat ctcttgcaaa gtcggtaaat gcttctttta aaagaataca gtttacaccg
     3721 gatattcttc cttcagacat tacaggattt acgatgtaca atcagaaaga aggtaaattc
     3781 gaatatcatc ccggaagcat tatgagccag ataatacttg cggatgaaat aaaccgaaca
     3841 tctccaaaaa cccaggcaag ccttcttgaa gcaatggaag aaaaacaggt gaccgttgac
     3901 ggtgtgacat acaatcttcc aaggcctttt atggtactgg caactcaaaa tcccgtggag
     3961 tatctcggca cctatcctct tcccgaagct cagctggaca ggttttttat gaaagtttcc
     4021 ataggttatc ctgaaaagtt tgaagaatcc gaaatactgt caagatttca cgacgaaaac
     4081 cccttggaaa ccttaaaacc tgtggcggac agcagcgata tactgaatat tcaaagcgag
     4141 gttaaaaaag tttatgttga caggtccatc aacaactaca tagtggatat agtaagccgg
     4201 acaagattcc attcggaaat aagtctgggt tcaagtccaa gaggttctct ttctctatac
     4261 agagcctccc aggcatggat cgtgggaata ggcatggaaa agattttttt gcccgaggat
     4321 gcattgataa agtatttctt ttccgaaaga gaggaaaaga ttctaaagag ttttggaaat
     4381 actgatgaat attgtgtgca gagtacaatt ctatggacaa gaaaagaggc tttgtcaaaa
     4441 ctttttcgtc tgggaatgag gatggatttt aaaaagctgg atactttgga gggcgaggtg
     4501 gtttttcagg aaacaaacag ggcgcgtctg ttttctttta tatgcaataa ttactgtatc
     4561 tctctggcat tgccaggttt taataaagat taaaattatt gactagaaat aaaaaaattg
     4621 tccataatat taatggacaa aaaaacaaag aattacatca aaggaagata aaaatacttt
     4681 gttaaaaaat taattatttt ttatctaaac tattgaaaat gaaaataaaa taatataaaa
     4741 tgaatcatag tgcaagagat acttgccaga ggatgaatat tttactgcat tcatgcttta
     4801 tggcagctaa tagaggcatt aaattaaatt ttaatttaca ataggaggcg atattaatga
     4861 actttaataa aattgattta gacaattgga agagaaaaga gatatttaat cattatttga
     4921 accaacaaac gacttttagt ataaccacag aaattgatat tagtgtttta taccgaaaca
     4981 taaaacaaga aggatataaa ttttaccctg catttatttt cttagtgaca agggtgataa
     5041 actcaaatac agcttttaga actggttaca atagcgacgg agagttaggt tattgggata
     5101 agttagagcc actttataca atttttgatg gtgtatctaa aacattctct ggtatttgga
     5161 ctcctgtaaa gaatgacttc aaagagtttt atgatttata cctttctgat gtagagaaat
     5221 ataatggttc ggggaaattg tttcccaaaa cacctatacc tgaaaatgct ttttctcttt
     5281 ctattattcc atggacttca tttactgggt ttaacttaaa tatcaataat aatagtaatt
     5341 accttctacc cattattaca gcaggaaaat tcattaataa aggtaattca atatatttac
     5401 cgctatcttt acaggtacat cattctgttt gtgatggtta tcatgcagga ttgtttatga
     5461 actctattca ggaattgtca gataggccta atgactggct tttataataa aggaggtcga
     5521 cgtcatggaa aatttatcaa aagacatcga tgaaattttg atcacagaag aagaacttaa
     5581 ggaaaagata aaagagcttg ggaggcaaat cacaaaagac tacaaaggga aaaatttgat
     5641 gttggtagga gttttaaaag gtgctttaat gtttatggct gatttgtcaa gacacataga
     5701 tttgccttta tcacttgatt ttatggctgt ttccagctat ggaagctcaa ctcattcatc
     5761 aggaatagta aagataatca aagatcttga tataagcata gaaggcaaag atgttctgat
     5821 tgtggaagac ataattgaca gcggtttgac tttgtcttac ttaagggaaa ctttacttgg
     5881 aaggaagcca aaaagcctga aaatatgcac aatattagac aaaccggaga gaagagaagc
     5941 atctgtaaaa gtcgattatg taggatttaa gatacctgat aagtttgtcg tgggttatgg
     6001 attggacttt gatgaaaagt acaggaacct tccttttata ggcgttttga aacctgaaat
     6061 gtacagctaa gcccgggcct cgagaaaaca aaaggctcag tcggaagact gggccttttg
     6121 ttttggtacc gaattcggcg cgcctcagcg tttaaacccg ctgatcctag agggccgcat
     6181 catgtaatta gttatgtcac gcttacattc acgccctccc catgctttaa tacatacctg
     6241 gcccagtagt tcagcatgga agcagaatag acaatattat aaagattaga gaagaactta
     6301 agctgcctat tgcaattatg cttgatacaa aagggcctga aataaggact ggcagattta
     6361 aaggcggtgt tgcagagctt aaagaaggcc agacatttac gataacatca agggaaattg
     6421 aaggagataa cactatttgt tctgtttcat acaaggggct tcctcaagat gtggagagag
     6481 gttctcgcat attgattgat gacggattag tatcattgaa agtcaatgac gtaaaaggtg
     6541 aagatatagt atgcactgtg gagaattctg gtacaatagg tgatcacaaa ggtgtaaatg
     6601 tacctggtac aaagcttaat ttgcctgcca taacgcaaaa agacgtggat gatatagagt
     6661 ttggaataaa aaaaggaatc gacatgattg cagcgtcttt tgtcagaaaa gcagcagatg
     6721 taattgccat aaggagattg ttagaagaca atgacgctgg ccatatactt atcatatcaa
     6781 aaattgaaaa tcgcgaaggc gtagaaaata ttgacgaaat aatcaaagtc tctgatggca
     6841 taatggtagc ccgcggcgat ttgggtgtcg aaattcctat agaggaaata cctatcgttc
     6901 agaaaaggat aattgaaaaa tgcaacaaag caggtaaacc agtagttact gctacacaga
     6961 tgcttgactc tatgataaga aatccaaggc caacaagggc agaagtaaca gatgtagcca
     7021 atgctatatt ggatggcact gatctgctaa tagtagtgaa aaaatcaggg ataatgccgc
     7081 tctaaccgaa aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttat
     7141 agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt ttctgtacag
     7201 acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt gagaaggttt tgggacgctc
     7261 gaaggcttta atttgcaagc tgcggccctg cattaatgaa tcggccaacg cgcggggaga
     7321 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     7381 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     7441 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     7501 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     7561 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     7621 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     7681 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     7741 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     7801 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     7861 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     7921 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     7981 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     8041 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     8101 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     8161 aactcacgtt aagggatttt ggtcatgaaa aaatccgctt aagtccgcgg gacttcatgg
     8221 cactttctac accttgcttc ataagactcc tttgctccaa ccattataat gggatcgtcg
     8281 taattggcag gcttcccatt tataagcctt tgcgtgcgag tagcaggatt gccacacttc
     8341 atgcatatag cagtaagctt atcgacaaat tcagctatgg ccatcaattc tggagtcgga
     8401 ccaaatggtt cacctctaaa gtccatgtca agccctgcac atatgactct tttcccgcta
     8461 tcggcaatct cttttacgat gtcgactatt tcagaatcaa aaaattgaac ttcatctata
     8521 gcaaatacat ccgtatcttc ttcagcataa gccaatatgt cagaagcctt tactatggca
     8581 atggcgtgca tgttgtcgcc gttatgagat acgactttgt ctatggaata cctatcgtct
     8641 atagccggtt tgaaaacctg aactttttgc ctggcaatct tagctctctt tatccttctt
     8701 ataagttcct cactttttcc actgaacatg ggaccagtta caacttctat gtacccgtgg
     8761 tctttaggcc catacatctt tcgtcctcct taaaattttc gtttatttgt tttttattta
     8821 gtgtgcctaa aagcaccatc ttatacattc gtaccttata tctcttatct tgtatctctt
     8881 acctcatatc tcttattatt aactaccgaa gtatattttc ataacggaat cttatgactt
     8941 atttttctga ctgccgtttt accatacggt taatttatac ggaaacagct aatctaacct
     9001 tgctgaaggc atgtttacca aaatattata tctactttac catattaatt caagtggtca
     9061 ttttgatatt ttacgccaaa atttttagcg ttcctttact tgataattta tccaaaaacc
     9121 ttcattcgtc aaaacaatta tgatataatt aatttatgat atgattaatt aatatttacg
     9181 gataactaat catttcaatg caatagatca acatgcaaat caatatccac cttactccga
     9241 acattttgca ccaacgaatg cccagcacta tatcccggca agcaatataa cgtctataat
     9301 tttttatgat aacgataaaa ttaaatttta tgaaaggggt aaaagcatga gaaaaatagc
     9361 tgtattgtta attactttga cgttcttagt cacgactcga atttaggagg cttacttgtc
     9421 tgctttcttc attagaatca atcctttttt aaaagtcaat cccgtttgtt gaa
//
