LOCUS       V001845                 9005 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V001845
VERSION     V001845
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9005)
  AUTHORS   Zhou J, Olson DG.
  TITLE     Conversion of phosphoenolpyruvate to pyruvate in Clostridium
            thermocellum
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9005)
  AUTHORS   Zhou J, 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 9005)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9005)
  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            "Unpublished"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..9005
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(4..861)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(862..966)
                     /label="AmpR promoter"
     CDS             2986..3633
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     CDS             3654..4199
                     /codon_start=1
                     /gene="hpt"
                     /product="hypoxanthine phosphoribosyltransferase"
                     /label="hpt"
                     /note="Hpt; counterselection with 8AZH"
                     /protein_id="AOC59185.1"
                     /translation="MENLSKDIDEILITEEELKEKIKELGRQITKDYKGKNLMLVGVLK
                     GALMFMADLSRHIDLPLSLDFMAVSSYGSSTHSSGIVKIIKDLDISIEGKDVLIVEDII
                     DSGLTLSYLRETLLGRKPKSLKICTILDKPERREASVKVDYVGFKIPDKFVVGYGLDFD
                     EKYRNLPFIGVLKPEMYS"
     gene            3654..4199
                     /gene="hpt"
                     /label="hpt"
     terminator      5046..5234
                     /label="CYC1 terminator"
                     /note="transcription terminator for CYC1"
     rep_origin      complement(5494..6082)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6169..6747)
                     /codon_start=1
                     /gene="tdk"
                     /product="thymidine kinase"
                     /label="tdk"
                     /note="Tdk; counterselection with FUDR"
                     /protein_id="AOC59186.1"
                     /translation="MYGPKDHGYIEVVTGPMFSGKSEELIRRIKRAKIARQKVQVFKPA
                     IDDRYSIDKVVSHNGDNMHAIAIVKASDILAYAEEDTDVFAIDEVQFFDSEIVDIVKEI
                     ADSGKRVICAGLDMDFRGEPFGPTPELMAIAEFVDKLTAICMKCGNPATRTQRLINGKP
                     ANYDDPIIMVGAKESYEARCRKCHEVPRT"
     gene            complement(6169..6747)
                     /gene="tdk"
                     /label="tdk"
     CDS             complement(7447..8448)
                     /label="repB"
                     /note="RepB replication protein"
     rep_origin      complement(8449..8903)
                     /direction=LEFT
                     /label="C. thermocellum replication origin"
                     /note="C. thermocellum replication origin"
ORIGIN
        1 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
       61 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagcgcttac catctggccc
      121 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
      181 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
      241 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
      301 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
      361 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
      421 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
      481 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
      541 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
      601 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
      661 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
      721 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
      781 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
      841 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
      901 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
      961 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     1021 attaacctat aaaaataggc gtatcacgag gcgatccatc ttttagtctc tcttcgaaca
     1081 tataattatg aatgcctcaa aaataatcga gttttaagcc gtcttattga ttattataaa
     1141 cgcttgtata cccatataaa tagtgaatat gaccatcctc aaaagaggtt gcagtgtagt
     1201 aatagtgtat atgtccaccg ctaacgggta tactgggacc tgtttgtagg ctataacgat
     1261 gtatatgacc gtcttcgaag gtggtttcgc cagccatgta gtgtatgtga ccgggactgt
     1321 tgaggtttac actggttata cctgaatatt catgaatatg gccgtcatta aaagtagttc
     1381 ttcccataaa tctatgactg tgcaaactgt tttgcataat aatcacctca atacatatta
     1441 tgttaactat gtagagggtg ttacttctat aataacgtat tttatggtgc attaagcagt
     1501 aaataccctt aagttaacca ttgtagcgag atgctcaatg gaatggggaa aattcaattt
     1561 tttatgagat ttcttggtat tttttctgag cgttatatta taataaattt ataaaaaatg
     1621 ttataatgaa tatatagaat taccttagct ttaaaataga tttctaccca tttgataata
     1681 atctggaaga aaggaacaaa tatagttaaa gctatatagg tgaaagatat gccggctttt
     1741 gaaagggtta gatcaggtaa atggcattga aaccggatgg aaaccaggat gtgtaaagct
     1801 ttaaaaaatc ggcggatatt agacaggtta aaaaaaacga agtcattgca ttagactgaa
     1861 catatccaaa ttacaattga catatcaaat tataattgtt gaaaagctcc tgctaatatg
     1921 ttatcatatc actcggtcac gctgctaaaa aaatgtcgat atatggggat acggctaaac
     1981 ggagtatctc aaggcagttt gacaaatatt ttgcgggagt tataaaaatg gtcgatgttc
     2041 aaagccttac ggagcaaata cgaattttag tcaatcaaat ttctgtttta gtgataatgg
     2101 gactcatagg gtttattgca ggtaaaaccc gttatcttcc ggataattcg ggatcagtgc
     2161 tttcaagaac ggtaataaag cttacggcac cgctcctgat ttttacgacc ctggcgggac
     2221 gaaattttac taaagacatt ctgataaacg gaatgtggat attccttttg gcggtagtat
     2281 ttataatgtt ttcctttgta gtggggcatt ttgccggaaa gatattaaaa cttgaagggg
     2341 caactgcgaa cgtatttaaa atgcatttca tgtttggaaa cgtaatttat ctggcttttc
     2401 cccttttatc gtgggaatag gcatggaaaa gatttttttg cccgaggatg cattgataaa
     2461 gtatttcttt tccgaaagag aggaaaagat tctaaagagt tttggaaata ctgatgaata
     2521 ttgtgtgcag agtacaattc tatggacaag aaaagaggct ttgtcaaaac tttttcgtct
     2581 gggaatgagg atggatttta aaaagctgga tactttggag ggcgaggtgg tttttcagga
     2641 aacaaacagg gcgcgtctgt tttcttttat atgcaataat tactgtatct ctctggcatt
     2701 gccaggtttt aataaagatt aaaattattg actagaaata aaaaaattgt ccataatatt
     2761 aatggacaaa aaaacaaaga attacatcaa aggaagataa aaatactttg ttaaaaaatt
     2821 aattattttt tatctaaact attgaaaatg aaaataaaat aatataaaat gaatcatagt
     2881 gcaagagata cttgccagag gatgaatatt ttactgcatt catgctttat ggcagctaat
     2941 agaggcatta aattaaattt taatttacaa taggaggcga tattaatgaa ctttaataaa
     3001 attgatttag acaattggaa gagaaaagag atatttaatc attatttgaa ccaacaaacg
     3061 acttttagta taaccacaga aattgatatt agtgttttat accgaaacat aaaacaagaa
     3121 ggatataaat tttaccctgc atttattttc ttagtgacaa gggtgataaa ctcaaataca
     3181 gcttttagaa ctggttacaa tagcgacgga gagttaggtt attgggataa gttagagcca
     3241 ctttatacaa tttttgatgg tgtatctaaa acattctctg gtatttggac tcctgtaaag
     3301 aatgacttca aagagtttta tgatttatac ctttctgatg tagagaaata taatggttcg
     3361 gggaaattgt ttcccaaaac acctatacct gaaaatgctt tttctctttc tattattcca
     3421 tggacttcat ttactgggtt taacttaaat atcaataata atagtaatta ccttctaccc
     3481 attattacag caggaaaatt cattaataaa ggtaattcaa tatatttacc gctatcttta
     3541 caggtacatc attctgtttg tgatggttat catgcaggat tgtttatgaa ctctattcag
     3601 gaattgtcag ataggcctaa tgactggctt ttataataaa ggaggtcgac gtcatggaaa
     3661 atttatcaaa agacatcgat gaaattttga tcacagaaga agaacttaag gaaaagataa
     3721 aagagcttgg gaggcaaatc acaaaagact acaaagggaa aaatttgatg ttggtaggag
     3781 ttttaaaagg tgctttaatg tttatggctg atttgtcaag acacatagat ttgcctttat
     3841 cacttgattt tatggctgtt tccagctatg gaagctcaac tcattcatca ggaatagtaa
     3901 agataatcaa agatcttgat ataagcatag aaggcaaaga tgttctgatt gtggaagaca
     3961 taattgacag cggtttgact ttgtcttact taagggaaac tttacttgga aggaagccaa
     4021 aaagcctgaa aatatgcaca atattagaca aaccggagag aagagaagca tctgtaaaag
     4081 tcgattatgt aggatttaag atacctgata agtttgtcgt gggttatgga ttggactttg
     4141 atgaaaagta caggaacctt ccttttatag gcgttttgaa acctgaaatg tacagctaag
     4201 cccgggcctc gagaaaacaa aaggctcagt cggaagactg ggccttttgt tttggtaccg
     4261 aattcggcgc gcctcagcgt ttaaacccgc tgatcctaga gggccgcatc atgtaattag
     4321 ttatgtcacg cttacattca cgccctcccc atgctttaat acatacctgg cccagtagtt
     4381 cagtcctatc attgtgcgtt cgagcagttt cctggaagac ggatttggaa atgcttttgc
     4441 gggaaaatac gagtcgattt tctgcgtaaa cgcatgcgat ccggaagaaa ggcttttgca
     4501 atttgaagac gcggtgcgca gggtatatgc aagcacgatg gacaggtctg ctttggagta
     4561 ccgcagacaa cggggacttg ataaaatgga cgagcaaatg gcgattttgg tgcagcgtgt
     4621 cagcggaacg aaatttgaca aatattatat gccatgtgct gcgggagttg gtttttccta
     4681 tagtgtatac cgctggagtg atgaatttag tgcagatgcg ggacttctgc gtttggttgc
     4741 aggtctgggt acaaaggcgg tggacagaac gggtgtcgat tatccccggc tggtaaacct
     4801 ggataatcct gaaagtacca ttttaacgag gtcgagtgat aaacatcgtt tttcacagcg
     4861 aagagtggat gttattgatt tacagaaaaa tgaagtaaga gatattgatg taagtgaact
     4921 gattcctgag cttccggatt ggtatgtgaa tcttgtctgt gagcatgatt atgatactga
     4981 gagaatgttt tatgaaagag gacagagacg aaactgctaa tagtagtgaa aaaatcaggg
     5041 ataatgccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag gtccctattt
     5101 atttttttat agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt
     5161 ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt gagaaggttt
     5221 tgggacgctc gaaggcttta atttgcaagc tgcggccctg cattaatgaa tcggccaacg
     5281 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5341 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5401 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5461 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5521 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5581 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5641 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5701 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5761 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5821 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5881 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5941 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6001 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6061 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6121 gtggaacgaa aactcacgtt aagggatttt ggtcatgaaa aaatccgctt aagtccgcgg
     6181 gacttcatgg cactttctac accttgcttc ataagactcc tttgctccaa ccattataat
     6241 gggatcgtcg taattggcag gcttcccatt tataagcctt tgcgtgcgag tagcaggatt
     6301 gccacacttc atgcatatag cagtaagctt atcgacaaat tcagctatgg ccatcaattc
     6361 tggagtcgga ccaaatggtt cacctctaaa gtccatgtca agccctgcac atatgactct
     6421 tttcccgcta tcggcaatct cttttacgat gtcgactatt tcagaatcaa aaaattgaac
     6481 ttcatctata gcaaatacat ccgtatcttc ttcagcataa gccaatatgt cagaagcctt
     6541 tactatggca atggcgtgca tgttgtcgcc gttatgagat acgactttgt ctatggaata
     6601 cctatcgtct atagccggtt tgaaaacctg aactttttgc ctggcaatct tagctctctt
     6661 tatccttctt ataagttcct cactttttcc actgaacatg ggaccagtta caacttctat
     6721 gtacccgtgg tctttaggcc catacatctt tcgtcctcct taaaattttc gtttatttgt
     6781 tttttattta gtgtgcctaa aagcaccatc ttatacattc gtaccttata tctcttatct
     6841 tgtatctctt acctcatatc tcttattatt aactaccgaa gtatattttc ataacggaat
     6901 cttatgactt atttttctga ctgccgtttt accatacggt taatttatac ggaaacagct
     6961 aatctaacct tgctgaaggc atgtttacca aaatattata tctactttac catattaatt
     7021 caagtggtca ttttgatatt ttacgccaaa atttttagcg ttcctttact tgataattta
     7081 tccaaaaacc ttcattcgtc aaaacaatta tgatataatt aatttatgat atgattaatt
     7141 aatatttacg gataactaat catttcaatg caatagatca acatgcaaat caatatccac
     7201 cttactccga acattttgca ccaacgaatg cccagcacta tatcccggca agcaatataa
     7261 cgtctataat tttttatgat aacgataaaa ttaaatttta tgaaaggggt aaaagcatga
     7321 gaaaaatagc tgtattgtta attactttga cgttcttagt cacgactcga atttaggagg
     7381 cttacttgtc tgctttcttc attagaatca atcctttttt aaaagtcaat cccgtttgtt
     7441 gaactactct ttaataaaat aatttttccg ttcccaattc cacattgcaa taatagaaaa
     7501 tccatcttca tcggcttttt cgtcatcatc tgtatgaatc aaatcgcctt cttctgtgtc
     7561 atcaaggttt aattttttat gtatttcttt taacaaacca ccataggaga ttaacctttt
     7621 acggtgtaaa ccttcctcca aatcagacaa acgtttcaaa ttcttttctt catcatcggt
     7681 cataaaatcc gtatccttta caggatattt tgcagtttcg tcaattgccg attgtatatc
     7741 cgatttatat ttatttttcg gtcgaatcat ttgaactttt acatttggat catagtctaa
     7801 tttcattgcc tttttccaaa attgaatcca ttgtttttga ttcacgtagt tttctgtatt
     7861 cttaaaataa gttggttcca cacataccaa tacatgcatg tgctgattat aagaattatc
     7921 tttattattt attgtcactt ccgttgcacg cataaaacca acaagatttt tattaatttt
     7981 tttatattgc atcattcggc gaaatccttg agccatatct gacaaactct tatttaattc
     8041 ttcgccatca taaacatttt taactgttaa tgtgagaaac aaccaacgaa ctgttggctt
     8101 ttgtttaata acttcagcaa caaccttttg tgactgaatg ccatgtttca ttgctctcct
     8161 ccagttgcac attggacaaa gcctggattt acaaaaccac actcgataca actttctttc
     8221 gcctgtttca cgattttgtt tatactctaa tatttcagca caatctttta ctctttcagc
     8281 ctttttaaat tcaagaatat gcagaagttc aaagtaatca acattagcga ttttcttttc
     8341 tctccatggt ctcacttttc cactttttgt cttgtccact aaaacccttg atttttcatc
     8401 tgaataaatg ctactattag gacacataat attaaaagaa acccccatct atttagttat
     8461 ttgtttggtc acttataact ttaacagatg gggtttttct gtgcaaccaa ttttaagggt
     8521 tttcaatact ttaaaacaca tacataccaa cacttcaacg cacctttcag caactaaaat
     8581 aaaaatgacg ttatttctat atgtatcaag aatagaaaga actcgttttt cgctacgctc
     8641 aaaacgcaaa aaaagcactc attcgagtgc tttttcttat cgctccaaat catgcgattt
     8701 tttcctcttt gcttttcttt gctcacgaag ttctcgatca cgctgcaaaa catcttgaag
     8761 cgaaaaagta ttcttctttt cttccgatcg ctcatgctga cgcacgaaaa gccctctagg
     8821 cgcataggaa caactcctaa atgcatgtga ggggttttct cgtccatgtg aacagtcgca
     8881 tacgcaatat tttgtttccc atagcggccg cgattatcaa aaaggatctt cacctagatc
     8941 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     9001 gacag
//
