LOCUS       V003238                 8436 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003238
VERSION     V003238
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8436)
  AUTHORS   Hon S, Holwerda EK, Worthen RS, Maloney MI, Tian L, Cui J, Lin PP,
            Lynd LR, Olson DG.
  TITLE     Expressing the Thermoanaerobacterium saccharolyticum pforA in
            engineered Clostridium thermocellum improves ethanol production
  JOURNAL   Biotechnol Biofuels 11, 242 (2018)
   PUBMED   30202437
REFERENCE   2  (bases 1 to 8436)
  AUTHORS   Hon S, Olson DG, Holwerda EK, Worthen RS, Maloney MI, Tian L, Cui J,
            Lin PP, Lynd LR.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-APR-2018) Thayer School of Engineering, Dartmouth
            College, 14 Engineering Drive, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 8436)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8436)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "Biotechnol
            Biofuels 11, 242 (2018)"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (20-APR-2018) Thayer School of Engineering, Dartmouth College, 14
            Engineering Drive, Hanover, NH 03755, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8436
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      28..573
                     /label="p15A ori"
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells
                     that contain a second plasmid with the ColE1 origin."
     CDS             complement(840..1418)
                     /codon_start=1
                     /gene="tdk"
                     /product="thymidine kinase"
                     /label="tdk"
                     /protein_id="AYA22147.1"
                     /translation="MYGPKDHGYIEVVTGPMFSGKSEELIRRIKRAKIARQKVQVFKPA
                     IDDRYSIDKVVSHNGDNMHAIAIVKASDILAYAEEDTDVFAIDEVQFFDSEIVDIVKEI
                     ADSGKRVICAGLDMDFRGEPFGPTPELMAIAEFVDKLTAICMKCGNPATRTQRLINGKP
                     ANYDDPIIMVGAKESYEARCRKCHEVPRT"
     gene            complement(840..1418)
                     /gene="tdk"
                     /label="tdk"
     regulatory      complement(1419..2039)
                     /label="C. thermocellum cbp promoter"
                     /note="C. thermocellum cbp promoter"
                     /regulatory_class="promoter"
     CDS             complement(2118..3119)
                     /label="repB"
                     /note="RepB replication protein"
     rep_origin      complement(3120..3574)
                     /direction=LEFT
                     /label="C. thermocellum origin of replication"
                     /note="C. thermocellum origin of replication"
     CDS             complement(3680..4537)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(4538..4642)
                     /label="AmpR promoter"
     misc_feature    4732..5234
                     /label="upstream homology recombination flank"
                     /note="upstream homology recombination flank"
     misc_feature    5235..5733
                     /label="downstream homology recombination flank"
                     /note="downstream homology recombination flank"
     regulatory      5737..6311
                     /label="C. thermocellum gapDH promoter"
                     /note="C. thermocellum gapDH promoter"
                     /regulatory_class="promoter"
     CDS             6312..6959
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     CDS             6980..7525
                     /codon_start=1
                     /gene="hpt"
                     /product="hypoxanthine phosphoribosyltransferase"
                     /label="hpt"
                     /protein_id="AYA22151.1"
                     /translation="MENLSKDIDEILITEEELKEKIKELGRQITKDYKGKNLMLVGVLK
                     GALMFMADLSRHIDLPLSLDFMAVSSYGSSTHSSGIVKIIKDLDISIEGKDVLIVEDII
                     DSGLTLSYLRETLLGRKPKSLKICTILDKPERREASVKVDYVGFKIPDKFVVGYGLDFD
                     EKYRNLPFIGVLKPEMYS"
     gene            6980..7525
                     /gene="hpt"
                     /label="hpt"
     CDS             complement(8075..8086)
                     /label="Factor Xa site"
                     /note="Factor Xa recognition and cleavage site"
     terminator      8226..8414
                     /label="CYC1 terminator"
                     /note="transcription terminator for CYC1"
ORIGIN
        1 ttttatctga ttaataagat gatcttcttg agatcgtttt ggtctgcgcg taatctcttg
       61 ctctgaaaac gaaaaaaccg ccttgcaggg cggtttttcg aaggttctct gagctaccaa
      121 ctctttgaac cgaggtaact ggcttggagg agcgcagtca ccaaaacttg tcctttcagt
      181 ttagccttaa ccggcgcatg acttcaagac taactcctct aaatcaatta ccagtggctg
      241 ctgccagtgg tgcttttgca tgtctttccg ggttggactc aagacgatag ttaccggata
      301 aggcgcagcg gtcggactga acggggggtt cgtgcataca gtccagcttg gagcgaactg
      361 cctacccgga actgagtgtc aggcgtggaa tgagacaaac gcggccataa cagcggaatg
      421 acaccggtaa accgaaaggc aggaacagga gagcgcacga gggagccgcc agggggaaac
      481 gcctggtatc tttatagtcc tgtcgggttt cgccaccact gatttgagcg tcagatttcg
      541 tgatgcttgt caggggggcg gagcctatgg aaaaacggct ttgccgcggc cctctcactt
      601 ccctgttaag tatcttcctg gcatcttcca ggaaatctcc gccccgttcg taagccattt
      661 ccgctcgccg cagtcgaacg accgagcgta gcgagtcagt gagcgaggaa gcggaatata
      721 tcctgtatca catattctgc tgacgcaccg gtgcagcctt ttttctcctg ccacatgaag
      781 cacttcactg acaccctcat cagtgccaac atagtaagcc agtatacaaa aaaatccgct
      841 taagtccgcg ggacttcatg gcactttcta caccttgctt cataagactc ctttgctcca
      901 accattataa tgggatcgtc gtaattggca ggcttcccat ttataagcct ttgcgtgcga
      961 gtagcaggat tgccacactt catgcatata gcagtaagct tatcgacaaa ttcagctatg
     1021 gccatcaatt ctggagtcgg accaaatggt tcacctctaa agtccatgtc aagccctgca
     1081 catatgactc ttttcccgct atcggcaatc tcttttacga tgtcgactat ttcagaatca
     1141 aaaaattgaa cttcatctat agcaaataca tccgtatctt cttcagcata agccaatatg
     1201 tcagaagcct ttactatggc aatggcgtgc atgttgtcgc cgttatgaga tacgactttg
     1261 tctatggaat acctatcgtc tatagccggt ttgaaaacct gaactttttg cctggcaatc
     1321 ttagctctct ttatccttct tataagttcc tcactttttc cactgaacat gggaccagtt
     1381 acaacttcta tgtacccgtg gtctttaggc ccatacatct ttcgtcctcc ttaaaatttt
     1441 cgtttatttg ttttttattt agtgtgccta aaagcaccat cttatacatt cgtaccttat
     1501 atctcttatc ttgtatctct tacctcatat ctcttattat taactaccga agtatatttt
     1561 cataacggaa tcttatgact tatttttctg actgccgttt taccatacgg ttaatttata
     1621 cggaaacagc taatctaacc ttgctgaagg catgtttacc aaaatattat atctacttta
     1681 ccatattaat tcaagtggtc attttgatat tttacgccaa aatttttagc gttcctttac
     1741 ttgataattt atccaaaaac cttcattcgt caaaacaatt atgatataat taatttatga
     1801 tatgattaat taatatttac ggataactaa tcatttcaat gcaatagatc aacatgcaaa
     1861 tcaatatcca ccttactccg aacattttgc accaacgaat gcccagcact atatcccggc
     1921 aagcaatata acgtctataa ttttttatga taacgataaa attaaatttt atgaaagggg
     1981 taaaagcatg agaaaaatag ctgtattgtt aattactttg acgttcttag tcacgactcg
     2041 aatttaggag gcttacttgt ctgctttctt cattagaatc aatccttttt taaaagtcaa
     2101 tcccgtttgt tgaactactc tttaataaaa taatttttcc gttcccaatt ccacattgca
     2161 ataatagaaa atccatcttc atcggctttt tcgtcatcat ctgtatgaat caaatcgcct
     2221 tcttctgtgt catcaaggtt taatttttta tgtatttctt ttaacaaacc accataggag
     2281 attaaccttt tacggtgtaa accttcctcc aaatcagaca aacgtttcaa attcttttct
     2341 tcatcatcgg tcataaaatc cgtatccttt acaggatatt ttgcagtttc gtcaattgcc
     2401 gattgtatat ccgatttata tttatttttc ggtcgaatca tttgaacttt tacatttgga
     2461 tcatagtcta atttcattgc ctttttccaa aattgaatcc attgtttttg attcacgtag
     2521 ttttctgtat tcttaaaata agttggttcc acacatacca atacatgcat gtgctgatta
     2581 taagaattat ctttattatt tattgtcact tccgttgcac gcataaaacc aacaagattt
     2641 ttattaattt ttttatattg catcattcgg cgaaatcctt gagccatatc tgacaaactc
     2701 ttatttaatt cttcgccatc ataaacattt ttaactgtta atgtgagaaa caaccaacga
     2761 actgttggct tttgtttaat aacttcagca acaacctttt gtgactgaat gccatgtttc
     2821 attgctctcc tccagttgca cattggacaa agcctggatt tacaaaacca cactcgatac
     2881 aactttcttt cgcctgtttc acgattttgt ttatactcta atatttcagc acaatctttt
     2941 actctttcag cctttttaaa ttcaagaata tgcagaagtt caaagtaatc aacattagcg
     3001 attttctttt ctctccatgg tctcactttt ccactttttg tcttgtccac taaaaccctt
     3061 gatttttcat ctgaataaat gctactatta ggacacataa tattaaaaga aacccccatc
     3121 tatttagtta tttgtttggt cacttataac tttaacagat ggggtttttc tgtgcaacca
     3181 attttaaggg ttttcaatac tttaaaacac atacatacca acacttcaac gcacctttca
     3241 gcaactaaaa taaaaatgac gttatttcta tatgtatcaa gaatagaaag aactcgtttt
     3301 tcgctacgct caaaacgcaa aaaaagcact cattcgagtg ctttttctta tcgctccaaa
     3361 tcatgcgatt ttttcctctt tgcttttctt tgctcacgaa gttctcgatc acgctgcaaa
     3421 acatcttgaa gcgaaaaagt attcttcttt tcttccgatc gctcatgctg acgcacgaaa
     3481 agccctctag gcgcatagga acaactccta aatgcatgtg aggggttttc tcgtccatgt
     3541 gaacagtcgc atacgcaata ttttgtttcc catagcggcc gcgattatca aaaaggatct
     3601 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3661 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     3721 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagc
     3781 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     3841 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     3901 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     3961 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4021 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4081 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     4141 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4201 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     4261 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     4321 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     4381 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4441 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     4501 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     4561 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     4621 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     4681 ccattattat catgacatta acctataaaa ataggcgtat cacgaggcga tccatatctt
     4741 gcagttccct gtcaaaaact tattccatca ccggatggag actggggtat ttgattgcac
     4801 cctcttacat tgttgacggg gccaggaagg ttcatgactt tctgacagtg ggcgctgccg
     4861 caccgttgca ggaagcggca gtggttgccc tgaatttcgg ggatgattat tatgagaact
     4921 taaaaagaat ttatacagag aaaagggatt tttttctgga tggtttggat aggctggggc
     4981 ttgcgtatac cgtaccgcaa ggggcctatt atgtaatggt ggacatttcg gagtttggag
     5041 caaaaagcga cttggaattt tgtgagtgga tggcgaggga agtgggcgtt gcagcggttc
     5101 ccggctcaag ctttttcaga gataatgtaa atcatctgat tcgcttccac tttgcaaaaa
     5161 agaaagaaac tctggctgaa gctgtaaaga gacttgaaaa gcttaaagac aaagcaaggg
     5221 agcgatggag atgatttgcg tgactataga acagcatgta taaacattac agggaatatg
     5281 gacattatag ggaacatgga ctttttttga aaaaaggagg ggttgctcat gacaaaaggc
     5341 gctgaagaac tgtttcggga aatagagcta aaaaatatta ttctaagaga acagtgcaat
     5401 aattgccgcc taagcaggca tgagctggag aaaaccgaaa gggagctgga tgcccttctt
     5461 tacaaatact ataaaatata caaatgcaat ctttgcaaag tttgtgaaga aaagtgtccg
     5521 gaggagccgg aagaacaata attaattcaa aattcaaaaa ttatttgtta gcttttattg
     5581 acaagagagc atatatgata ctataatata aaacatattg gaatagtcgg gaaataaagg
     5641 cggtgattat aaaatgtaca ttgaggtcgt aaaaaccaat aaagctcctg aagcaatcgg
     5701 tccgtattcg caggcgattg ttacaggttc gttatcgtgg gaataggcat ggaaaagatt
     5761 tttttgcccg aggatgcatt gataaagtat ttcttttccg aaagagagga aaagattcta
     5821 aagagttttg gaaatactga tgaatattgt gtgcagagta caattctatg gacaagaaaa
     5881 gaggctttgt caaaactttt tcgtctggga atgaggatgg attttaaaaa gctggatact
     5941 ttggagggcg aggtggtttt tcaggaaaca aacagggcgc gtctgttttc ttttatatgc
     6001 aataattact gtatctctct ggcattgcca ggttttaata aagattaaaa ttattgacta
     6061 gaaataaaaa aattgtccat aatattaatg gacaaaaaaa caaagaatta catcaaagga
     6121 agataaaaat actttgttaa aaaattaatt attttttatc taaactattg aaaatgaaaa
     6181 taaaataata taaaatgaat catagtgcaa gagatacttg ccagaggatg aatattttac
     6241 tgcattcatg ctttatggca gctaatagag gcattaaatt aaattttaat ttacaatagg
     6301 aggcgatatt aatgaacttt aataaaattg atttagacaa ttggaagaga aaagagatat
     6361 ttaatcatta tttgaaccaa caaacgactt ttagtataac cacagaaatt gatattagtg
     6421 ttttataccg aaacataaaa caagaaggat ataaatttta ccctgcattt attttcttag
     6481 tgacaagggt gataaactca aatacagctt ttagaactgg ttacaatagc gacggagagt
     6541 taggttattg ggataagtta gagccacttt atacaatttt tgatggtgta tctaaaacat
     6601 tctctggtat ttggactcct gtaaagaatg acttcaaaga gttttatgat ttataccttt
     6661 ctgatgtaga gaaatataat ggttcgggga aattgtttcc caaaacacct atacctgaaa
     6721 atgctttttc tctttctatt attccatgga cttcatttac tgggtttaac ttaaatatca
     6781 ataataatag taattacctt ctacccatta ttacagcagg aaaattcatt aataaaggta
     6841 attcaatata tttaccgcta tctttacagg tacatcattc tgtttgtgat ggttatcatg
     6901 caggattgtt tatgaactct attcaggaat tgtcagatag gcctaatgac tggcttttat
     6961 aataaaggag gtcgacgtca tggaaaattt atcaaaagac atcgatgaaa ttttgatcac
     7021 agaagaagaa cttaaggaaa agataaaaga gcttgggagg caaatcacaa aagactacaa
     7081 agggaaaaat ttgatgttgg taggagtttt aaaaggtgct ttaatgttta tggctgattt
     7141 gtcaagacac atagatttgc ctttatcact tgattttatg gctgtttcca gctatggaag
     7201 ctcaactcat tcatcaggaa tagtaaagat aatcaaagat cttgatataa gcatagaagg
     7261 caaagatgtt ctgattgtgg aagacataat tgacagcggt ttgactttgt cttacttaag
     7321 ggaaacttta cttggaagga agccaaaaag cctgaaaata tgcacaatat tagacaaacc
     7381 ggagagaaga gaagcatctg taaaagtcga ttatgtagga tttaagatac ctgataagtt
     7441 tgtcgtgggt tatggattgg actttgatga aaagtacagg aaccttcctt ttataggcgt
     7501 tttgaaacct gaaatgtaca gctaagcccg ggcctcgaga aaacaaaagg ctcagtcgga
     7561 agactgggcc ttttgttttg gtaccgaatt cggcgcgcct cagcgtttaa acccgctgat
     7621 cctagagggc cgcatcatgt aattagttat gtcacgctta cattcacgcc ctccccatgc
     7681 tttaatacat acctggccca gtagttcagt tttaccagga ttgacggtaa gaagatttac
     7741 gaccgcagcg aaatcagaaa atgtgatttt attgtggttt tggatgaaac cctttttagt
     7801 gaaagctttt ttgatgattt gaaggaaaat ggaagaatca ttgtaaattc ggccgacaaa
     7861 gagttttacg caaaatatga ttcaaagagg ataactgtgg tggatgcgtc ctctgttgct
     7921 ctggaaatct taggcaaacc tataacaaat accgccatgc ttggcgccct ggttgcagtg
     7981 tcggatattg ttgacttgga agcagtagaa aaaggaatgg cagtttttct gaaaggggag
     8041 cttcttaaga aaaatattga agtggttcaa agaaccttcc ttcaatgcag ggaggcggtg
     8101 tcatgaacag gcccgaactt agagagtatg ttaaaccaaa gcatattaaa gattatcctg
     8161 tgggtccctg ctatacggca gggtatcttg taactgctaa tagtagtgaa aaaatcaggg
     8221 ataatgccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag gtccctattt
     8281 atttttttat agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt
     8341 ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt gagaaggttt
     8401 tgggacgctc gaaggcttta atttgcaagc tgcggc
//
