LOCUS       V007996                 8727 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V007996
VERSION     V007996
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8727)
  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 8727)
  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 8727)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8727)
  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..8727
                     /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="AYA22194.1"
                     /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..5332
                     /label="upstream homology recombination flank"
                     /note="upstream homology recombination flank"
     misc_feature    5333..5948
                     /label="downstream homology recombination flank"
                     /note="downstream homology recombination flank"
     regulatory      5952..6526
                     /label="C. thermocellum gapDH promoter"
                     /note="C. thermocellum gapDH promoter"
                     /regulatory_class="promoter"
     CDS             6527..7174
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     CDS             7195..7740
                     /codon_start=1
                     /gene="hpt"
                     /product="hypoxanthine phosphoribosyltransferase"
                     /label="hpt"
                     /protein_id="AYA22198.1"
                     /translation="MENLSKDIDEILITEEELKEKIKELGRQITKDYKGKNLMLVGVLK
                     GALMFMADLSRHIDLPLSLDFMAVSSYGSSTHSSGIVKIIKDLDISIEGKDVLIVEDII
                     DSGLTLSYLRETLLGRKPKSLKICTILDKPERREASVKVDYVGFKIPDKFVVGYGLDFD
                     EKYRNLPFIGVLKPEMYS"
     gene            7195..7740
                     /gene="hpt"
                     /label="hpt"
     misc_feature    7925..8484
                     /label="gene target internal homology recombination flank"
                     /note="gene target internal homology recombination flank"
     terminator      8517..8705
                     /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 ttttgcaatt
     4741 accgtgctgt ctgaaaagac ggaaccttca attataggaa cttttggatt ccagacaggc
     4801 aaagaaatta acaagtttga ttctgtaaag tatgatttaa aaaacgatat gccaattgta
     4861 aaggatgggt gtgcttatct tgtctgcgaa gtgattaaca agatggaaac aagcacccat
     4921 actgtttttt tgggaaaagt actggatgca gattttattt ccgacgaaga acccatgact
     4981 tattcctatt atcataaggt aataaaagga aaaagtccca agaatgctcc gacatatatc
     5041 ggagtggaga atgaaacaga ggaaaacaaa ctaaaggaaa aaagagcaaa atatgtgtgc
     5101 agtatatgtg gatatgaata tacaggggat ataccttttg aggacttgcc ggaagattat
     5161 gtctgtcctg tatgcggaca gcctaaatcg gtatttgtaa agaaagaata aggcataata
     5221 ttttatgggt gttttaatca aaatgaattt gggaaataat caaatattca tatgtaattt
     5281 caaaaatata aaccgaaaac gtaacgggaa agaataggag agtgaagaaa gttcgtaaca
     5341 ggtagcatgc aaaagggctg ctgcactaag aaattggtgc aacagccctt tttgtcagtc
     5401 ttgtgtcagg acggtgtttg ccttgggatt tgttcttatt tttgatttga atcggcatat
     5461 tccttggcgt tttccacttg aatttcactg ggaatgggaa cccctgactc gggttttttc
     5521 ttttcgatgt ttgcccaggc cgctgtgttg tgcttttcaa ttggtactgc catgtttttt
     5581 tgtttataat tgttttttga catcttatca ctccaaacaa gacttttata ttgttgtgtt
     5641 gatgttattt tatatcacgg aagatttatt atactgttca ttatttacag gtattatttg
     5701 ggagtttgct gcggtaaatt tgttttttta ctttttttcc ggtgtgagat ttcttctctt
     5761 ttttatagtt ttattttcat ttattttgac ttttttgcct gacttaattt ctggtattgt
     5821 tatagttttt ttagaaaaaa agattaaaga attgaatgta cagtttacat atgataaaat
     5881 tggtattatt ttatttaaat aaataatata taaaaggaga gaaatgagat gagttttttt
     5941 agggggagat cgtgggaata ggcatggaaa agattttttt gcccgaggat gcattgataa
     6001 agtatttctt ttccgaaaga gaggaaaaga ttctaaagag ttttggaaat actgatgaat
     6061 attgtgtgca gagtacaatt ctatggacaa gaaaagaggc tttgtcaaaa ctttttcgtc
     6121 tgggaatgag gatggatttt aaaaagctgg atactttgga gggcgaggtg gtttttcagg
     6181 aaacaaacag ggcgcgtctg ttttctttta tatgcaataa ttactgtatc tctctggcat
     6241 tgccaggttt taataaagat taaaattatt gactagaaat aaaaaaattg tccataatat
     6301 taatggacaa aaaaacaaag aattacatca aaggaagata aaaatacttt gttaaaaaat
     6361 taattatttt ttatctaaac tattgaaaat gaaaataaaa taatataaaa tgaatcatag
     6421 tgcaagagat acttgccaga ggatgaatat tttactgcat tcatgcttta tggcagctaa
     6481 tagaggcatt aaattaaatt ttaatttaca ataggaggcg atattaatga actttaataa
     6541 aattgattta gacaattgga agagaaaaga gatatttaat cattatttga accaacaaac
     6601 gacttttagt ataaccacag aaattgatat tagtgtttta taccgaaaca taaaacaaga
     6661 aggatataaa ttttaccctg catttatttt cttagtgaca agggtgataa actcaaatac
     6721 agcttttaga actggttaca atagcgacgg agagttaggt tattgggata agttagagcc
     6781 actttataca atttttgatg gtgtatctaa aacattctct ggtatttgga ctcctgtaaa
     6841 gaatgacttc aaagagtttt atgatttata cctttctgat gtagagaaat ataatggttc
     6901 ggggaaattg tttcccaaaa cacctatacc tgaaaatgct ttttctcttt ctattattcc
     6961 atggacttca tttactgggt ttaacttaaa tatcaataat aatagtaatt accttctacc
     7021 cattattaca gcaggaaaat tcattaataa aggtaattca atatatttac cgctatcttt
     7081 acaggtacat cattctgttt gtgatggtta tcatgcagga ttgtttatga actctattca
     7141 ggaattgtca gataggccta atgactggct tttataataa aggaggtcga cgtcatggaa
     7201 aatttatcaa aagacatcga tgaaattttg atcacagaag aagaacttaa ggaaaagata
     7261 aaagagcttg ggaggcaaat cacaaaagac tacaaaggga aaaatttgat gttggtagga
     7321 gttttaaaag gtgctttaat gtttatggct gatttgtcaa gacacataga tttgccttta
     7381 tcacttgatt ttatggctgt ttccagctat ggaagctcaa ctcattcatc aggaatagta
     7441 aagataatca aagatcttga tataagcata gaaggcaaag atgttctgat tgtggaagac
     7501 ataattgaca gcggtttgac tttgtcttac ttaagggaaa ctttacttgg aaggaagcca
     7561 aaaagcctga aaatatgcac aatattagac aaaccggaga gaagagaagc atctgtaaaa
     7621 gtcgattatg taggatttaa gatacctgat aagtttgtcg tgggttatgg attggacttt
     7681 gatgaaaagt acaggaacct tccttttata ggcgttttga aacctgaaat gtacagctaa
     7741 gcccgggcct cgagaaaaca aaaggctcag tcggaagact gggccttttg ttttggtacc
     7801 gaattcggcg cgcctcagcg tttaaacccg ctgatcctag agggccgcat catgtaatta
     7861 gttatgtcac gcttacattc acgccctccc catgctttaa tacatacctg gcccagtagt
     7921 tcagcaggga agagaagtac agaataaatt ctatgacaac gttcccgata ttgttgcagg
     7981 ctatatgcag gaaataaaaa ggattacagg ccgcgaatac cgcccgtttg attattacgg
     8041 ggcaaaagat gcggaatatg tgattgtggc aatgggttcc gtttgcgata ccattgatga
     8101 aaccattgac taccttatgg cgaagggaga aaaggtaggt tctgtaagag tacaccttta
     8161 caggcctttc tccgaaaagt acttctttga cgtaattcca aaaacggtga agaaaattgc
     8221 agtgcttgac aggactaaag aacccggagc accgggagag cctttgtacc ttgatgtttc
     8281 aaaaatgtat ctgtcaaaag aaaacaggcc ggttatcgta ggcggaagat atggactcgg
     8341 ttcgaaggat accagaccgt cccaaatcat atcagtattt gagaatttga agcaggatga
     8401 gccgaaaaac aactttacaa ttggaattgt cgatgatgtg accaatttgt ctcttccgga
     8461 aggggacata attgagacca caccctgcta atagtagtga aaaaatcagg gataatgccg
     8521 ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt tattttttta
     8581 tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtaca
     8641 gacgcgtgta cgcatgtaac attatactga aaaccttgct tgagaaggtt ttgggacgct
     8701 cgaaggcttt aatttgcaag ctgcggc
//
