LOCUS       V007999                 6950 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V007999
VERSION     V007999
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6950)
  AUTHORS   Hon S, Olson DG, Holwerda EK, Lanahan AA, Murphy SJL., Maloney MI,
            Zheng T, Papanek B, Guss AM, Lynd LR.
  TITLE     The ethanol pathway from Thermoanaerobacterium saccharolyticum
            improves ethanol production in Clostridium thermocellum
  JOURNAL   Metab. Eng. 42, 175-184 (2017)
   PUBMED   28663138
REFERENCE   2  (bases 1 to 6950)
  AUTHORS   Hon S, Olson DG, Holwerda EK, Lanahan AA, Murphy SJL., Maloney MI,
            Zheng T, Papanek BA, Guss AM, Lynd LR.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-MAR-2017) Thayer School of Engineering, Dartmouth
            College, 14 Engineering Drive, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 6950)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6950)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "Metab. Eng.
            42, 175-184 (2017)"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (28-MAR-2017) Thayer School of Engineering, Dartmouth College, 14
            Engineering Drive, Hanover, NH 03755, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6950
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1..579)
                     /codon_start=1
                     /gene="tdk"
                     /product="Tdk"
                     /label="tdk"
                     /note="thymidine kinase"
                     /protein_id="ASK05960.1"
                     /translation="MYGPKDHGYIEVVTGPMFSGKSEELIRRIKRAKIARQKVQVFKPA
                     IDDRYSIDKVVSHNGDNMHAIAIVKASDILAYAEEDTDVFAIDEVQFFDSEIVDIVKEI
                     ADSGKRVICAGLDMDFRGEPFGPTPELMAIAEFVDKLTAICMKCGNPATRTQRLINGKP
                     ANYDDPIIMVGAKESYEARCRKCHEVPRT"
     gene            complement(1..579)
                     /gene="tdk"
                     /label="tdk"
     regulatory      complement(580..1200)
                     /label="C therm cbp promoter"
                     /note="C therm cbp promoter"
                     /regulatory_class="promoter"
     CDS             complement(1279..2280)
                     /label="repB"
                     /note="RepB replication protein"
     rep_origin      complement(2281..2735)
                     /direction=LEFT
                     /label="C thermocellum replication origin"
                     /note="C thermocellum replication origin"
     CDS             complement(2841..3698)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(3699..3803)
                     /label="AmpR promoter"
     regulatory      3896..4470
                     /label="C therm gapDH promoter"
                     /note="C therm gapDH promoter"
                     /regulatory_class="promoter"
     CDS             4471..5118
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     CDS             5139..5684
                     /codon_start=1
                     /gene="hpt"
                     /product="Hpt"
                     /label="hpt"
                     /note="hypoxanthine phosphoribosyltransferase"
                     /protein_id="ASK05964.1"
                     /translation="MENLSKDIDEILITEEELKEKIKELGRQITKDYKGKNLMLVGVLK
                     GALMFMADLSRHIDLPLSLDFMAVSSYGSSTHSSGIVKIIKDLDISIEGKDVLIVEDII
                     DSGLTLSYLRETLLGRKPKSLKICTILDKPERREASVKVDYVGFKIPDKFVVGYGLDFD
                     EKYRNLPFIGVLKPEMYS"
     gene            5139..5684
                     /gene="hpt"
                     /label="hpt"
     terminator      5901..6089
                     /label="CYC1 terminator"
                     /note="transcription terminator for CYC1"
     rep_origin      6139..6684
                     /direction=RIGHT
                     /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."
ORIGIN
        1 ttaagtccgc gggacttcat ggcactttct acaccttgct tcataagact cctttgctcc
       61 aaccattata atgggatcgt cgtaattggc aggcttccca tttataagcc tttgcgtgcg
      121 agtagcagga ttgccacact tcatgcatat agcagtaagc ttatcgacaa attcagctat
      181 ggccatcaat tctggagtcg gaccaaatgg ttcacctcta aagtccatgt caagccctgc
      241 acatatgact cttttcccgc tatcggcaat ctcttttacg atgtcgacta tttcagaatc
      301 aaaaaattga acttcatcta tagcaaatac atccgtatct tcttcagcat aagccaatat
      361 gtcagaagcc tttactatgg caatggcgtg catgttgtcg ccgttatgag atacgacttt
      421 gtctatggaa tacctatcgt ctatagccgg tttgaaaacc tgaacttttt gcctggcaat
      481 cttagctctc tttatccttc ttataagttc ctcacttttt ccactgaaca tgggaccagt
      541 tacaacttct atgtacccgt ggtctttagg cccatacatc tttcgtcctc cttaaaattt
      601 tcgtttattt gttttttatt tagtgtgcct aaaagcacca tcttatacat tcgtacctta
      661 tatctcttat cttgtatctc ttacctcata tctcttatta ttaactaccg aagtatattt
      721 tcataacgga atcttatgac ttatttttct gactgccgtt ttaccatacg gttaatttat
      781 acggaaacag ctaatctaac cttgctgaag gcatgtttac caaaatatta tatctacttt
      841 accatattaa ttcaagtggt cattttgata ttttacgcca aaatttttag cgttccttta
      901 cttgataatt tatccaaaaa ccttcattcg tcaaaacaat tatgatataa ttaatttatg
      961 atatgattaa ttaatattta cggataacta atcatttcaa tgcaatagat caacatgcaa
     1021 atcaatatcc accttactcc gaacattttg caccaacgaa tgcccagcac tatatcccgg
     1081 caagcaatat aacgtctata attttttatg ataacgataa aattaaattt tatgaaaggg
     1141 gtaaaagcat gagaaaaata gctgtattgt taattacttt gacgttctta gtcacgactc
     1201 gaatttagga ggcttacttg tctgctttct tcattagaat caatcctttt ttaaaagtca
     1261 atcccgtttg ttgaactact ctttaataaa ataatttttc cgttcccaat tccacattgc
     1321 aataatagaa aatccatctt catcggcttt ttcgtcatca tctgtatgaa tcaaatcgcc
     1381 ttcttctgtg tcatcaaggt ttaatttttt atgtatttct tttaacaaac caccatagga
     1441 gattaacctt ttacggtgta aaccttcctc caaatcagac aaacgtttca aattcttttc
     1501 ttcatcatcg gtcataaaat ccgtatcctt tacaggatat tttgcagttt cgtcaattgc
     1561 cgattgtata tccgatttat atttattttt cggtcgaatc atttgaactt ttacatttgg
     1621 atcatagtct aatttcattg cctttttcca aaattgaatc cattgttttt gattcacgta
     1681 gttttctgta ttcttaaaat aagttggttc cacacatacc aatacatgca tgtgctgatt
     1741 ataagaatta tctttattat ttattgtcac ttccgttgca cgcataaaac caacaagatt
     1801 tttattaatt tttttatatt gcatcattcg gcgaaatcct tgagccatat ctgacaaact
     1861 cttatttaat tcttcgccat cataaacatt tttaactgtt aatgtgagaa acaaccaacg
     1921 aactgttggc ttttgtttaa taacttcagc aacaaccttt tgtgactgaa tgccatgttt
     1981 cattgctctc ctccagttgc acattggaca aagcctggat ttacaaaacc acactcgata
     2041 caactttctt tcgcctgttt cacgattttg tttatactct aatatttcag cacaatcttt
     2101 tactctttca gcctttttaa attcaagaat atgcagaagt tcaaagtaat caacattagc
     2161 gattttcttt tctctccatg gtctcacttt tccacttttt gtcttgtcca ctaaaaccct
     2221 tgatttttca tctgaataaa tgctactatt aggacacata atattaaaag aaacccccat
     2281 ctatttagtt atttgtttgg tcacttataa ctttaacaga tggggttttt ctgtgcaacc
     2341 aattttaagg gttttcaata ctttaaaaca catacatacc aacacttcaa cgcacctttc
     2401 agcaactaaa ataaaaatga cgttatttct atatgtatca agaatagaaa gaactcgttt
     2461 ttcgctacgc tcaaaacgca aaaaaagcac tcattcgagt gctttttctt atcgctccaa
     2521 atcatgcgat tttttcctct ttgcttttct ttgctcacga agttctcgat cacgctgcaa
     2581 aacatcttga agcgaaaaag tattcttctt ttcttccgat cgctcatgct gacgcacgaa
     2641 aagccctcta ggcgcatagg aacaactcct aaatgcatgt gaggggtttt ctcgtccatg
     2701 tgaacagtcg catacgcaat attttgtttc ccatagcggc cgcgattatc aaaaaggatc
     2761 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     2821 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     2881 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     2941 cgcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     3001 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     3061 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     3121 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     3181 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     3241 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     3301 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     3361 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     3421 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     3481 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     3541 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     3601 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     3661 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     3721 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     3781 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     3841 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcg atatcgtggg
     3901 aataggcatg gaaaagattt ttttgcccga ggatgcattg ataaagtatt tcttttccga
     3961 aagagaggaa aagattctaa agagttttgg aaatactgat gaatattgtg tgcagagtac
     4021 aattctatgg acaagaaaag aggctttgtc aaaacttttt cgtctgggaa tgaggatgga
     4081 ttttaaaaag ctggatactt tggagggcga ggtggttttt caggaaacaa acagggcgcg
     4141 tctgttttct tttatatgca ataattactg tatctctctg gcattgccag gttttaataa
     4201 agattaaaat tattgactag aaataaaaaa attgtccata atattaatgg acaaaaaaac
     4261 aaagaattac atcaaaggaa gataaaaata ctttgttaaa aaattaatta ttttttatct
     4321 aaactattga aaatgaaaat aaaataatat aaaatgaatc atagtgcaag agatacttgc
     4381 cagaggatga atattttact gcattcatgc tttatggcag ctaatagagg cattaaatta
     4441 aattttaatt tacaatagga ggcgatatta atgaacttta ataaaattga tttagacaat
     4501 tggaagagaa aagagatatt taatcattat ttgaaccaac aaacgacttt tagtataacc
     4561 acagaaattg atattagtgt tttataccga aacataaaac aagaaggata taaattttac
     4621 cctgcattta ttttcttagt gacaagggtg ataaactcaa atacagcttt tagaactggt
     4681 tacaatagcg acggagagtt aggttattgg gataagttag agccacttta tacaattttt
     4741 gatggtgtat ctaaaacatt ctctggtatt tggactcctg taaagaatga cttcaaagag
     4801 ttttatgatt tatacctttc tgatgtagag aaatataatg gttcggggaa attgtttccc
     4861 aaaacaccta tacctgaaaa tgctttttct ctttctatta ttccatggac ttcatttact
     4921 gggtttaact taaatatcaa taataatagt aattaccttc tacccattat tacagcagga
     4981 aaattcatta ataaaggtaa ttcaatatat ttaccgctat ctttacaggt acatcattct
     5041 gtttgtgatg gttatcatgc aggattgttt atgaactcta ttcaggaatt gtcagatagg
     5101 cctaatgact ggcttttata ataaaggagg tcgacgtcat ggaaaattta tcaaaagaca
     5161 tcgatgaaat tttgatcaca gaagaagaac ttaaggaaaa gataaaagag cttgggaggc
     5221 aaatcacaaa agactacaaa gggaaaaatt tgatgttggt aggagtttta aaaggtgctt
     5281 taatgtttat ggctgatttg tcaagacaca tagatttgcc tttatcactt gattttatgg
     5341 ctgtttccag ctatggaagc tcaactcatt catcaggaat agtaaagata atcaaagatc
     5401 ttgatataag catagaaggc aaagatgttc tgattgtgga agacataatt gacagcggtt
     5461 tgactttgtc ttacttaagg gaaactttac ttggaaggaa gccaaaaagc ctgaaaatat
     5521 gcacaatatt agacaaaccg gagagaagag aagcatctgt aaaagtcgat tatgtaggat
     5581 ttaagatacc tgataagttt gtcgtgggtt atggattgga ctttgatgaa aagtacagga
     5641 accttccttt tataggcgtt ttgaaacctg aaatgtacag ctaagcccgg gcctcgagaa
     5701 aacaaaaggc tcagtcggaa gactgggcct tttgttttgg taccgaattc ggcgcgcctc
     5761 agcgtttaaa cccgctgatc ctagagggcc gcatcatgta attagttatg tcacgcttac
     5821 attcacgccc tccccatgct ttaatacata cctggcccag tagttcagct gctaatagta
     5881 gtgaaaaaat cagggataat gccgctctaa ccgaaaagga aggagttaga caacctgaag
     5941 tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta tttatatttc
     6001 aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata ctgaaaacct
     6061 tgcttgagaa ggttttggga cgctcgaagg ctttaatttg caagctgcgg cttttatctg
     6121 attaataaga tgatcttctt gagatcgttt tggtctgcgc gtaatctctt gctctgaaaa
     6181 cgaaaaaacc gccttgcagg gcggtttttc gaaggttctc tgagctacca actctttgaa
     6241 ccgaggtaac tggcttggag gagcgcagtc accaaaactt gtcctttcag tttagcctta
     6301 accggcgcat gacttcaaga ctaactcctc taaatcaatt accagtggct gctgccagtg
     6361 gtgcttttgc atgtctttcc gggttggact caagacgata gttaccggat aaggcgcagc
     6421 ggtcggactg aacggggggt tcgtgcatac agtccagctt ggagcgaact gcctacccgg
     6481 aactgagtgt caggcgtgga atgagacaaa cgcggccata acagcggaat gacaccggta
     6541 aaccgaaagg caggaacagg agagcgcacg agggagccgc cagggggaaa cgcctggtat
     6601 ctttatagtc ctgtcgggtt tcgccaccac tgatttgagc gtcagatttc gtgatgcttg
     6661 tcaggggggc ggagcctatg gaaaaacggc tttgccgcgg ccctctcact tccctgttaa
     6721 gtatcttcct ggcatcttcc aggaaatctc cgccccgttc gtaagccatt tccgctcgcc
     6781 gcagtcgaac gaccgagcgt agcgagtcag tgagcgagga agcggaatat atcctgtatc
     6841 acatattctg ctgacgcacc ggtgcagcct tttttctcct gccacatgaa gcacttcact
     6901 gacaccctca tcagtgccaa catagtaagc cagtatacaa aaaaatccgc
//
