LOCUS       V007678                 9901 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V007678
VERSION     V007678
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9901)
  AUTHORS   Mearls EB, Olson DG, Herring CD, Lynd LR.
  TITLE     Development of a regulatable plasmid based gene expression system
            for Clostridium thermocellum and discovery of a histidine kinase
            involved in sporulation
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9901)
  AUTHORS   Argyros A, Mearls EB.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-MAR-2013) Engineering, Dartmouth College, 14
            Engineering Dr, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 9901)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9901)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (13-MAR-2013) Engineering, Dartmouth College, 14 Engineering Dr,
            Hanover, NH 03755, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9901
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      49..296
                     /label="CYC1 terminator"
                     /note="transcription terminator for CYC1"
     rep_origin      complement(544..1132)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(1306..2163)
                     /label="AmpR"
                     /note="beta-lactamase"
     CDS             complement(2262..3062)
                     /label="URA3"
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     promoter        complement(3063..3283)
                     /label="URA3 promoter"
     misc_feature    3311..3814
                     /label="CEN/ARS"
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     CDS             complement(4046..5047)
                     /label="repB"
                     /note="RepB replication protein"
     regulatory      5533..6180
                     /gene="gapD"
                     /regulatory_class="promoter"
     gene            5533..6180
                     /gene="gapD"
                     /label="gapD"
     CDS             6181..6828
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     regulatory      7808..7895
                     /regulatory_class="terminator"
     regulatory      7855..7893
                     /gene="celC"
                     /regulatory_class="promoter"
     gene            7855..7893
                     /gene="celC"
                     /label="celC"
     CDS             8023..8832
                     /codon_start=1
                     /gene="spo0A"
                     /product="Spo0A"
                     /label="spo0A"
                     /note="sporulation response regulator A"
                     /protein_id="AGT95724.1"
                     /translation="MTSNKIRVVIADDNREFGDILYEYLNNQEDIEVVGVARDGIEAYE
                     LIVEKLPDIAILDIIMPHLDGLGVLEKIGATAISKRPLFIILSAVGQDKITQRALALGA
                     EYYVVKPFDMEVLISRIRQLKNVNQPNVIRQDGLSGEVKSSYHPPQPKNLEAEVTNIMH
                     EIGVPAHIKGYQYLRDAIIMVVKDLDVINSITKQLYPTIAKEYNTTPSRVERAIRHAIE
                     VAWSRGQIDTIDSLFGYTINVGKGKPTNSEFIAMVADKLRLEMKMAT"
     gene            8023..8832
                     /gene="spo0A"
                     /label="spo0A"
     CDS             8859..9896
                     /codon_start=1
                     /gene="glyr3"
                     /product="GlyR3"
                     /label="glyr3"
                     /note="repressor"
                     /protein_id="AGT95725.1"
                     /translation="MTSEEIAKLCGVSRATVSRVINNSPNVKEETRQKILAVIKEKNYV
                     PIAPARRLAGIDSNIIGLFVLDIDISESKSRVSESTYFSRLINLIIDQANNFGFQVLVS
                     IITSQKQLSEIRNLFMSRTIFSGIFIGAFNDEIQLDDDIIMQHPTIIIDRQSERMVKKP
                     NRLVVNLDNFEGAYNATQFLIKLGHTRIGHISGDLRKLSGIERYEGYKKALEDAGLGFD
                     KNLVREGNFLDDSGYRLAREILKENVTAIFCANDVMAISAIKAIKETGLSVPDDISVIG
                     FDNTAIGNYIMPALTTVNAPLEHIAEACIESLKYFCEHKHFKQKEIRVKTDLIIRDSTK
                     RALEF"
     gene            8859..9896
                     /gene="glyr3"
                     /label="glyr3"
ORIGIN
        1 aattcggcgc gcctcagcgt ttaaacccgc tgatcctaga gggccgcatc atgtaattag
       61 ttatgtcacg cttacattca cgccctcccc ccacatccgc tctaaccgaa aaggaaggag
      121 ttagacaacc tgaagtctag gtccctattt atttttttat agttatgtta gtattaagaa
      181 cgttatttat atttcaaatt tttctttttt ttctgtacag acgcgtgtac gcatgtaaca
      241 ttatactgaa aaccttgctt gagaaggttt tgggacgctc gaaggcttta atttgcaagc
      301 tgcggccctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
      361 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
      421 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
      481 gaacatgtga gcaaaaggcc agcaaaagcc caggaaccgt aaaaaggccg cgttgctggc
      541 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
      601 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
      661 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
      721 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      781 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
      841 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
      901 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
      961 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     1021 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     1081 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     1141 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     1201 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     1261 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     1321 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     1381 cgtgtagata actacgatac gggagcgctt accatctggc cccagtgctg caatgatacc
     1441 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     1501 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     1561 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg gcattgctac
     1621 aggcatcgtg gtgtcactct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     1681 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     1741 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     1801 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     1861 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     1921 acgggataat agtgtatcac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     1981 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     2041 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     2101 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     2161 catactcttc ctttttcaat gggtaataac tgatataatt aaattgaagc tctaatttgt
     2221 gagtttagta tacatgcatt tacttataat acagtttttt agttttgctg gccgcatctt
     2281 ctcaaatatg cttcccagcc tgcttttctg taacgttcac cctctacctt agcatccctt
     2341 ccctttgcaa atagtcctct tccaacaata ataatgtcag atcctgtaga gaccacatca
     2401 tccacggttc tatactgttg acccaatgcg tctcccttgt catctaaacc cacaccgggt
     2461 gtcataatca accaatcgta accttcatct cttccaccca tgtctctttg agcaataaag
     2521 ccgataacaa aatctttgtc gctcttcgca atgtcaacag tacccttagt atattctcca
     2581 gtagataggg agcccttgca tgacaattct gctaacatca aaaggcctct aggttccttt
     2641 gttacttctt ctgccgcctg cttcaaaccg ctaacaatac ctgggcccac cacaccgtgt
     2701 gcattcgtaa tgtctgccca ttctgctatt ctgtatacac ccgcagagta ctgcaatttg
     2761 actgtattac caatgtcagc aaattttctg tcttcgaaga gtaaaaaatt gtacttggcg
     2821 gataatgcct ttagcggctt aactgtgccc tccatggaaa aatcagtcaa gatatccaca
     2881 tgtgttttta gtaaacaaat tttgggacct aatgcttcaa ctaactccag taattccttg
     2941 gtggtacgaa catccaatga agcacacaag tttgtttgct tttcgtgcat gatattaaat
     3001 agcttggcag caacaggact aggatgagta gcagcacgtt ccttatatgt agctttcgac
     3061 atgatttatc ttcgtttcct gcaggttttt gttctgtgca gttgggttaa gaatactggg
     3121 caatttcatg tttcttcaac actacatatg cgtatatata ccaatctaag tctgtgctcc
     3181 ttccttcgtt cttccttctg ttcggagatt accgaatcaa aaaaatttca aagaaaccga
     3241 aatcaaaaaa aagaataaaa aaaaaatgat gaattgaatt gaaaagctag cttatcgatg
     3301 ggtccttttc atcacgtgct ataaaaataa ttataattta aattttttaa tataaatata
     3361 taaattaaaa atagaaagta aaaaaagaaa ttaaagaaaa aatagttttt gttttccgaa
     3421 gatgtaaaag actctagggg gatcgccaac aaatactacc ttttatcttg ctcttcctgc
     3481 tctcaggtat taatgccgaa ttgtttcatc ttgtctgtgt agaagaccac acacgaaaat
     3541 cctgtgattt tacattttac ttatcgttaa tcgaatgtat atctatttaa tctgcttttc
     3601 ttgtctaata aatatatatg taaagtacgc tttttgttga aattttttaa acctttgttt
     3661 attttttttt cttcattccg taactcttct accttcttta tttactttct aaaatccaaa
     3721 tacaaaacat aaaaataaat aaacacagag taaattccca aattattcca tcattaaaag
     3781 atacgaggcg cgtgtaagtt acaggcaagc gatccgtccg ccggcgaacg tggcgagaaa
     3841 ggaagggaag aaagcgaaag gagcgggggc tagggcggtg ggaagtgtag gggtcacgct
     3901 gggcgtaacc accacacccg ccgcgcttaa tggggcgcta cagggcgcgt ggggatgatc
     3961 cactagtgaa tttaggaggc ttacttgtct gctttcttca ttagaatcaa tcctttttta
     4021 aaagtcaatc ccgtttgttg aactactctt taataaaata atttttccgt tcccaattcc
     4081 acattgcaat aatagaaaat ccatcttcat cggctttttc gtcatcatct gtatgaatca
     4141 aatcgccttc ttctgtgtca tcaaggttta attttttatg tatttctttt aacaaaccac
     4201 cataggagat taacctttta cggtgtaaac cttcctccaa atcagacaaa cgtttcaaat
     4261 tcttttcttc atcatcggtc ataaaatccg tatcctttac aggatatttt gcagtttcgt
     4321 caattgccga ttgtatatcc gatttatatt tatttttcgg tcgaatcatt tgaactttta
     4381 catttggatc atagtctaat ttcattgcct ttttccaaaa ttgaatccat tgtttttgat
     4441 tcacgtagtt ttctgtattc ttaaaataag ttggttccac acataccaat acatgcatgt
     4501 gctgattata agaattatct ttattattta ttgtcacttc cgttgcacgc ataaaaccaa
     4561 caagattttt attaattttt ttatattgca tcattcggcg aaatccttga gccatatctg
     4621 acaaactctt atttaattct tcgccatcat aaacattttt aactgttaat gtgagaaaca
     4681 accaacgaac tgttggcttt tgtttaataa cttcagcaac aaccttttgt gactgaatgc
     4741 catgtttcat tgctctcctc cagttgcaca ttggacaaag cctggattta caaaaccaca
     4801 ctcgatacaa ctttctttcg cctgtttcac gattttgttt atactctaat atttcagcac
     4861 aatcttttac tctttcagcc tttttaaatt caagaatatg cagaagttca aagtaatcaa
     4921 cattagcgat tttcttttct ctccatggtc tcacttttcc actttttgtc ttgtccacta
     4981 aaacccttga tttttcatct gaataaatgc tactattagg acacataata ttaaaagaaa
     5041 cccccatcta tttagttatt tgtttggtca cttataactt taacagatgg ggtttttctg
     5101 tgcaaccaat tttaagggtt ttcaatactt taaaacacat acataccaac acttcaacgc
     5161 acctttcagc aactaaaata aaaatgacgt tatttctata tgtatcaaga atagaaagaa
     5221 ctcgtttttc gctacgctca aaacgcaaaa aaagcactca ttcgagtgct ttttcttatc
     5281 gctccaaatc atgcgatttt ttcctctttg cttttctttg ctcacgaagt tctcgatcac
     5341 gctgcaaaac atcttgaagc gaaaaagtat tcttcttttc ttccgatcgc tcatgctgac
     5401 gcacgaaaag ccctctaggc gcataggaac aactcctaaa tgcatgtgag gggttttctc
     5461 gtccatgtga acagtcgcat acgcaatatt ttgtttccca tagctagtaa gcttggatcc
     5521 tcgcgaggcc ggccagtatt ctgacatggg tgtatcaata acccatgcgt ttccgtattg
     5581 tatcggaatg gtttcggaca gggcggtggg aatagacatg gaaaagattt ttttgcccga
     5641 ggatgcattg ataaagtatt tcttttccga aagagaggaa aagattctaa agagttttgg
     5701 aaatactgat gaatattgtg tgcagagtac aattctatgg acaagaaaag aggctttgtc
     5761 aaaacttttt cgtctgggaa tgaggatgga ttttaaaaag ctggatactt tggaggacga
     5821 ggtggttttt caggaaacaa acagggcgcg tctgttttct tttatatgca ataattactg
     5881 tatctctctg gcattgccag gttttaataa agattaaaat tattgactag aaataaaaaa
     5941 attgtccata atattaatgg acaaaaaaac aaagaattac atcaaaggaa gataaaaata
     6001 ctttgttaaa aaattaatta ttttttatct aaactattga aaatgaaaat aaaataatat
     6061 aaaatgaatc atagtgcaag agatacttgc cagaggatga atattttact gcattcatgc
     6121 tttatggcag ctaatagagg cattaaatta aattttaatt tacaatagga ggcgatatta
     6181 atgaacttta ataaaattga tttagacaat tggaagagaa aagagatatt taatcattat
     6241 ttgaaccaac aaacgacttt tagtataacc acagaaattg atattagtgt tttataccga
     6301 aacataaaac aagaaggata taaattttac cctgcattta ttttcttagt gacaagggtg
     6361 ataaactcaa atacagcttt tagaactggt tacaatagcg acggagagtt aggttattgg
     6421 gataagttag agccacttta tacaattttt gatggtgtat ctaaaacatt ctctggtatt
     6481 tggactcctg taaagaatga cttcaaagag ttttatgatt tatacctttc tgatgtagag
     6541 aaatataatg gttcggggaa attgtttccc aaaacaccta tacctgaaaa tgctttttct
     6601 ctttctatta ttccatggac ttcatttact gggtttaact taaatatcaa taataatagt
     6661 aattaccttc tacccattat tacagcagga aaattcatta ataaaggtaa ttcaatatat
     6721 ttaccgctat ctttacaggt acatcattct gtttgtgatg gttatcatgc aggattgttt
     6781 atgaactcta ttcaggaatt gtcagatagg cctaatgact ggcttttata ataaaggagg
     6841 tcgacgtcat gtttattgat acattaattg aaaagattag agaaaaggat aatccttccg
     6901 ttgtaggatt agaccctaaa attgaatatg ttccgtcttt tataaaggaa gacatgtata
     6961 aaaaatacgg gaaaaattta aaagctgtgg cagaggcgat tctcctcttc aataaatata
     7021 ttattgatgc ggtttacgat attgttcctg cagtaaaacc gcagcttgca tattatgaaa
     7081 tgtacggcct tgaaggcatg agggtgtttt atgaaacttg caaatatgca aaggaaaaag
     7141 gacttttggt tattgcagac ggaaaaagaa acgacatagg ttccaccgcc cagtgttatt
     7201 ctgccgcata tcttggaaaa acggacattg atgaaggtat aagcgaggcg gtttttgatg
     7261 tggatgccct gacagtcaac ccgtatcttg gtgtggacgg tattaagcct tttatagatg
     7321 actgtgtcaa gtacaacaag ggcatatttg ttctggtcaa gacatcaaac aagtcatccg
     7381 gagaaattca ggacatactc acccaggaag gaagaagcat ttatgagatt gttgcggagt
     7441 atgttgaatc atggggtgaa aacaaaaaag gaaaatatgg atacagttgt gtgggagcag
     7501 tggttggagc aacttatccc aatctggcca aaattttaag aaagattctg aaaaattcct
     7561 atatactggt tccgggctat ggagctcagg gaggaacagc cagagatgta gcccattgct
     7621 ttaattatga cgggctcgga gcaattgtca atgcatcaag aagcataatg tgtgcctaca
     7681 aatctgaaca atggaagaat gtttacagcg aagaaaagtt ttatgaggca tcaagagccg
     7741 aggcaataag aatgagggac gatattaaca gtgcgttgcg agacaggaag taagcccggg
     7801 cctcgagaaa acaaaaggct cagtcggaag actgggcctt ttgttttggt accgaattaa
     7861 attagaatga acgcgcgtac attattgaat aatccagtgt taaatggttt cagtttacga
     7921 tttcaaatgt ttatatccaa tttacattta aaaacataca aaacatcaaa agtatttaat
     7981 accaatattt aaaacacaat atttcaggag gaaaaactcg agttgacatc aaataagatt
     8041 cgggtcgtaa ttgctgatga taacagggaa tttggcgata ttttatatga ataccttaac
     8101 aatcaggaag atattgaagt ggtaggggtg gcaagagacg gtattgaagc ttatgagctt
     8161 atagttgaaa aacttcctga cattgctata cttgatatta tcatgcctca tttggacggt
     8221 ttgggtgttt tggagaaaat aggcgcgact gcaatcagta aaagaccttt gttcattatt
     8281 ctttcggcag tgggccagga caagataaca caaagagcat tggcactcgg agctgaatac
     8341 tatgttgtaa agcctttcga catggaagtt ttaatttcga gaataagaca actcaagaac
     8401 gtgaaccagc ccaatgttat aagacaggac ggtttgtccg gtgaagtcaa gtcgtcttat
     8461 catcctccgc agccaaaaaa tcttgaagct gaagtaacaa atattatgca tgaaataggc
     8521 gtaccggccc atataaaagg gtatcagtat ctgagggatg ccataataat ggtagtcaaa
     8581 gaccttgacg tgataaattc aattacaaag cagctgtatc ctacaatagc gaaggaatac
     8641 aacaccacac ccagcagagt tgaaagggca ataagacatg ctattgaagt ggcgtggagc
     8701 aggggacaga ttgatacaat agattcgttg tttggctata cgataaatgt tggaaaaggt
     8761 aaacccacaa attcggaatt tatagcaatg gtggcagaca aattgagatt ggaaatgaaa
     8821 atggctacat aaagatctag atctagggga ggtggacgat gaccagtgaa gaaatagcaa
     8881 aattatgtgg tgtttccaga gccacggtat ccagggttat taacaacagt cccaacgtaa
     8941 aagaggaaac gcggcaaaag attctggcag tgataaaaga aaaaaattat gttccgatag
     9001 cgccggcacg gcgtttggcc gggatagaca gcaatataat tggcctgttt gttttggata
     9061 ttgacatatc tgagtcaaag tcaagggtct cggaaagtac atacttttca cggctgataa
     9121 atctgataat agaccaggca aacaattttg gctttcaagt attggtgtca attataactt
     9181 cacagaaaca gctgagtgaa attagaaatc tcttcatgag cagaaccatt ttcagcggca
     9241 tttttatcgg tgcgttcaat gatgaaatcc aacttgatga tgatattata atgcaacatc
     9301 ccacaattat tattgaccgc caatcagaaa ggatggtgaa aaagccaaac agattggttg
     9361 taaacctgga caactttgag ggtgcttata atgcgacaca gtttttgatt aaattggggc
     9421 ataccagaat tgggcacata tccggggacc ttagaaaact ttcgggcata gaacgctatg
     9481 aaggatacaa aaaagcattg gaagatgcag gattaggttt tgacaaaaat ttggttcgtg
     9541 aagggaactt ccttgatgac agcggctata ggcttgcacg tgagatatta aaagagaacg
     9601 tgacggctat tttctgtgcc aatgatgtaa tggcaattag tgcaattaaa gccataaaag
     9661 aaacgggttt gagtgtaccg gatgatatat ctgtaatagg gtttgataat acagcaatcg
     9721 gaaattatat catgcctgca ttgacaactg tgaacgcgcc gttggagcat attgcagaag
     9781 catgtattga gtcattgaaa tacttttgcg agcacaaaca ttttaaacaa aaggaaatca
     9841 gggttaaaac cgatttgata atccgggatt caaccaagag ggctttggaa ttctgacccg
     9901 g
//
