LOCUS       V007676                 9485 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V007676
VERSION     V007676
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9485)
  AUTHORS   Mearls EB, Olson DG, Herring CD, Lynd LR.
  TITLE     Development of a regulatable plasmid-based gene expression system
            for Clostridium thermocellum
  JOURNAL   Appl. Microbiol. Biotechnol. 99 (18), 7589-7599 (2015)
   PUBMED   25994254
REFERENCE   2  (bases 1 to 9485)
  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 9485)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9485)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
            Microbiol. Biotechnol."; date: "2015"; volume: "99"; issue: "18";
            pages: "7589-7599"
            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..9485
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      57..304
                     /label="CYC1 terminator"
                     /note="transcription terminator for CYC1"
     rep_origin      complement(552..1140)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(1314..2171)
                     /label="AmpR"
                     /note="beta-lactamase"
     CDS             complement(2270..3070)
                     /label="URA3"
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     promoter        complement(3071..3291)
                     /label="URA3 promoter"
     misc_feature    3319..3822
                     /label="CEN/ARS"
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     CDS             complement(4054..5055)
                     /label="repB"
                     /note="RepB replication protein"
     regulatory      5541..6188
                     /gene="gapD"
                     /regulatory_class="promoter"
     gene            5541..6188
                     /gene="gapD"
                     /label="gapD"
     CDS             6189..6836
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     regulatory      7417..7456
                     /regulatory_class="terminator"
     regulatory      7464..7502
                     /gene="celC"
                     /regulatory_class="promoter"
     gene            7464..7502
                     /gene="celC"
                     /label="celC"
     CDS             7626..8663
                     /codon_start=1
                     /gene="glyR3"
                     /product="GlyR3"
                     /label="glyR3"
                     /note="repressor"
                     /protein_id="AGT95728.1"
                     /translation="MTSEEIAKLCGVSRATVSRVINNSPNVKEETRQKILAVIKEKNYV
                     PIAPARRLAGIDSNIIGLFVLDIDISESKSRVSESTYFSRLINLIIDQANNFGFQVLVS
                     IITSQKQLSEIRNLFMSRTIFSGIFIGAFNDEIQLDDDIIMQHPTIIIDRQSERMVKKP
                     NRLVVNLDNFEGAYNATQFLIKLGHTRIGHISGDLRKLSGIERYEGYKKALEDAGLGFD
                     KNLVREGNFLDDSGYRLAREILKENVTAIFCANDVMAISAIKAIKETGLSVPDDISVIG
                     FDNTAIGNYIMPALTTVNAPLEHIAEACIESLKYFCEHKHFKQKEIRVKTDLIIRDSTK
                     RALEF"
     gene            7626..8663
                     /gene="glyR3"
                     /label="glyR3"
     CDS             8676..9485
                     /codon_start=1
                     /gene="spo0A"
                     /product="Spo0A"
                     /label="spo0A"
                     /note="sporulation response regulator A"
                     /protein_id="AGT95729.1"
                     /translation="MTSNKIRVVIADDNREFGDILYEYLNNQEDIEVVGVARDGIEAYE
                     LIVEKLPDIAILDIIMPHLDGLGVLEKIGATAISKRPLFIILSAVGQDKITQRALALGA
                     EYYVVKPFDMEVLISRIRQLKNVNQPNVIRQDGLSGEVKSSYHPPQPKNLEAEVTNIMH
                     EIGVPAHIKGYQYLRDAIIMVVKDLDVINSITKQLYPTIAKEYNTTPSRVERAIRHAIE
                     VAWSRGQIDTIDSLFGYTINVGKGKPTNSEFIAMVADKLRLEMKMAT"
     gene            8676..9485
                     /gene="spo0A"
                     /label="spo0A"
ORIGIN
        1 gcggccgcaa ttcggcgcgc ctcagcgttt aaacccgctg atcctagagg gccgcatcat
       61 gtaattagtt atgtcacgct tacattcacg ccctcccccc acatccgctc taaccgaaaa
      121 ggaaggagtt agacaacctg aagtctaggt ccctatttat ttttttatag ttatgttagt
      181 attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc
      241 atgtaacatt atactgaaaa ccttgcttga gaaggttttg ggacgctcga aggctttaat
      301 ttgcaagctg cggccctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
      361 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
      421 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
      481 gcaggaaaga acatgtgagc aaaaggccag caaaagccca ggaaccgtaa aaaggccgcg
      541 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
      601 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
      661 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
      721 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
      781 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
      841 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
      901 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
      961 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     1021 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     1081 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     1141 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     1201 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     1261 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     1321 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     1381 ctccccgtcg tgtagataac tacgatacgg gagcgcttac catctggccc cagtgctgca
     1441 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     1501 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     1561 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttggc
     1621 attgctacag gcatcgtggt gtcactctcg tcgtttggta tggcttcatt cagctccggt
     1681 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     1741 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     1801 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     1861 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     1921 gcgtcaatac gggataatag tgtatcacat agcagaactt taaaagtgct catcattgga
     1981 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     2041 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     2101 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     2161 tgaatactca tactcttcct ttttcaatgg gtaataactg atataattaa attgaagctc
     2221 taatttgtga gtttagtata catgcattta cttataatac agttttttag ttttgctggc
     2281 cgcatcttct caaatatgct tcccagcctg cttttctgta acgttcaccc tctaccttag
     2341 catcccttcc ctttgcaaat agtcctcttc caacaataat aatgtcagat cctgtagaga
     2401 ccacatcatc cacggttcta tactgttgac ccaatgcgtc tcccttgtca tctaaaccca
     2461 caccgggtgt cataatcaac caatcgtaac cttcatctct tccacccatg tctctttgag
     2521 caataaagcc gataacaaaa tctttgtcgc tcttcgcaat gtcaacagta cccttagtat
     2581 attctccagt agatagggag cccttgcatg acaattctgc taacatcaaa aggcctctag
     2641 gttcctttgt tacttcttct gccgcctgct tcaaaccgct aacaatacct gggcccacca
     2701 caccgtgtgc attcgtaatg tctgcccatt ctgctattct gtatacaccc gcagagtact
     2761 gcaatttgac tgtattacca atgtcagcaa attttctgtc ttcgaagagt aaaaaattgt
     2821 acttggcgga taatgccttt agcggcttaa ctgtgccctc catggaaaaa tcagtcaaga
     2881 tatccacatg tgtttttagt aaacaaattt tgggacctaa tgcttcaact aactccagta
     2941 attccttggt ggtacgaaca tccaatgaag cacacaagtt tgtttgcttt tcgtgcatga
     3001 tattaaatag cttggcagca acaggactag gatgagtagc agcacgttcc ttatatgtag
     3061 ctttcgacat gatttatctt cgtttcctgc aggtttttgt tctgtgcagt tgggttaaga
     3121 atactgggca atttcatgtt tcttcaacac tacatatgcg tatatatacc aatctaagtc
     3181 tgtgctcctt ccttcgttct tccttctgtt cggagattac cgaatcaaaa aaatttcaaa
     3241 gaaaccgaaa tcaaaaaaaa gaataaaaaa aaaatgatga attgaattga aaagctagct
     3301 tatcgatggg tccttttcat cacgtgctat aaaaataatt ataatttaaa ttttttaata
     3361 taaatatata aattaaaaat agaaagtaaa aaaagaaatt aaagaaaaaa tagtttttgt
     3421 tttccgaaga tgtaaaagac tctaggggga tcgccaacaa atactacctt ttatcttgct
     3481 cttcctgctc tcaggtatta atgccgaatt gtttcatctt gtctgtgtag aagaccacac
     3541 acgaaaatcc tgtgatttta cattttactt atcgttaatc gaatgtatat ctatttaatc
     3601 tgcttttctt gtctaataaa tatatatgta aagtacgctt tttgttgaaa ttttttaaac
     3661 ctttgtttat ttttttttct tcattccgta actcttctac cttctttatt tactttctaa
     3721 aatccaaata caaaacataa aaataaataa acacagagta aattcccaaa ttattccatc
     3781 attaaaagat acgaggcgcg tgtaagttac aggcaagcga tccgtccgcc ggcgaacgtg
     3841 gcgagaaagg aagggaagaa agcgaaagga gcgggggcta gggcggtggg aagtgtaggg
     3901 gtcacgctgg gcgtaaccac cacacccgcc gcgcttaatg gggcgctaca gggcgcgtgg
     3961 ggatgatcca ctagtgaatt taggaggctt acttgtctgc tttcttcatt agaatcaatc
     4021 cttttttaaa agtcaatccc gtttgttgaa ctactcttta ataaaataat ttttccgttc
     4081 ccaattccac attgcaataa tagaaaatcc atcttcatcg gctttttcgt catcatctgt
     4141 atgaatcaaa tcgccttctt ctgtgtcatc aaggtttaat tttttatgta tttcttttaa
     4201 caaaccacca taggagatta accttttacg gtgtaaacct tcctccaaat cagacaaacg
     4261 tttcaaattc ttttcttcat catcggtcat aaaatccgta tcctttacag gatattttgc
     4321 agtttcgtca attgccgatt gtatatccga tttatattta tttttcggtc gaatcatttg
     4381 aacttttaca tttggatcat agtctaattt cattgccttt ttccaaaatt gaatccattg
     4441 tttttgattc acgtagtttt ctgtattctt aaaataagtt ggttccacac ataccaatac
     4501 atgcatgtgc tgattataag aattatcttt attatttatt gtcacttccg ttgcacgcat
     4561 aaaaccaaca agatttttat taattttttt atattgcatc attcggcgaa atccttgagc
     4621 catatctgac aaactcttat ttaattcttc gccatcataa acatttttaa ctgttaatgt
     4681 gagaaacaac caacgaactg ttggcttttg tttaataact tcagcaacaa ccttttgtga
     4741 ctgaatgcca tgtttcattg ctctcctcca gttgcacatt ggacaaagcc tggatttaca
     4801 aaaccacact cgatacaact ttctttcgcc tgtttcacga ttttgtttat actctaatat
     4861 ttcagcacaa tcttttactc tttcagcctt tttaaattca agaatatgca gaagttcaaa
     4921 gtaatcaaca ttagcgattt tcttttctct ccatggtctc acttttccac tttttgtctt
     4981 gtccactaaa acccttgatt tttcatctga ataaatgcta ctattaggac acataatatt
     5041 aaaagaaacc cccatctatt tagttatttg tttggtcact tataacttta acagatgggg
     5101 tttttctgtg caaccaattt taagggtttt caatacttta aaacacatac ataccaacac
     5161 ttcaacgcac ctttcagcaa ctaaaataaa aatgacgtta tttctatatg tatcaagaat
     5221 agaaagaact cgtttttcgc tacgctcaaa acgcaaaaaa agcactcatt cgagtgcttt
     5281 ttcttatcgc tccaaatcat gcgatttttt cctctttgct tttctttgct cacgaagttc
     5341 tcgatcacgc tgcaaaacat cttgaagcga aaaagtattc ttcttttctt ccgatcgctc
     5401 atgctgacgc acgaaaagcc ctctaggcgc ataggaacaa ctcctaaatg catgtgaggg
     5461 gttttctcgt ccatgtgaac agtcgcatac gcaatatttt gtttcccata gctagtaagc
     5521 ttggatcctc gcgaggccgg ccagtattct gacatgggtg tatcaataac ccatgcgttt
     5581 ccgtattgta tcggaatggt ttcggacagg gcggtgggaa tagacatgga aaagattttt
     5641 ttgcccgagg atgcattgat aaagtatttc ttttccgaaa gagaggaaaa gattctaaag
     5701 agttttggaa atactgatga atattgtgtg cagagtacaa ttctatggac aagaaaagag
     5761 gctttgtcaa aactttttcg tctgggaatg aggatggatt ttaaaaagct ggatactttg
     5821 gaggacgagg tggtttttca ggaaacaaac agggcgcgtc tgttttcttt tatatgcaat
     5881 aattactgta tctctctggc attgccaggt tttaataaag attaaaatta ttgactagaa
     5941 ataaaaaaat tgtccataat attaatggac aaaaaaacaa agaattacat caaaggaaga
     6001 taaaaatact ttgttaaaaa attaattatt ttttatctaa actattgaaa atgaaaataa
     6061 aataatataa aatgaatcat agtgcaagag atacttgcca gaggatgaat attttactgc
     6121 attcatgctt tatggcagct aatagaggca ttaaattaaa ttttaattta caataggagg
     6181 cgatattaat gaactttaat aaaattgatt tagacaattg gaagagaaaa gagatattta
     6241 atcattattt gaaccaacaa acgactttta gtataaccac agaaattgat attagtgttt
     6301 tataccgaaa cataaaacaa gaaggatata aattttaccc tgcatttatt ttcttagtga
     6361 caagggtgat aaactcaaat acagctttta gaactggtta caatagcgac ggagagttag
     6421 gttattggga taagttagag ccactttata caatttttga tggtgtatct aaaacattct
     6481 ctggtatttg gactcctgta aagaatgact tcaaagagtt ttatgattta tacctttctg
     6541 atgtagagaa atataatggt tcggggaaat tgtttcccaa aacacctata cctgaaaatg
     6601 ctttttctct ttctattatt ccatggactt catttactgg gtttaactta aatatcaata
     6661 ataatagtaa ttaccttcta cccattatta cagcaggaaa attcattaat aaaggtaatt
     6721 caatatattt accgctatct ttacaggtac atcattctgt ttgtgatggt tatcatgcag
     6781 gattgtttat gaactctatt caggaattgt cagataggcc taatgactgg cttttataat
     6841 aaaggaggtc gacgtcatgg aaaatttatc aaaagacatc gatgaaattt tgatcacaga
     6901 agaagaactt aaggaaaaga taaaagagct tgggaggcaa atcacaaaag actacaaagg
     6961 gaaaaatttg atgttggtag gagttttaaa aggtgcttta atgtttatgg ctgatttgtc
     7021 aagacacata gatttgcctt tatcacttga ttttatggct gtttccagct atggaagctc
     7081 aactcattca tcaggaatag taaagataat caaagatctt gatataagca tagaaggcaa
     7141 agatgttctg attgtggaag acataattga cagcggtttg actttgtctt acttaaggga
     7201 aactttactt ggaaggaagc caaaaagcct gaaaatatgc acaatattag acaaaccgga
     7261 gagaagagaa gcatctgtaa aagtcgatta tgtaggattt aagatacctg ataagtttgt
     7321 cgtgggttat ggattggact ttgatgaaaa gtacaggaac cttcctttta taggcgtttt
     7381 gaaacctgaa atgtacagct aagcccgggc ctcgagaaaa caaaaggctc agtcggaaga
     7441 ctgggccttt tgttttggta ccgaattaaa ttagaatgaa cgcgcgtaca ttattgaata
     7501 atccagtgtt aaatggtttc agtttacgat ttcaaatgtt tatatccaat ttacatttaa
     7561 aaacatacaa aacatcaaaa gtatttaata ccaatattta aaacacaata tttcaggagg
     7621 aaaaaatgac cagtgaagaa atagcaaaat tatgtggtgt ttccagagcc acggtatcca
     7681 gggttattaa caacagtccc aacgtaaaag aggaaacgcg gcaaaagatt ctggcagtga
     7741 taaaagaaaa aaattatgtt ccgatagcgc cggcacggcg tttggccggg atagacagca
     7801 atataattgg cctgtttgtt ttggatattg acatatctga gtcaaagtca agggtctcgg
     7861 aaagtacata cttttcacgg ctgataaatc tgataataga ccaggcaaac aattttggct
     7921 ttcaagtatt ggtgtcaatt ataacttcac agaaacagct gagtgaaatt agaaatctct
     7981 tcatgagcag aaccattttc agcggcattt ttatcggtgc gttcaatgat gaaatccaac
     8041 ttgatgatga tattataatg caacatccca caattattat tgaccgccaa tcagaaagga
     8101 tggtgaaaaa gccaaacaga ttggttgtaa acctggacaa ctttgagggt gcttataatg
     8161 cgacacagtt tttgattaaa ttggggcata ccagaattgg gcacatatcc ggggacctta
     8221 gaaaactttc gggcatagaa cgctatgaag gatacaaaaa agcattggaa gatgcaggat
     8281 taggttttga caaaaatttg gttcgtgaag ggaacttcct tgatgacagc ggctataggc
     8341 ttgcacgtga gatattaaaa gagaacgtga cggctatttt ctgtgccaat gatgtaatgg
     8401 caattagtgc aattaaagcc ataaaagaaa cgggtttgag tgtaccggat gatatatctg
     8461 taatagggtt tgataataca gcaatcggaa attatatcat gcctgcattg acaactgtga
     8521 acgcgccgtt ggagcatatt gcagaagcat gtattgagtc attgaaatac ttttgcgagc
     8581 acaaacattt taaacaaaag gaaatcaggg ttaaaaccga tttgataatc cgggattcaa
     8641 ccaagagggc tttggaattc tgaaggaggg catgcttgac atcaaataag attcgggtcg
     8701 taattgctga tgataacagg gaatttggcg atattttata tgaatacctt aacaatcagg
     8761 aagatattga agtggtaggg gtggcaagag acggtattga agcttatgag cttatagttg
     8821 aaaaacttcc tgacattgct atacttgata ttatcatgcc tcatttggac ggtttgggtg
     8881 ttttggagaa aataggcgcg actgcaatca gtaaaagacc tttgttcatt attctttcgg
     8941 cagtgggcca ggacaagata acacaaagag cattggcact cggagctgaa tactatgttg
     9001 taaagccttt cgacatggaa gttttaattt cgagaataag acaactcaag aacgtgaacc
     9061 agcccaatgt tataagacag gacggtttgt ccggtgaagt caagtcgtct tatcatcctc
     9121 cgcagccaaa aaatcttgaa gctgaagtaa caaatattat gcatgaaata ggcgtaccgg
     9181 cccatataaa agggtatcag tatctgaggg atgccataat aatggtagtc aaagaccttg
     9241 acgtgataaa ttcaattaca aagcagctgt atcctacaat agcgaaggaa tacaacacca
     9301 cacccagcag agttgaaagg gcaataagac atgctattga agtggcgtgg agcaggggac
     9361 agattgatac aatagattcg ttgtttggct atacgataaa tgttggaaaa ggtaaaccca
     9421 caaattcgga atttatagca atggtggcag acaaattgag attggaaatg aaaatggcta
     9481 cataa
//
