LOCUS       V007677                 9103 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V007677
VERSION     V007677
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9103)
  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 9103)
  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 9103)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9103)
  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..9103
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             22..1059
                     /codon_start=1
                     /gene="glyR3"
                     /product="GlyR3"
                     /label="glyR3"
                     /note="repressor"
                     /protein_id="AGT95718.1"
                     /translation="MTSEEIAKLCGVSRATVSRVINNSPNVKEETRQKILAVIKEKNYV
                     PIAPARRLAGIDSNIIGLFVLDIDISESKSRVSESTYFSRLINLIIDQANNFGFQVLVS
                     IITSQKQLSEIRNLFMSRTIFSGIFIGAFNDEIQLDDDIIMQHPTIIIDRQSERMVKKP
                     NRLVVNLDNFEGAYNATQFLIKLGHTRIGHISGDLRKLSGIERYEGYKKALEDAGLGFD
                     KNLVREGNFLDDSGYRLAREILKENVTAIFCANDVMAISAIKAIKETGLSVPDDISVIG
                     FDNTAIGNYIMPALTTVNAPLEHIAEACIESLKYFCEHKHFKQKEIRVKTDLIIRDSTK
                     RALEF"
     gene            22..1059
                     /gene="glyR3"
                     /label="glyR3"
     terminator      1114..1361
                     /label="CYC1 terminator"
                     /note="transcription terminator for CYC1"
     rep_origin      complement(1609..2197)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(2371..3228)
                     /label="AmpR"
                     /note="beta-lactamase"
     CDS             complement(3327..4127)
                     /label="URA3"
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     promoter        complement(4128..4348)
                     /label="URA3 promoter"
     misc_feature    4376..4879
                     /label="CEN/ARS"
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     CDS             complement(5111..6112)
                     /label="repB"
                     /note="RepB replication protein"
     CDS             7246..7893
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     regulatory      8938..8976
                     /gene="celC"
                     /regulatory_class="promoter"
     gene            8938..8976
                     /gene="celC"
                     /label="celC"
ORIGIN
        1 gagatctagg ggaggtggac gatgaccagt gaagaaatag caaaattatg tggtgtttcc
       61 agagccacgg tatccagggt tattaacaac agtcccaacg taaaagagga aacgcggcaa
      121 aagattctgg cagtgataaa agaaaaaaat tatgttccga tagcgccggc acggcgtttg
      181 gccgggatag acagcaatat aattggcctg tttgttttgg atattgacat atctgagtca
      241 aagtcaaggg tctcggaaag tacatacttt tcacggctga taaatctgat aatagaccag
      301 gcaaacaatt ttggctttca agtattggtg tcaattataa cttcacagaa acagctgagt
      361 gaaattagaa atctcttcat gagcagaacc attttcagcg gcatttttat cggtgcgttc
      421 aatgatgaaa tccaacttga tgatgatatt ataatgcaac atcccacaat tattattgac
      481 cgccaatcag aaaggatggt gaaaaagcca aacagattgg ttgtaaacct ggacaacttt
      541 gagggtgctt ataatgcgac acagtttttg attaaattgg ggcataccag aattgggcac
      601 atatccgggg accttagaaa actttcgggc atagaacgct atgaaggata caaaaaagca
      661 ttggaagatg caggattagg ttttgacaaa aatttggttc gtgaagggaa cttccttgat
      721 gacagcggct ataggcttgc acgtgagata ttaaaagaga acgtgacggc tattttctgt
      781 gccaatgatg taatggcaat tagtgcaatt aaagccataa aagaaacggg tttgagtgta
      841 ccggatgata tatctgtaat agggtttgat aatacagcaa tcggaaatta tatcatgcct
      901 gcattgacaa ctgtgaacgc gccgttggag catattgcag aagcatgtat tgagtcattg
      961 aaatactttt gcgagcacaa acattttaaa caaaaggaaa tcagggttaa aaccgatttg
     1021 ataatccggg attcaaccaa gagggctttg gaattctgac ccgggaattc ggcgcgcctc
     1081 agcgtttaaa cccgctgatc ctagagggcc gcatcatgta attagttatg tcacgcttac
     1141 attcacgccc tccccccaca tccgctctaa ccgaaaagga aggagttaga caacctgaag
     1201 tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta tttatatttc
     1261 aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata ctgaaaacct
     1321 tgcttgagaa ggttttggga cgctcgaagg ctttaatttg caagctgcgg ccctgcatta
     1381 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     1441 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     1501 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     1561 aggccagcaa aagcccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1621 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1681 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1741 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1801 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1861 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1921 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     1981 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     2041 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     2101 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     2161 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     2221 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     2281 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     2341 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     2401 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     2461 gatacgggag cgcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     2521 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     2581 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     2641 tagttcgcca gttaatagtt tgcgcaacgt tgttggcatt gctacaggca tcgtggtgtc
     2701 actctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     2761 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     2821 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     2881 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     2941 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataatagtgt
     3001 atcacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     3061 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     3121 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     3181 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     3241 tcaatgggta ataactgata taattaaatt gaagctctaa tttgtgagtt tagtatacat
     3301 gcatttactt ataatacagt tttttagttt tgctggccgc atcttctcaa atatgcttcc
     3361 cagcctgctt ttctgtaacg ttcaccctct accttagcat cccttccctt tgcaaatagt
     3421 cctcttccaa caataataat gtcagatcct gtagagacca catcatccac ggttctatac
     3481 tgttgaccca atgcgtctcc cttgtcatct aaacccacac cgggtgtcat aatcaaccaa
     3541 tcgtaacctt catctcttcc acccatgtct ctttgagcaa taaagccgat aacaaaatct
     3601 ttgtcgctct tcgcaatgtc aacagtaccc ttagtatatt ctccagtaga tagggagccc
     3661 ttgcatgaca attctgctaa catcaaaagg cctctaggtt cctttgttac ttcttctgcc
     3721 gcctgcttca aaccgctaac aatacctggg cccaccacac cgtgtgcatt cgtaatgtct
     3781 gcccattctg ctattctgta tacacccgca gagtactgca atttgactgt attaccaatg
     3841 tcagcaaatt ttctgtcttc gaagagtaaa aaattgtact tggcggataa tgcctttagc
     3901 ggcttaactg tgccctccat ggaaaaatca gtcaagatat ccacatgtgt ttttagtaaa
     3961 caaattttgg gacctaatgc ttcaactaac tccagtaatt ccttggtggt acgaacatcc
     4021 aatgaagcac acaagtttgt ttgcttttcg tgcatgatat taaatagctt ggcagcaaca
     4081 ggactaggat gagtagcagc acgttcctta tatgtagctt tcgacatgat ttatcttcgt
     4141 ttcctgcagg tttttgttct gtgcagttgg gttaagaata ctgggcaatt tcatgtttct
     4201 tcaacactac atatgcgtat atataccaat ctaagtctgt gctccttcct tcgttcttcc
     4261 ttctgttcgg agattaccga atcaaaaaaa tttcaaagaa accgaaatca aaaaaaagaa
     4321 taaaaaaaaa atgatgaatt gaattgaaaa gctagcttat cgatgggtcc ttttcatcac
     4381 gtgctataaa aataattata atttaaattt tttaatataa atatataaat taaaaataga
     4441 aagtaaaaaa agaaattaaa gaaaaaatag tttttgtttt ccgaagatgt aaaagactct
     4501 agggggatcg ccaacaaata ctacctttta tcttgctctt cctgctctca ggtattaatg
     4561 ccgaattgtt tcatcttgtc tgtgtagaag accacacacg aaaatcctgt gattttacat
     4621 tttacttatc gttaatcgaa tgtatatcta tttaatctgc ttttcttgtc taataaatat
     4681 atatgtaaag tacgcttttt gttgaaattt tttaaacctt tgtttatttt tttttcttca
     4741 ttccgtaact cttctacctt ctttatttac tttctaaaat ccaaatacaa aacataaaaa
     4801 taaataaaca cagagtaaat tcccaaatta ttccatcatt aaaagatacg aggcgcgtgt
     4861 aagttacagg caagcgatcc gtccgccggc gaacgtggcg agaaaggaag ggaagaaagc
     4921 gaaaggagcg ggggctaggg cggtgggaag tgtaggggtc acgctgggcg taaccaccac
     4981 acccgccgcg cttaatgggg cgctacaggg cgcgtgggga tgatccacta gtgaatttag
     5041 gaggcttact tgtctgcttt cttcattaga atcaatcctt ttttaaaagt caatcccgtt
     5101 tgttgaacta ctctttaata aaataatttt tccgttccca attccacatt gcaataatag
     5161 aaaatccatc ttcatcggct ttttcgtcat catctgtatg aatcaaatcg ccttcttctg
     5221 tgtcatcaag gtttaatttt ttatgtattt cttttaacaa accaccatag gagattaacc
     5281 ttttacggtg taaaccttcc tccaaatcag acaaacgttt caaattcttt tcttcatcat
     5341 cggtcataaa atccgtatcc tttacaggat attttgcagt ttcgtcaatt gccgattgta
     5401 tatccgattt atatttattt ttcggtcgaa tcatttgaac ttttacattt ggatcatagt
     5461 ctaatttcat tgcctttttc caaaattgaa tccattgttt ttgattcacg tagttttctg
     5521 tattcttaaa ataagttggt tccacacata ccaatacatg catgtgctga ttataagaat
     5581 tatctttatt atttattgtc acttccgttg cacgcataaa accaacaaga tttttattaa
     5641 tttttttata ttgcatcatt cggcgaaatc cttgagccat atctgacaaa ctcttattta
     5701 attcttcgcc atcataaaca tttttaactg ttaatgtgag aaacaaccaa cgaactgttg
     5761 gcttttgttt aataacttca gcaacaacct tttgtgactg aatgccatgt ttcattgctc
     5821 tcctccagtt gcacattgga caaagcctgg atttacaaaa ccacactcga tacaactttc
     5881 tttcgcctgt ttcacgattt tgtttatact ctaatatttc agcacaatct tttactcttt
     5941 cagccttttt aaattcaaga atatgcagaa gttcaaagta atcaacatta gcgattttct
     6001 tttctctcca tggtctcact tttccacttt ttgtcttgtc cactaaaacc cttgattttt
     6061 catctgaata aatgctacta ttaggacaca taatattaaa agaaaccccc atctatttag
     6121 ttatttgttt ggtcacttat aactttaaca gatggggttt ttctgtgcaa ccaattttaa
     6181 gggttttcaa tactttaaaa cacatacata ccaacacttc aacgcacctt tcagcaacta
     6241 aaataaaaat gacgttattt ctatatgtat caagaataga aagaactcgt ttttcgctac
     6301 gctcaaaacg caaaaaaagc actcattcga gtgctttttc ttatcgctcc aaatcatgcg
     6361 attttttcct ctttgctttt ctttgctcac gaagttctcg atcacgctgc aaaacatctt
     6421 gaagcgaaaa agtattcttc ttttcttccg atcgctcatg ctgacgcacg aaaagccctc
     6481 taggcgcata ggaacaactc ctaaatgcat gtgaggggtt ttctcgtcca tgtgaacagt
     6541 cgcatacgca atattttgtt tcccatagct agtaagcttg gatcctcgcg aggccggcca
     6601 gtattctgac atgggtgtat caataaccca tgcgtttccg tattgtatcg gaatggtttc
     6661 ggacagggcg gtgggaatag acatggaaaa gatttttttg cccgaggatg cattgataaa
     6721 gtatttcttt tccgaaagag aggaaaagat tctaaagagt tttggaaata ctgatgaata
     6781 ttgtgtgcag agtacaattc tatggacaag aaaagaggct ttgtcaaaac tttttcgtct
     6841 gggaatgagg atggatttta aaaagctgga tactttggag gacgaggtgg tttttcagga
     6901 aacaaacagg gcgcgtctgt tttcttttat atgcaataat tactgtatct ctctggcatt
     6961 gccaggtttt aataaagatt aaaattattg actagaaata aaaaaattgt ccataatatt
     7021 aatggacaaa aaaacaaaga attacatcaa aggaagataa aaatactttg ttaaaaaatt
     7081 aattattttt tatctaaact attgaaaatg aaaataaaat aatataaaat gaatcatagt
     7141 gcaagagata cttgccagag gatgaatatt ttactgcatt catgctttat ggcagctaat
     7201 agaggcatta aattaaattt taatttacaa taggaggcga tattaatgaa ctttaataaa
     7261 attgatttag acaattggaa gagaaaagag atatttaatc attatttgaa ccaacaaacg
     7321 acttttagta taaccacaga aattgatatt agtgttttat accgaaacat aaaacaagaa
     7381 ggatataaat tttaccctgc atttattttc ttagtgacaa gggtgataaa ctcaaataca
     7441 gcttttagaa ctggttacaa tagcgacgga gagttaggtt attgggataa gttagagcca
     7501 ctttatacaa tttttgatgg tgtatctaaa acattctctg gtatttggac tcctgtaaag
     7561 aatgacttca aagagtttta tgatttatac ctttctgatg tagagaaata taatggttcg
     7621 gggaaattgt ttcccaaaac acctatacct gaaaatgctt tttctctttc tattattcca
     7681 tggacttcat ttactgggtt taacttaaat atcaataata atagtaatta ccttctaccc
     7741 attattacag caggaaaatt cattaataaa ggtaattcaa tatatttacc gctatcttta
     7801 caggtacatc attctgtttg tgatggttat catgcaggat tgtttatgaa ctctattcag
     7861 gaattgtcag ataggcctaa tgactggctt ttataataaa ggaggtcgac gtcatgttta
     7921 ttgatacatt aattgaaaag attagagaaa aggataatcc ttccgttgta ggattagacc
     7981 ctaaaattga atatgttccg tcttttataa aggaagacat gtataaaaaa tacgggaaaa
     8041 atttaaaagc tgtggcagag gcgattctcc tcttcaataa atatattatt gatgcggttt
     8101 acgatattgt tcctgcagta aaaccgcagc ttgcatatta tgaaatgtac ggccttgaag
     8161 gcatgagggt gttttatgaa acttgcaaat atgcaaagga aaaaggactt ttggttattg
     8221 cagacggaaa aagaaacgac ataggttcca ccgcccagtg ttattctgcc gcatatcttg
     8281 gaaaaacgga cattgatgaa ggtataagcg aggcggtttt tgatgtggat gccctgacag
     8341 tcaacccgta tcttggtgtg gacggtatta agccttttat agatgactgt gtcaagtaca
     8401 acaagggcat atttgttctg gtcaagacat caaacaagtc atccggagaa attcaggaca
     8461 tactcaccca ggaaggaaga agcatttatg agattgttgc ggagtatgtt gaatcatggg
     8521 gtgaaaacaa aaaaggaaaa tatggataca gttgtgtggg agcagtggtt ggagcaactt
     8581 atcccaatct ggccaaaatt ttaagaaaga ttctgaaaaa ttcctatata ctggttccgg
     8641 gctatggagc tcagggagga acagccagag atgtagccca ttgctttaat tatgacgggc
     8701 tcggagcaat tgtcaatgca tcaagaagca taatgtgtgc ctacaaatct gaacaatgga
     8761 agaatgttta cagcgaagaa aagttttatg aggcatcaag agccgaggca ataagaatga
     8821 gggacgatat taacagtgcg ttgcgagaca ggaagtaagc ccgggcctcg agaaaacaaa
     8881 aggctcagtc ggaagactgg gccttttgtt ttggtaccgg aattcgagct cgcatgcaat
     8941 taaattagaa tgaacgcgcg tacattattg aataatccag tgttaaatgg tttcagttta
     9001 cgatttcaaa tgtttatatc caatttacat ttaaaaacat acaaaacatc aaaagtattt
     9061 aataccaata tttaaaacac aatatttcag gaggaaaaac tcg
//
