LOCUS       V004404                 5650 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004404
VERSION     V004404
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5650)
  AUTHORS   Olson DG, Lynd LR.
  TITLE     Computational Design and Characterization of a Temperature-sensitive
            Replicon for Gram Positive Thermophiles
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5650)
  AUTHORS   Olson DG, Lynd LR.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-FEB-2011) Thayer School of Engineering, Dartmouth
            College, 8000 Cummings, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 5650)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5650)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (23-FEB-2011) Thayer School of Engineering, Dartmouth College, 8000
            Cummings, Hanover, NH 03755, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5650
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             1281..2282
                     /label="repB"
                     /note="RepB replication protein"
     promoter        2400..2504
                     /label="AmpR promoter"
     CDS             2505..3362
                     /label="AmpR"
                     /note="beta-lactamase"
     protein_bind    3524..3540
                     /label="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     3548..3564
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     terminator      3656..3683
                     /label="T7Te terminator"
                     /note="phage T7 early transcription terminator"
     CDS             complement(3767..4414)
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     regulatory      complement(4415..4942)
                     /gene="gapDH"
                     /label="derived from Clostridium thermocellum DSM1313"
                     /note="derived from Clostridium thermocellum DSM1313"
                     /regulatory_class="promoter"
     gene            complement(4415..4942)
                     /gene="gapDH"
                     /label="gapDH"
                     /note="derived from Clostridium thermocellum DSM1313"
     terminator      complement(5049..5076)
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(5168..5249)
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagtatgg gaaacaaaat
      841 attgcgtatg cgactgttca catggacgag aaaacccctc acatgcattt aggagttgtt
      901 cctatgcgcc tagagggctt ttcgtgcgtc agcatgagcg atcggaagaa aagaagaata
      961 ctttttcgct tcaagatgtt ttgcagcgtg atcgagaact tcgtgagcaa agaaaagcaa
     1021 agaggaaaaa atcgcatgat ttggagcgat aagaaaaagc actcgaatga gtgctttttt
     1081 tgcgttttga gcgtagcgaa aaacgagttc tttctattct tgatacatat agaaataacg
     1141 tcatttttat tttagttgct gaaaggtgcg ttgacgcgtt ggtatgtatg tgttttaaag
     1201 tattgaaaac ccttaaaatt ggttgcacag aaaaacccca tctgttaaag ttataagtga
     1261 ccaaacaaat aactaaatag atgggggttt cttttaatat tatgtgtcct aatagtagca
     1321 tttattcaga tgaaaaatca agggttttag tggacaagac aaaaagtgga aaagtgagac
     1381 catggagaga aaagaaaatc gctaatgttg attactttga acttctgcat attcttgaat
     1441 ttaaaaaggc tgaaagagta aaagattgtg ctgaaatatt agagtataaa caaaatcgtg
     1501 aaacaggcga aagaaagttg tatcgagtgt ggttttgtaa atccaggctt tgtccaatgt
     1561 gcaactggag gagagcaatg aaacatggca ttcagtcaca aaaggttgtt gctgaagtta
     1621 ttaaacaaaa gccaacagtt cgttggttgt ttctcacatt aacagttaaa aatgtttatg
     1681 atggcgaaga attaaataag agtttgtcag atatggctca aggatttcgc cgaatgatgc
     1741 aatataaaaa aattaataaa aatcttgttg gttttatgcg tgcaacggaa gtgacaataa
     1801 ataataaaga taattcttat aatcagcata tgcatgtatt ggtatgtgtg gaaccaactt
     1861 attttaagaa tacagaaaac tacgtgaatc aaaaacaatg gattcaattt tggaaaaagg
     1921 caatgaaatt agactatgat ccaaatgtaa aagttcaaat gattcgaccg aaaaataaat
     1981 ataaatcgga tatacaatcg gcaattgacg aaactgcaaa atatcctgta aaggatacgg
     2041 attttatgac cgatgatgaa gaaaagaatt tgaaacgttt gtctgatttg gaggaaggtt
     2101 tacaccgtaa aaggttaatc tcctatggtg gtttgttaaa agaaatacat aaaaaattaa
     2161 accttgatga cacagaagaa ggcgatttga ttcatacaga tgatgacgaa aaagccgatg
     2221 aagatggatt ttctattatt gcaatgtgga attgggaacg gaaaaattat tttattaaag
     2281 agtagttcaa caaacgggat tgacttttaa aaaaggattg attctaatga agaaagcaga
     2341 caagtaagcc tccgaattcg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     2401 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     2461 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     2521 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     2581 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     2641 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     2701 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     2761 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     2821 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     2881 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     2941 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     3001 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     3061 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     3121 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     3181 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     3241 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     3301 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     3361 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     3421 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     3481 gtgagttttc gttccactga gcgtcaagat ctgttgtgtg gaattgtgag cggataacaa
     3541 tttcacacag gaaacagcta tgaccatgat tacgccaagc ttgcatgcct gcaggtcgac
     3601 tctagaggat cccacctcaa atggttcgct gggtttatcg acctgcagtc acactggctc
     3661 accttcgggt gggcctttct gcgagccctt caaacttccc aaaggcgagc cctagtgaca
     3721 ttagaaaacc gactgtaaaa agtacagtcg gcattatctc atattataaa agccagtcat
     3781 taggcctatc tgacaattcc tgaatagagt tcataaacaa tcctgcatga taaccatcac
     3841 aaacagaatg atgtacctgt aaagatagcg gtaaatatat tgaattacct ttattaatga
     3901 attttcctgc tgtaataatg ggtagaaggt aattactatt attattgata tttaagttaa
     3961 acccagtaaa tgaagtccat ggaataatag aaagagaaaa agcattttca ggtataggtg
     4021 ttttgggaaa caatttcccc gaaccattat atttctctac atcagaaagg tataaatcat
     4081 aaaactcttt gaagtcattc tttacaggag tccaaatacc agagaatgtt ttagatacac
     4141 catcaaaaat tgtataaagt ggctctaact tatcccaata acctaactct ccgtcgctat
     4201 tgtaaccagt tctaaaagct gtatttgagt ttatcaccct tgtcactaag aaaataaatg
     4261 cagggtaaaa tttatatcct tcttgtttta tgtttcggta taaaacacta atatcaattt
     4321 ctgtggttat actaaaagtc gtttgttggt tcaaataatg attaaatatc tcttttctct
     4381 tccaattgtc taaatcaatt ttattaaagt tcatcattaa tatcgcctcc tattgtaaat
     4441 taaaatttaa tttaatgcct ctattagctg ccataaagca tgaatgcagt aaaatattca
     4501 tcctctggca agtatctctt gcactatgat tcattttata ttattttatt ttcattttca
     4561 atagtttaga taaaaaataa ttaatttttt aacaaagtat ttttatcttc ctttgatgta
     4621 attctttgtt tttttgtcca ttaatattat ggacaatttt tttatttcta gtcaataatt
     4681 ttaatcttta ttaaaacctg gcaatgccag agagatacag taattattgc atataaaaga
     4741 aaacagacgc gccctgtttg tttcctgaaa aaccacctcg tcctccaaag tatccagctt
     4801 tttaaaatcc atcctcattc ccagacgaaa aagttttgac aaagcctctt ttcttgtcca
     4861 tagaattgta ctctgcacac aatattcatc agtatttcca aaactcttta gaatcttttc
     4921 ctctctttcg gaaaagaaat acaggaggct tacttgtctg ctttcttcat tagaatcaat
     4981 ccttttttaa aagtcaatcc cgtttgttga actacaccgg ttacaagggt taccggaatt
     5041 tacaataaaa aaggccatcc gtcaggatgg ccttctgctt aatttgatgc ctggcagttt
     5101 atggcgggcg tcctgcccgc caccctccgg gccgttgctt cgcaacgttc aaatccgctc
     5161 ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg
     5221 cccagtcttt cgactgagcc tttcgtttta tcgtttcact tctgagttcg gcatggggtc
     5281 aggtgggacc accgcgctac tgccctcgag gagctcgaat tgaaatatta tttgtataaa
     5341 aaaattacac aaaaaaaatt acataaaaaa aatttgtata ataataatac cgtctgtcgt
     5401 acaatgcgag ggatccccgg gtaccgagct cgaattgaca gatcgctgag ataggtgcct
     5461 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     5521 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5581 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5641 aaggatcttc
//
