LOCUS       V008009                 9045 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V008009
VERSION     V008009
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9045)
  AUTHORS   Waller BH, Olson DG, Currie DH, Guss AM, Lynd LR.
  TITLE     Exchange of type II dockerin-containing subunits of the Clostridium
            thermocellum cellulosome as revealed by SNAP-tags
  JOURNAL   FEMS Microbiol. Lett. 338 (1), 46-53 (2013)
   PUBMED   23082914
REFERENCE   2  (bases 1 to 9045)
  AUTHORS   Olson DG.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-AUG-2012) Thayer School of Engineering, Dartmouth
            College, Thayer School of Engineering, Dartmouth College, Hanover,
            NH 03755, USA
REFERENCE   3  (bases 1 to 9045)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9045)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "FEMS
            Microbiol. Lett."; date: "2013"; volume: "338"; issue: "1"; pages:
            "46-53"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (13-AUG-2012) Thayer School of Engineering, Dartmouth College,
            Thayer School of Engineering, Dartmouth College, Hanover, NH 03755,
            USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9045
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..589)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(906..1706)
                     /label="URA3"
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     promoter        complement(1707..1927)
                     /label="URA3 promoter"
     misc_feature    1955..2458
                     /label="CEN/ARS"
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     misc_feature    complement(2503..3041)
                     /label="3' flank"
                     /note="3' flank"
     misc_feature    complement(3042..3583)
                     /label="5' flank"
                     /note="5' flank"
     regulatory      complement(3589..3720)
                     /label="hpt"
                     /note="hpt"
                     /regulatory_class="terminator"
     CDS             complement(3721..4275)
                     /codon_start=1
                     /gene="hpt"
                     /product="hypoxanthine phosphoribosyltransferase"
                     /label="hpt"
                     /protein_id="AFU51881.1"
                     /translation="MINQIKEILVTREELKNNAKELGKRISSDYEGKELVLIGVLKGGV
                     VFFADLIREITIPIDVDFISVSSYGNSTKSSGVVRIIKDIDIDITNKHVLIVEDLVDTG
                     LTLHYLKSMFEARGPKDVKICTALDKPSRRKVDLEIDYKGITIPDKFVVGYGLDYAEKY
                     RNLPDVCVLDSSVYTDKEDMD"
     gene            complement(3721..4275)
                     /gene="hpt"
                     /label="hpt"
     CDS             complement(4294..4941)
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     regulatory      complement(4942..5469)
                     /label="gapDH promoter region"
                     /note="gapDH promoter region"
                     /regulatory_class="promoter"
     misc_feature    complement(5473..6040)
                     /label="internal flank"
                     /note="internal flank"
     CDS             complement(6044..6622)
                     /codon_start=1
                     /gene="tdk"
                     /product="thymidine kinase"
                     /label="tdk"
                     /note="from Thermoanaerobacterium saccharolyticum"
                     /protein_id="AFU51883.1"
                     /translation="MYGPKDHGYIEVVTGPMFSGKSEELIRRIKRAKIARQKVQVFKPA
                     IDDRYSIDKVVSHNGDNMHAIAIVKASDILAYAEEDTDVFAIDEVQFFDSEIVDIVKEI
                     ADSGKRVICAGLDMDFRGEPFGPTPELMAIAEFVDKLTAICMKCGNPATRTQRLINGKP
                     ANYDDPIIMVGAKESYEARCRKCHEVPRT"
     gene            complement(6044..6622)
                     /gene="tdk"
                     /label="tdk"
     regulatory      complement(6623..7243)
                     /note="CBP; cellulose-binding protein promoter"
                     /regulatory_class="promoter"
     CDS             complement(7353..8354)
                     /label="repB"
                     /note="RepB replication protein"
ORIGIN
        1 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
       61 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
      121 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
      181 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
      241 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
      301 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
      361 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
      421 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
      481 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
      541 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
      601 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
      661 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
      721 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
      781 ggcacctatc tcagcgatct gtcaattcgc ggccgccgaa aagtgggtaa taactgatat
      841 aattaaattg aagctctaat ttgtgagttt agtatacatg catttactta taatacagtt
      901 ttttagtttt gctggccgca tcttctcaaa tatgcttccc agcctgcttt tctgtaacgt
      961 tcaccctcta ccttagcatc ccttcccttt gcaaatagtc ctcttccaac aataataatg
     1021 tcagatcctg tagagaccac atcatccacg gttctatact gttgacccaa tgcgtctccc
     1081 ttgtcatcta aacccacacc gggtgtcata atcaaccaat cgtaaccttc atctcttcca
     1141 cccatgtctc tttgagcaat aaagccgata acaaaatctt tgtcgctctt cgcaatgtca
     1201 acagtaccct tagtatattc tccagtagat agggagccct tgcatgacaa ttctgctaac
     1261 atcaaaaggc ctctaggttc ctttgttact tcttctgccg cctgcttcaa accgctaaca
     1321 atacctgggc ccaccacacc gtgtgcattc gtaatgtctg cccattctgc tattctgtat
     1381 acacccgcag agtactgcaa tttgactgta ttaccaatgt cagcaaattt tctgtcttcg
     1441 aagagtaaaa aattgtactt ggcggataat gcctttagcg gcttaactgt gccctccatg
     1501 gaaaaatcag tcaagatatc cacatgtgtt tttagtaaac aaattttggg acctaatgct
     1561 tcaactaact ccagtaattc cttggtggta cgaacatcca atgaagcaca caagtttgtt
     1621 tgcttttcgt gcatgatatt aaatagcttg gcagcaacag gactaggatg agtagcagca
     1681 cgttccttat atgtagcttt cgacatgatt tatcttcgtt tcctgcaggt ttttgttctg
     1741 tgcagttggg ttaagaatac tgggcaattt catgtttctt caacactaca tatgcgtata
     1801 tataccaatc taagtctgtg ctccttcctt cgttcttcct tctgttcgga gattaccgaa
     1861 tcaaaaaaat ttcaaagaaa ccgaaatcaa aaaaaagaat aaaaaaaaaa tgatgaattg
     1921 aattgaaaag ctagcttatc gatgggtcct tttcatcacg tgctataaaa ataattataa
     1981 tttaaatttt ttaatataaa tatataaatt aaaaatagaa agtaaaaaaa gaaattaaag
     2041 aaaaaatagt ttttgttttc cgaagatgta aaagactcta gggggatcgc caacaaatac
     2101 taccttttat cttgctcttc ctgctctcag gtattaatgc cgaattgttt catcttgtct
     2161 gtgtagaaga ccacacacga aaatcctgtg attttacatt ttacttatcg ttaatcgaat
     2221 gtatatctat ttaatctgct tttcttgtct aataaatata tatgtaaagt acgctttttg
     2281 ttgaaatttt ttaaaccttt gtttattttt ttttcttcat tccgtaactc ttctaccttc
     2341 tttatttact ttctaaaatc caaatacaaa acataaaaat aaataaacac agagtaaatt
     2401 cccaaattat tccatcatta aaagatacga ggcgcgtgta agttacaggc aagcgatcgc
     2461 ggccgcggta cccggggatc ctctagagtc gacctgcagt gcacgctgtt aaattgttat
     2521 atcccgtagc aaaattaata attcctgact ttatatcgtt acccgccaca ggagtaggtc
     2581 catatttgct gtttgtcagc aaaaccctat ttaccggcat ggacttatca gtaaattcct
     2641 ctcctgttgc agggtctacc ggctgtaaaa cttcagggtc aaatttgata tttagctgat
     2701 atcccgccaa ttttctaatg ttattgactt ttattgtggc cgttattaca tctcctacat
     2761 ggacttcagt tttatcaaga accatttcaa tacttggagt tgcttcagca aaagacatgt
     2821 ttacggatgt ggtagagatt attagcagca gagttaacaa aattgataat actttatttt
     2881 ttcgtttcat gaagaaaaaa ttcccccctt caacagctca tgtatagcac atatagctct
     2941 aatttttaat ctgtttctct ctttctgctt ttcaaatata aaaaaagaat atttttgaga
     3001 ggaaggatac cggggggtat ccttcctccc aattttaata tctaccattc ctcccaaaaa
     3061 ttaatatttt atatgatctt ataaaaagat ctctaaaggc atttgaaatc aaatacctta
     3121 tatacaaacc aagcaagaca ataacaaaaa ttagtgttaa tattaataaa caatattaat
     3181 attttcttgt ccgattttca gttttaataa ataccagaga atttgtcgat aaatattggg
     3241 gagtggtttt taataaatcg atggaacata cgattctatg tatatgcttt gatatttgca
     3301 caatcaatac atataaagta ttgaatacgc tatttcattg aaatgctggg aattaataaa
     3361 cgctctccag tatggttatc ttatgctcaa agaaccaaaa cataaacaga cgtgaccaat
     3421 cccttgtaca aactcctccc tctaaaactt tttatctata taaactctgt atcttgggat
     3481 atgttagagg ccattaaaac aggatttaaa taaaatatta attactgcta catgaactga
     3541 atactgagat attaagcgct taatttaatg aagatttgcc tccgcagctt caactagttc
     3601 cctcctcatt ctttatagca atatagcaaa acactatctc tcatacatat tattataaaa
     3661 tatatattac attatattaa cataacggta ttaaaaacac aagaaacctt tgtatatttt
     3721 ttagtccata tcttctttgt ccgtataaac agacgaatcc agcacgcaca catccgggag
     3781 atttctgtat ttttccgcat aatccaatcc atagcccacc acaaacttat ccggtattgt
     3841 gataccttta taatctattt ccaaatcaac ctttctcctt gacggtttgt caagggcggt
     3901 gcatattttt acatctttgg gtcctctggc ttcaaacatg cttttcagat aatgcagcgt
     3961 aagacctgta tccaccaagt cttcaacgat aaggacatgc ttgttggtta tatctatatc
     4021 gatgtctttt attatacgca caacccccga tgatttggtg gaattgccgt aacttgacac
     4081 cgatatgaaa tccacatcaa tgggtatggt tatttccctt attaagtcgg caaaaaatac
     4141 cactcctcct tttaacaccc ctatcaggac aagctctttt ccttcatagt cactggaaat
     4201 cctctttccc aactctttag cgttgttttt aagttcctct ctggtaacca aaatttcttt
     4261 aatttgattt atcatacatc cttcctccct ttataaaagc cagtcattag gcctatctga
     4321 caattcctga atagagttca taaacaatcc tgcatgataa ccatcacaaa cagaatgatg
     4381 tacctgtaaa gatagcggta aatatattga attaccttta ttaatgaatt ttcctgctgt
     4441 aataatgggt agaaggtaat tactattatt attgatattt aagttaaacc cagtaaatga
     4501 agtccatgga ataatagaaa gagaaaaagc attttcaggt ataggtgttt tgggaaacaa
     4561 tttccccgaa ccattatatt tctctacatc agaaaggtat aaatcataaa actctttgaa
     4621 gtcattcttt acaggagtcc aaataccaga gaatgtttta gatacaccat caaaaattgt
     4681 ataaagtggc tctaacttat cccaataacc taactctccg tcgctattgt aaccagttct
     4741 aaaagctgta tttgagttta tcacccttgt cactaagaaa ataaatgcag ggtaaaattt
     4801 atatccttct tgttttatgt ttcggtataa aacactaata tcaatttctg tggttatact
     4861 aaaagtcgtt tgttggttca aataatgatt aaatatctct tttctcttcc aattgtctaa
     4921 atcaatttta ttaaagttca tcattaatat cgcctcctat tgtaaattaa aatttaattt
     4981 aatgcctcta ttagctgcca taaagcatga atgcagtaaa atattcatcc tctggcaagt
     5041 atctcttgca ctatgattca ttttatatta ttttattttc attttcaata gtttagataa
     5101 aaaataatta attttttaac aaagtatttt tatcttcctt tgatgtaatt ctttgttttt
     5161 ttgtccatta atattatgga caattttttt atttctagtc aataatttta atctttatta
     5221 aaacctggca atgccagaga gatacagtaa ttattgcata taaaagaaaa cagacgcgcc
     5281 ctgtttgttt cctgaaaaac cacctcgtcc tccaaagtat ccagcttttt aaaatccatc
     5341 ctcattccca gacgaaaaag ttttgacaaa gcctcttttc ttgtccatag aattgtactc
     5401 tgcacacaat attcatcagt atttccaaaa ctctttagaa tcttttcctc tctttcggaa
     5461 aagaaatact gcccttgaac tgaggattga ttgagttagt agtatctgaa ggattgctgt
     5521 tgtagaattc aaccttcaaa ttgcctgata ccggtgtatt tgccggtgtg tttgtcggtg
     5581 tgtttgtcgg taccgatggc ctggtgggcg tagccgtagc tgattttgtc ggcgtagctg
     5641 tatttgttgg tgttgctccc ttggtcggtg ttgcattgcc aacgttaaca ccaccgtcta
     5701 taaatgatac cttctgttct accaggtcat tatctgcaaa tccacctact tcgtcgaaag
     5761 taatatagcc cggagcactt gattttacag ttgctcttat ttttgcaaat actccgtctt
     5821 tagttattgc atacgctcct gttccgctgt cttccgcaaa caggaatact attatctttc
     5881 tgtcaggata tattgcagta tcaaagctct tggtaggatt cgggtcaact attatgtctc
     5941 cgggatctat ccctataatt tccaatacat tcggatcata tctgaacaca aagtcgcagt
     6001 ttgctattcc tttggatgga actcctctga aatatacagg gcattaagtc cgcgggactt
     6061 catggcactt tctacacctt gcttcataag actcctttgc tccaaccatt ataatgggat
     6121 cgtcgtaatt ggcaggcttc ccatttataa gcctttgcgt gcgagtagca ggattgccac
     6181 acttcatgca tatagcagta agcttatcga caaattcagc tatggccatc aattctggag
     6241 tcggaccaaa tggttcacct ctaaagtcca tgtcaagccc tgcacatatg actcttttcc
     6301 cgctatcggc aatctctttt acgatgtcga ctatttcaga atcaaaaaat tgaacttcat
     6361 ctatagcaaa tacatccgta tcttcttcag cataagccaa tatgtcagaa gcctttacta
     6421 tggcaatggc gtgcatgttg tcgccgttat gagatacgac tttgtctatg gaatacctat
     6481 cgtctatagc cggtttgaaa acctgaactt tttgcctggc aatcttagct ctctttatcc
     6541 ttcttataag ttcctcactt tttccactga acatgggacc agttacaact tctatgtacc
     6601 cgtggtcttt aggcccatac atctttcgtc ctccttaaaa ttttcgttta tttgtttttt
     6661 atttagtgtg cctaaaagca ccatcttata cattcgtacc ttatatctct tatcttgtat
     6721 ctcttacctc atatctctta ttattaacta ccgaagtata ttttcataac ggaatcttat
     6781 gacttatttt tctgactgcc gttttaccat acggttaatt tatacggaaa cagctaatct
     6841 aaccttgctg aaggcatgtt taccaaaata ttatatctac tttaccatat taattcaagt
     6901 ggtcattttg atattttacg ccaaaatttt tagcgttcct ttacttgata atttatccaa
     6961 aaaccttcat tcgtcaaaac aattatgata taattaattt atgatatgat taattaatat
     7021 ttacggataa ctaatcattt caatgcaata gatcaacatg caaatcaata tccaccttac
     7081 tccgaacatt ttgcaccaac gaatgcccag cactatatcc cggcaagcaa tataacgtct
     7141 ataatttttt atgataacga taaaattaaa ttttatgaaa ggggtaaaag catgagaaaa
     7201 atagctgtat tgttaattac tttgacgttc ttagtcacga ctcttgatat gcctcctaaa
     7261 tttttatcta aagtgaattt aggaggctta cttgtctgct ttcttcatta gaatcaatcc
     7321 ttttttaaaa gtcaatcccg tttgttgaac tactctttaa taaaataatt tttccgttcc
     7381 caattccaca ttgcaataat agaaaatcca tcttcatcgg ctttttcgtc atcatctgta
     7441 tgaatcaaat cgccttcttc tgtgtcatca aggtttaatt ttttatgtat ttcttttaac
     7501 aaaccaccat aggagattaa ccttttacgg tgtaaacctt cctccaaatc agacaaacgt
     7561 ttcaaattct tttcttcatc atcggtcata aaatccgtat cctttacagg atattttgca
     7621 gtttcgtcaa ttgccgattg tatatccgat ttatatttat ttttcggtcg aatcatttga
     7681 acttttacat ttggatcata gtctaatttc attgcctttt tccaaaattg aatccattgt
     7741 ttttgattca cgtagttttc tgtattctta aaataagttg gttccacaca taccaataca
     7801 tgcatgtgct gattataaga attatcttta ttatttattg tcacttccgt tgcacgcata
     7861 aaaccaacaa gatttttatt aattttttta tattgcatca ttcggcgaaa tccttgagcc
     7921 atatctgaca aactcttatt taattcttcg ccatcataaa catttttaac tgttaatgtg
     7981 agaaacaacc aacgaactgt tggcttttgt ttaataactt cagcaacaac cttttgtgac
     8041 tgaatgccat gtttcattgc tctcctccag ttgcacattg gacaaagcct ggatttacaa
     8101 aaccacactc gatacaactt tctttcgcct gtttcacgat tttgtttata ctctaatatt
     8161 tcagcacaat cttttactct ttcagccttt ttaaattcaa gaatatgcag aagttcaaag
     8221 taatcaacat tagcgatttt cttttctctc catggtctca cttttccact ttttgtcttg
     8281 tccactaaaa cccttgattt ttcatctgaa taaatgctac tattaggaca cataatatta
     8341 aaagaaaccc ccatctattt agttatttgt ttggtcactt ataactttaa cagatggggt
     8401 ttttctgtgc aaccaatttt aagggttttc aatactttaa aacacataca taccaacact
     8461 tcaacgcacc tttcagcaac taaaataaaa atgacgttat ttctatatgt atcaagaata
     8521 gaaagaactc gtttttcgct acgctcaaaa cgcaaaaaaa gcactcattc gagtgctttt
     8581 tcttatcgct ccaaatcatg cgattttttc ctctttgctt ttctttgctc acgaagttct
     8641 cgatcacgct gcaaaacatc ttgaagcgaa aaagtattct tcttttcttc cgatcgctca
     8701 tgctgacgca cgaaaagccc tctaggcgca taggaacaac tcctaaatgc atgtgagggg
     8761 ttttctcgtc catgtgaaca gtcgcatacg caatattttg tttcccatac tgcattaatg
     8821 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     8881 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     8941 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     9001 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtt
//
