LOCUS       V008008                10309 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V008008
VERSION     V008008
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10309)
  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 10309)
  AUTHORS   Olson DG.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-AUG-2012) Thayer School of Engineering, Dartmouth
            College, Thayer School of Engineering, Dartmouth College, Hanover,
            NH 03755, USA
REFERENCE   3  (bases 1 to 10309)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10309)
  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
            (14-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..10309
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    11..514
                     /label="CEN/ARS"
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     misc_feature    680..1214
                     /label="5' flank (CipA + promoter)"
                     /note="5' flank (CipA + promoter)"
     misc_feature    1215..1753
                     /label="3' flank"
                     /note="3' flank"
     regulatory      1766..2413
                     /label="gapD promoter from Clostridium thermocellum"
                     /note="gapD promoter from Clostridium thermocellum"
                     /regulatory_class="promoter"
     CDS             2414..3061
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     CDS             3082..3627
                     /codon_start=1
                     /gene="hpt"
                     /product="putative hypoxanthine phosphoribosyltransferase"
                     /EC_number="2.4.2.8"
                     /label="hpt"
                     /protein_id="AFU51886.1"
                     /translation="MENLSKDIDEILITEEELKEKIKELGRQITKDYKGKNLMLVGVLK
                     GALMFMADLSRHIDLPLSLDFMAVSSYGSSTHSSGIVKIIKDLDISIEGKDVLIVEDII
                     DSGLTLSYLRETLLGRKPKSLKICTILDKPERREASVKVDYVGFKIPDKFVVGYGLDFD
                     EKYRNLPFIGVLKPEMYS"
     gene            3082..3627
                     /gene="hpt"
                     /label="hpt"
     terminator      4305..4552
                     /label="CYC1 terminator"
                     /note="transcription terminator for CYC1"
     rep_origin      complement(4800..5388)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5475..6053)
                     /codon_start=1
                     /gene="tdk"
                     /product="thymidine kinase"
                     /EC_number="2.7.1.21"
                     /label="tdk"
                     /protein_id="AFU51887.1"
                     /translation="MYGPKDHGYIEVVTGPMFSGKSEELIRRIKRAKIARQKVQVFKPA
                     IDDRYSIDKVVSHNGDNMHAIAIVKASDILAYAEEDTDVFAIDEVQFFDSEIVDIVKEI
                     ADSGKRVICAGLDMDFRGEPFGPTPELMAIAEFVDKLTAICMKCGNPATRTQRLINGKP
                     ANYDDPIIMVGAKESYEARCRKCHEVPRT"
     gene            complement(5475..6053)
                     /gene="tdk"
                     /label="tdk"
     regulatory      complement(6054..6674)
                     /note="CBP; cellulose-binding protein promoter"
                     /regulatory_class="promoter"
     CDS             complement(6753..7754)
                     /label="repB"
                     /note="RepB replication protein"
     CDS             complement(8315..9172)
                     /label="AmpR"
                     /note="beta-lactamase"
     CDS             complement(9271..10071)
                     /label="URA3"
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     promoter        complement(10072..10292)
                     /label="URA3 promoter"
ORIGIN
        1 ggtccttttc atcacgtgct ataaaaataa ttataattta aattttttaa tataaatata
       61 taaattaaaa atagaaagta aaaaaagaaa ttaaagaaaa aatagttttt gttttccgaa
      121 gatgtaaaag actctagggg gatcgccaac aaatactacc ttttatcttg ctcttcctgc
      181 tctcaggtat taatgccgaa ttgtttcatc ttgtctgtgt agaagaccac acacgaaaat
      241 cctgtgattt tacattttac ttatcgttaa tcgaatgtat atctatttaa tctgcttttc
      301 ttgtctaata aatatatatg taaagtacgc tttttgttga aattttttaa acctttgttt
      361 attttttttt cttcattccg taactcttct accttcttta tttactttct aaaatccaaa
      421 tacaaaacat aaaaataaat aaacacagag taaattccca aattattcca tcattaaaag
      481 atacgaggcg cgtgtaagtt acaggcaagc gatccgtccg ccggcgaacg tggcgagaaa
      541 ggaagggaag aaagcgaaag gagcgggggc tagggcggtg ggaagtgtag gggtcacgct
      601 gggcgtaacc accacacccg ccgcgcttaa tggggcgcta cagggcgcgt ggggatgatc
      661 cactagtaag cttggatccg gatgaattgt ttactgatga tgaacgaaag aaactgattg
      721 aattgcatat gttatgcaac aaagatgcga atttgccgaa ggtatccgtt tgctcttact
      781 ttgaatcccg ggagatgttc ggatgcggtg ctgcgggata ctattcattt attgatatac
      841 atggtgattt atatccgtgc gattacatac cctttaattt tggcaatgtt tttgaaactc
      901 cgataaaggc tttatggaaa aagatgtata aaagttttga aacgccggaa gtgcattgcc
      961 gttcgctaat ttgtcttggg aaaatggaac gggataaatt tcaacaatat ccgcaaaggg
     1021 tgaaaacttt gccggaaagt tgcaaaacgt gctgcgggca agtcacaccg ggattttaca
     1081 aaatacttgg gggagaaaag taattttgta ataatacttt caaatgaaaa aatattattt
     1141 aaaaaaatgc ccctcaaatt ccgtttatat atcgaatata tattacaaga aagaagatgc
     1201 ccagataaaa agagatatta aaattgggag gaaggatacc ccccggtatc cttcctctca
     1261 aaaatattct ttttttatat ttgaaaagca gaaagagaga aacagattaa aaattagagc
     1321 tatatgtgct atacatgagc tgttgaaggg gggaattttt tcttcatgaa acgaaaaaat
     1381 aaagtattat caattttgtt aactctgctg ctaataatct ctaccacatc cgtaaacatg
     1441 tcttttgctg aagcaactcc aagtattgaa atggttcttg ataaaactga agtccatgta
     1501 ggagatgtaa taacggccac aataaaagtc aataacatta gaaaattggc gggatatcag
     1561 ctaaatatca aatttgaccc tgaagtttta cagccggtag accctgcaac aggagaggaa
     1621 tttactgata agtccatgcc ggtaaatagg gttttgctga caaacagcaa atatggacct
     1681 actcctgtgg cgggtaacga tataaagtca ggaattatta attttgctac gggatataac
     1741 aatttaacag cgttcgcgag gccggccagt attctgacat gggtgtatca ataacccatg
     1801 cgtttccgta ttgtatcgga atggtttcgg acagggcggt gggaatagac atggaaaaga
     1861 tttttttgcc cgaggatgca ttgataaagt atttcttttc cgaaagagag gaaaagattc
     1921 taaagagttt tggaaatact gatgaatatt gtgtgcagag tacaattcta tggacaagaa
     1981 aagaggcttt gtcaaaactt tttcgtctgg gaatgaggat ggattttaaa aagctggata
     2041 ctttggagga cgaggtggtt tttcaggaaa caaacagggc gcgtctgttt tcttttatat
     2101 gcaataatta ctgtatctct ctggcattgc caggttttaa taaagattaa aattattgac
     2161 tagaaataaa aaaattgtcc ataatattaa tggacaaaaa aacaaagaat tacatcaaag
     2221 gaagataaaa atactttgtt aaaaaattaa ttatttttta tctaaactat tgaaaatgaa
     2281 aataaaataa tataaaatga atcatagtgc aagagatact tgccagagga tgaatatttt
     2341 actgcattca tgctttatgg cagctaatag aggcattaaa ttaaatttta atttacaata
     2401 ggaggcgata ttaatgaact ttaataaaat tgatttagac aattggaaga gaaaagagat
     2461 atttaatcat tatttgaacc aacaaacgac ttttagtata accacagaaa ttgatattag
     2521 tgttttatac cgaaacataa aacaagaagg atataaattt taccctgcat ttattttctt
     2581 agtgacaagg gtgataaact caaatacagc ttttagaact ggttacaata gcgacggaga
     2641 gttaggttat tgggataagt tagagccact ttatacaatt tttgatggtg tatctaaaac
     2701 attctctggt atttggactc ctgtaaagaa tgacttcaaa gagttttatg atttatacct
     2761 ttctgatgta gagaaatata atggttcggg gaaattgttt cccaaaacac ctatacctga
     2821 aaatgctttt tctctttcta ttattccatg gacttcattt actgggttta acttaaatat
     2881 caataataat agtaattacc ttctacccat tattacagca ggaaaattca ttaataaagg
     2941 taattcaata tatttaccgc tatctttaca ggtacatcat tctgtttgtg atggttatca
     3001 tgcaggattg tttatgaact ctattcagga attgtcagat aggcctaatg actggctttt
     3061 ataataaagg aggtcgacgt catggaaaat ttatcaaaag acatcgatga aattttgatc
     3121 acagaagaag aacttaagga aaagataaaa gagcttggga ggcaaatcac aaaagactac
     3181 aaagggaaaa atttgatgtt ggtaggagtt ttaaaaggtg ctttaatgtt tatggctgat
     3241 ttgtcaagac acatagattt gcctttatca cttgatttta tggctgtttc cagctatgga
     3301 agctcaactc attcatcagg aatagtaaag ataatcaaag atcttgatat aagcatagaa
     3361 ggcaaagatg ttctgattgt ggaagacata attgacagcg gtttgacttt gtcttactta
     3421 agggaaactt tacttggaag gaagccaaaa agcctgaaaa tatgcacaat attagacaaa
     3481 ccggagagaa gagaagcatc tgtaaaagtc gattatgtag gatttaagat acctgataag
     3541 tttgtcgtgg gttatggatt ggactttgat gaaaagtaca ggaaccttcc ttttataggc
     3601 gttttgaaac ctgaaatgta cagctaagcc cgggcctcga gaaaacaaaa ggctcagtcg
     3661 gaagactggg ccttttgttt tggtacccct gtatatttca gaggagttcc atccaaagga
     3721 atagcaaact gcgactttgt gttcagatat gatccgaatg tattggaaat tatagggata
     3781 gatcccggag acataatagt tgacccgaat cctaccaaga gctttgatac tgcaatatat
     3841 cctgacagaa agataatagt attcctgttt gcggaagaca gcggaacagg agcgtatgca
     3901 ataactaaag acggagtatt tgcaaaaata agagcaactg taaaatcaag tgctccgggc
     3961 tatattactt tcgacgaagt aggtggattt gcagataatg acctggtaga acagaaggta
     4021 tcatttatag acggtggtgt taacgttggc aatgcaacac cgaccaaggg agcaacacca
     4081 acaaatacag ctacgccgac aaaatcagct acggctacgc ccaccaggcc atcggtaccg
     4141 acaaacacac cgacaaacac accggcaaat acaccggtat caggcaattt gaaggttgaa
     4201 ttctacaaca gcaatccttc agatactact aactcaatca atcctcagtt caagggaatt
     4261 cggcgcgcct cagcgtttaa acccgctgat cctagagggc cgcatcatgt aattagttat
     4321 gtcacgctta cattcacgcc ctccccccac atccgctcta accgaaaagg aaggagttag
     4381 acaacctgaa gtctaggtcc ctatttattt ttttatagtt atgttagtat taagaacgtt
     4441 atttatattt caaatttttc ttttttttct gtacagacgc gtgtacgcat gtaacattat
     4501 actgaaaacc ttgcttgaga aggttttggg acgctcgaag gctttaattt gcaagctgcg
     4561 gccctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4621 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4681 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4741 atgtgagcaa aaggccagca aaagcccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4801 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4861 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4921 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4981 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5041 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5101 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5161 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5221 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5281 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5341 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5401 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5461 atgaaaaaat ccgcttaagt ccgcgggact tcatggcact ttctacacct tgcttcataa
     5521 gactcctttg ctccaaccat tataatggga tcgtcgtaat tggcaggctt cccatttata
     5581 agcctttgcg tgcgagtagc aggattgcca cacttcatgc atatagcagt aagcttatcg
     5641 acaaattcag ctatggccat caattctgga gtcggaccaa atggttcacc tctaaagtcc
     5701 atgtcaagcc ctgcacatat gactcttttc ccgctatcgg caatctcttt tacgatgtcg
     5761 actatttcag aatcaaaaaa ttgaacttca tctatagcaa atacatccgt atcttcttca
     5821 gcataagcca atatgtcaga agcctttact atggcaatgg cgtgcatgtt gtcgccgtta
     5881 tgagatacga ctttgtctat ggaataccta tcgtctatag ccggtttgaa aacctgaact
     5941 ttttgcctgg caatcttagc tctctttatc cttcttataa gttcctcact ttttccactg
     6001 aacatgggac cagttacaac ttctatgtac ccgtggtctt taggcccata catctttcgt
     6061 cctccttaaa attttcgttt atttgttttt tatttagtgt gcctaaaagc accatcttat
     6121 acattcgtac cttatatctc ttatcttgta tctcttacct catatctctt attattaact
     6181 accgaagtat attttcataa cggaatctta tgacttattt ttctgactgc cgttttacca
     6241 tacggttaat ttatacggaa acagctaatc taaccttgct gaaggcatgt ttaccaaaat
     6301 attatatcta ctttaccata ttaattcaag tggtcatttt gatattttac gccaaaattt
     6361 ttagcgttcc tttacttgat aatttatcca aaaaccttca ttcgtcaaaa caattatgat
     6421 ataattaatt tatgatatga ttaattaata tttacggata actaatcatt tcaatgcaat
     6481 agatcaacat gcaaatcaat atccacctta ctccgaacat tttgcaccaa cgaatgccca
     6541 gcactatatc ccggcaagca atataacgtc tataattttt tatgataacg ataaaattaa
     6601 attttatgaa aggggtaaaa gcatgagaaa aatagctgta ttgttaatta ctttgacgtt
     6661 cttagtcacg actcgaattt aggaggctta cttgtctgct ttcttcatta gaatcaatcc
     6721 ttttttaaaa gtcaatcccg tttgttgaac tactctttaa taaaataatt tttccgttcc
     6781 caattccaca ttgcaataat agaaaatcca tcttcatcgg ctttttcgtc atcatctgta
     6841 tgaatcaaat cgccttcttc tgtgtcatca aggtttaatt ttttatgtat ttcttttaac
     6901 aaaccaccat aggagattaa ccttttacgg tgtaaacctt cctccaaatc agacaaacgt
     6961 ttcaaattct tttcttcatc atcggtcata aaatccgtat cctttacagg atattttgca
     7021 gtttcgtcaa ttgccgattg tatatccgat ttatatttat ttttcggtcg aatcatttga
     7081 acttttacat ttggatcata gtctaatttc attgcctttt tccaaaattg aatccattgt
     7141 ttttgattca cgtagttttc tgtattctta aaataagttg gttccacaca taccaataca
     7201 tgcatgtgct gattataaga attatcttta ttatttattg tcacttccgt tgcacgcata
     7261 aaaccaacaa gatttttatt aattttttta tattgcatca ttcggcgaaa tccttgagcc
     7321 atatctgaca aactcttatt taattcttcg ccatcataaa catttttaac tgttaatgtg
     7381 agaaacaacc aacgaactgt tggcttttgt ttaataactt cagcaacaac cttttgtgac
     7441 tgaatgccat gtttcattgc tctcctccag ttgcacattg gacaaagcct ggatttacaa
     7501 aaccacactc gatacaactt tctttcgcct gtttcacgat tttgtttata ctctaatatt
     7561 tcagcacaat cttttactct ttcagccttt ttaaattcaa gaatatgcag aagttcaaag
     7621 taatcaacat tagcgatttt cttttctctc catggtctca cttttccact ttttgtcttg
     7681 tccactaaaa cccttgattt ttcatctgaa taaatgctac tattaggaca cataatatta
     7741 aaagaaaccc ccatctattt agttatttgt ttggtcactt ataactttaa cagatggggt
     7801 ttttctgtgc aaccaatttt aagggttttc aatactttaa aacacataca taccaacact
     7861 tcaacgcacc tttcagcaac taaaataaaa atgacgttat ttctatatgt atcaagaata
     7921 gaaagaactc gtttttcgct acgctcaaaa cgcaaaaaaa gcactcattc gagtgctttt
     7981 tcttatcgct ccaaatcatg cgattttttc ctctttgctt ttctttgctc acgaagttct
     8041 cgatcacgct gcaaaacatc ttgaagcgaa aaagtattct tcttttcttc cgatcgctca
     8101 tgctgacgca cgaaaagccc tctaggcgca taggaacaac tcctaaatgc atgtgagggg
     8161 ttttctcgtc catgtgaaca gtcgcatacg caatattttg tttcccatag cggccgcgat
     8221 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     8281 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     8341 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     8401 ctacgatacg ggagcgctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     8461 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     8521 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     8581 taagtagttc gccagttaat agtttgcgca acgttgttgg cattgctaca ggcatcgtgg
     8641 tgtcactctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     8701 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     8761 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     8821 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     8881 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     8941 gtgtatcaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     9001 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     9061 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     9121 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     9181 tttttcaatg ggtaataact gatataatta aattgaagct ctaatttgtg agtttagtat
     9241 acatgcattt acttataata cagtttttta gttttgctgg ccgcatcttc tcaaatatgc
     9301 ttcccagcct gcttttctgt aacgttcacc ctctacctta gcatcccttc cctttgcaaa
     9361 tagtcctctt ccaacaataa taatgtcaga tcctgtagag accacatcat ccacggttct
     9421 atactgttga cccaatgcgt ctcccttgtc atctaaaccc acaccgggtg tcataatcaa
     9481 ccaatcgtaa ccttcatctc ttccacccat gtctctttga gcaataaagc cgataacaaa
     9541 atctttgtcg ctcttcgcaa tgtcaacagt acccttagta tattctccag tagataggga
     9601 gcccttgcat gacaattctg ctaacatcaa aaggcctcta ggttcctttg ttacttcttc
     9661 tgccgcctgc ttcaaaccgc taacaatacc tgggcccacc acaccgtgtg cattcgtaat
     9721 gtctgcccat tctgctattc tgtatacacc cgcagagtac tgcaatttga ctgtattacc
     9781 aatgtcagca aattttctgt cttcgaagag taaaaaattg tacttggcgg ataatgcctt
     9841 tagcggctta actgtgccct ccatggaaaa atcagtcaag atatccacat gtgtttttag
     9901 taaacaaatt ttgggaccta atgcttcaac taactccagt aattccttgg tggtacgaac
     9961 atccaatgaa gcacacaagt ttgtttgctt ttcgtgcatg atattaaata gcttggcagc
    10021 aacaggacta ggatgagtag cagcacgttc cttatatgta gctttcgaca tgatttatct
    10081 tcgtttcctg caggtttttg ttctgtgcag ttgggttaag aatactgggc aatttcatgt
    10141 ttcttcaaca ctacatatgc gtatatatac caatctaagt ctgtgctcct tccttcgttc
    10201 ttccttctgt tcggagatta ccgaatcaaa aaaatttcaa agaaaccgaa atcaaaaaaa
    10261 agaataaaaa aaaaatgatg aattgaattg aaaagctagc ttatcgatg
//
