LOCUS       V009618                 8327 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V009618
VERSION     V009618
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8327)
  AUTHORS   Ghosh IN, Landick R.
  TITLE     OptSSeq: High-throughput sequencing readout of growth enrichment
            defines optimal gene expression elements for homoethanologenesis
  JOURNAL   ACS Synth Biol (2016) In press
   PUBMED   27404024
REFERENCE   2  (bases 1 to 8327)
  AUTHORS   Ghosh IN, Landick RC.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-JUL-2016) DOE Great Lakes Bioenergy Research Center,
            University of Wisconsin-Madison, 1552 University Avenue, Madison, WI
            53726, United States
REFERENCE   3  (bases 1 to 8327)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8327)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth
            Biol (2016) In press"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (15-JUL-2016) DOE Great Lakes Bioenergy Research Center, University
            of Wisconsin-Madison, 1552 University Avenue, Madison, WI 53726,
            United States"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8327
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(161..820)
                     /label="pBBR1 Rep"
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
     rep_origin      complement(821..1590)
                     /direction=LEFT
                     /label="pBBR1 oriV"
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1
                     Rep protein for replication"
     promoter        2968..3059
                     /label="AmpR promoter"
     CDS             3060..3848
                     /label="SmR"
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     regulatory      4048..4095
                     /label="B. subtilis glnA terminator"
                     /note="B. subtilis glnA terminator"
                     /regulatory_class="terminator"
     protein_bind    complement(4107..4126)
                     /label="lac operator (symmetric)"
                     /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). The
                     symmetric lac operator was optimized for tight binding of
                     lac repressor."
     promoter        4154..4183
                     /label="trc promoter"
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     regulatory      4235..4243
                     /label="adhA RBS#10"
                     /note="adhA RBS#10"
                     /regulatory_class="ribosome_binding_site"
     CDS             4249..5259
                     /gene="adhA"
                     /label="Alcohol dehydrogenase 1"
                     /note="Alcohol dehydrogenase 1 from Zymomonas mobilis
                     subsp. mobilis (strain ATCC 31821 / ZM4 / CP4). Accession#:
                     P20368"
     regulatory      5286..5294
                     /label="adhB RBS#1"
                     /note="adhB RBS#1"
                     /regulatory_class="ribosome_binding_site"
     CDS             5300..6448
                     /gene="adhB"
                     /label="Alcohol dehydrogenase 2"
                     /note="Alcohol dehydrogenase 2 from Zymomonas mobilis
                     subsp. mobilis (strain ATCC 31821 / ZM4 / CP4). Accession#:
                     P0DJA2"
     regulatory      6467..6475
                     /label="pdc RBS#4"
                     /note="pdc RBS#4"
                     /regulatory_class="ribosome_binding_site"
     CDS             6481..8184
                     /gene="pdc"
                     /label="Pyruvate decarboxylase"
                     /note="Pyruvate decarboxylase from Zymomonas mobilis subsp.
                     mobilis (strain ATCC 31821 / ZM4 / CP4). Accession#:
                     P06672"
     regulatory      8231..8282
                     /label="Phage P22 terminator"
                     /note="Phage P22 terminator"
                     /regulatory_class="terminator"
ORIGIN
        1 cgttagcgag gtgccgccgg cttccattca ggtcgaggtg gcccggctcc atgcaccgcg
       61 acgcaacgcg gggaggcaga caaggtatag ggcggcgagg cggctacagc cgatagtctg
      121 gaacagcgca cttacgggtt gctgcgcaac ccaagtgcta ccggcgcggc agcgtgaccc
      181 gtgtcggcgg ctccaacggc tcgccatcgt ccagaaaaca cggctcatcg ggcatcggca
      241 ggcgctgctg cccgcgccgt tcccattcct ccgtttcggt caaggctggc aggtctggtt
      301 ccatgcccgg aatgccgggc tggctgggcg gctcctcgcc ggggccggtc ggtagttgct
      361 gctcgcccgg atacagggtc gggatgcggc gcaggtcgcc atgccccaac agcgattcgt
      421 cctggtcgtc gtgatcaacc accacggcgg cactgaacac cgacaggcgc aactggtcgc
      481 ggggctggcc ccacgccacg cggtcattga ccacgtaggc cgacacggtg ccggggccgt
      541 tgagcttcac gacggagatc cagcgctcgg ccaccaagtc cttgactgcg tattggaccg
      601 tccgcaaaga acgtccgatg agcttggaaa gtgtcttctg gctgaccacc acggcgttct
      661 ggtggcccat ctgcgccacg aggtgatgca gcagcattgc cgccgtgggt ttcctcgcaa
      721 taagcccggc ccacgcctca tgcgctttgc gttccgtttg cacccagtga ccgggcttgt
      781 tcttggcttg aatgccgatt tctctggact gcgtggccat gcttatctcc atgcggtagg
      841 gtgccgcacg gttgcggcac catgcgcaat cagctgcaac ttttcggcag cgcgacaaca
      901 attatgcgtt gcgtaaaagt ggcagtcaat tacagatttt ctttaaccta cgcaatgagc
      961 tattgcgggg ggtgccgcaa tgagctgttg cgtacccccc ttttttaagt tgttgatttt
     1021 taagtctttc gcatttcgcc ctatatctag ttctttggtg cccaaagaag ggcacccctg
     1081 cggggttccc ccacgccttc ggcgcggctc cccctccggc aaaaagtggc ccctccgggg
     1141 cttgttgatc gactgcgcgg ccttcggcct tgcccaaggt ggcgctgccc ccttggaacc
     1201 cccgcactcg ccgccgtgag gctcgggggg caggcgggcg ggcttcgcct tcgactgccc
     1261 ccactcgcat aggcttgggt cgttccaggc gcgtcaaggc caagccgctg cgcggtcgct
     1321 gcgcgagcct tgacccgcct tccacttggt gtccaaccgg caagcgaagc gcgcaggccg
     1381 caggccggag gcttttcccc agagaaaatt aaaaaaattg atggggcaag gccgcaggcc
     1441 gcgcagttgg agccggtggg tatgtggtcg aaggctgggt agccggtggg caatccctgt
     1501 ggtcaagctc gtgggcaggc gcagcctgtc catcagcttg tccagcaggg ttgtccacgg
     1561 gccgagcgaa gcgagccagc cggtggccgc tcgcggccat cgtccacata tccacgggct
     1621 ggcaagggag cgcagcgacc gcgcagggcg aagcccggag agcaagcccg tagggcgccg
     1681 cagccgccgt aggcggtcac gactttgcga agcaaagtct agtgagtata ctcaagcatt
     1741 gagtggcccg ccggaggcac cgccttgcgc tgcccccgtc gagccggttg gacaccaaaa
     1801 gggaggggca ggcatggcgg catacgcgat catgcgatgc aagaagctgg cgaaaatggg
     1861 caacgtggcg gccagtctca agcacgccta ccgcgagcgc gagacgccca acgctgacgc
     1921 cagcaggacg ccagagaacg agcactgggc ggccagcagc accgatgaag cgatgggccg
     1981 actgcgcgag ttgctgccag agaagcggcg caaggacgct gtgttggcgg tcgagtacgt
     2041 catgacggcc agcccggaat ggtggaagtc ggccagccaa gaacagcagg cggcgttctt
     2101 cgagaaggcg cacaagtggc tggcggacaa gtacggggcg gatcgcatcg tgacggccag
     2161 catccaccgt gacgaaacca gcccgcacat gaccgcgttc gtggtgccgc tgacgcagga
     2221 cggcaggctg tcggccaagg agttcatcgg caacaaagcg cagatgaccc gcgaccagac
     2281 cacgtttgcg gccgctgtgg ccgatctagg gctgcaacgg ggcatcgagg gcagcaaggc
     2341 acgtcacacg cgcattcagg cgttctacga ggccctggag cggccaccag tgggccacgt
     2401 caccatcagc ccgcaagcgg tcgagccacg cgcctatgca ccgcagggat tggccgaaaa
     2461 gctgggaatc tcaaagcgcg ttgagacgcc ggaagccgtg gccgaccggc tgacaaaagc
     2521 ggttcggcag gggtatgagc ctgccctaca ggccgccgca ggagcgcgtg agatgcgcaa
     2581 gaaggccgat caagcccaag agacggcccg agaccttcgg gagcgcctga agcccgttct
     2641 ggacgccctg gggccgttga atcgggatat gcaggccaag gccgccgcga tcatcaaggc
     2701 cgtgggcgaa aagctgctga cggaacagcg ggaagtccag cgccagaaac aggcccagcg
     2761 ccagcaggaa cgcgggcgcg cacatttccc cgaaaagtgc cacctggcgg cgttgtgaca
     2821 atttaccgaa caactccgcg gccgggaagc cgatctcggc ttgaacgaat tcctgcttat
     2881 ccacggaatc tgtgggtaac tttgtatgtg tccgcagcgc ccgccgcagt ctcacgcccg
     2941 gagcgtagcg accgagtgag ctagctattt gtttattttt ctaaatacat tcaaatatgt
     3001 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     3061 tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg
     3121 agcgccatct cgaaccgacg ttgctggccg tacatttgta cggctccgca gtggatggcg
     3181 gcctgaagcc acacagtgat attgatttgc tggttacggt gaccgtaagg cttgatgaaa
     3241 caacgcggcg agctttgatc aacgaccttt tggaaacttc ggcttcccct ggagagagcg
     3301 agattctccg cgctgtagaa gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt
     3361 atccagctaa gcgcgaactg caatttggag aatggcagcg caatgacatt cttgcaggta
     3421 tcttcgagcc agccacgatc gacattgatc tggctatctt gctgacaaaa gcaagagaac
     3481 atagcgttgc cttggtaggt ccagcggcgg aggaactctt tgatccggtt cctgaacagg
     3541 atctatttga ggcgctaaat gaaaccttaa cgctatggaa ctcgccgccc gactgggctg
     3601 gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg gtacagcgca gtaaccggca
     3661 aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga gcgcctgccg gcccagtatc
     3721 agcccgtcat acttgaagct agacaggctt atcttggaca agaagaagat cgcttggcct
     3781 cgcgcgcaga tcagttggaa gaatttgtcc actacgtgaa aggcgagatc accaaggtag
     3841 tcggcaaata atgtctaaca attcgttcaa gccgaggggc cgcaagatcc ggccacgatg
     3901 acccggtcgt cggttcaggg cagggtcgtt aaatagccgc ttatgtctat tgctggttta
     3961 ccggtttatt gactaccgga agcagtgtga ccgtgtgctt cttactacgt cagtgtcgtg
     4021 ccatagactg gaaagcgggc agtaactact caatcccttg gcactaaaag tgtcagggga
     4081 ttttttatgt taatacacca tcacagaatt gtgagcgctc acaatctagg tctatgagtg
     4141 gttgctggac ctcttgacaa ttaatcatcc ggctcggata atgggtggaa ttggtagagt
     4201 atttttattg cgcggtcacc cgccatatat catctaaaga tgttcaccat gaaagctgca
     4261 gttatcacta aagatcatac gatcgaagtg aaagacacca aattacgccc gctgaaatat
     4321 ggcgaggctc tgctggagat ggaatattgc ggtgtctgtc acacggatct gcatgtcaaa
     4381 aacggtgact ttggtgatga aacgggtcgc atcaccggcc acgagggtat tggtatcgtg
     4441 aaacaggttg gtgagggcgt tacttctctg aaagtcggtg atcgtgccag tgttgcatgg
     4501 ttcttcaaag gctgcggcca ttgtgaatac tgtgttagcg gtaacgaaac cctgtgccgt
     4561 aatgttgaga acgcgggtta tacggtggat ggcgcgatgg cggaagagtg cattgtggtc
     4621 gcggactata gcgtcaaggt cccggatggc ctggacccag ccgttgcgag cagcatcacc
     4681 tgcgcgggtg taaccaccta taaagcagtc aaagtttctc agatacagcc gggacaatgg
     4741 ctggctatct atggcttggg cggtttaggc aatctggccc tgcaatatgc caagaatgtt
     4801 ttcaacgcca aagtcatcgc gattgatgtc aacgacgaac aactggcctt cgccaaggaa
     4861 ttgggtgcgg atatggttat caacccgaaa aacgaggatg ccgcgaagat catccaagag
     4921 aaagtgggtg gtgcgcacgc aaccgtggtg acggcagttg ccaaaagcgc gttcaatagc
     4981 gcagtggagg ctatccgcgc gggtggtcgt gtcgtggcag taggcctgcc gccggagaag
     5041 atggacttat cgattccgcg cctggtgctg gatggcatcg aagttttggg tagcctggtt
     5101 ggtacgcgtg aggatctgaa agaggcgttc caattcgcag ctgagggtaa ggtgaaaccg
     5161 aaagtgacga agcgtaaggt cgaagaaatc aaccaaattt ttgacgagat ggagcacggc
     5221 aagttcacgg gccgtatggt tgtcgacttc acgcatcatt aactttttgc aatctagtat
     5281 ggccttaagg gggatagcta tggcctcgag cacgttctat attccgttcg tgaacgagat
     5341 gggcgaaggc agcttggaga aggctattaa agacctgaac ggtagcggtt tcaaaaacgc
     5401 actgatcgtt agcgatgcct ttatgaacaa gagcggcgtg gtcaagcagg tggcagacct
     5461 gctgaaggcg caaggtatca attcggccgt ttatgacggt gttatgccga acccgaccgt
     5521 tacggcagtt ctggaaggcc tgaagatcct gaaggataac aatagcgact tcgtcatctc
     5581 cctcggcggt ggtagcccac acgattgcgc aaaggcgatc gcgctggtcg cgacgaacgg
     5641 tggcgaggtg aaggactatg aaggtatcga caaatctaag aaacctgccc tgcctttgat
     5701 gagcatcaac acgacggctg gtaccgctag cgaaatgacc cgtttctgca tcatcactga
     5761 tgaagtccgt cacgttaaga tggccattgt tgaccgtcac gttaccccga tggtgtcggt
     5821 aaatgatccg ctgttgatgg tgggtatgcc taagggtctg accgccgcta cgggtatgga
     5881 tgcgctgact cacgcatttg aggcgtacag ctccacggcg gcaaccccta tcaccgacgc
     5941 ctgtgcactg aaagccgcca gcatgatcgc gaagaacctg aaaactgcct gcgacaatgg
     6001 taaagatatg ccggcacgtg aggcgatggc gtatgcgcag ttcctggctg gtatggcgtt
     6061 caataacgcg agcctgggtt atgtgcatgc gatggcccat cagctgggtg gttactataa
     6121 tctgccgcac ggcgtgtgta acgctgtcct gctgccgcat gtgttagcct ataatgcttc
     6181 cgtggtcgca ggccgcctga aggatgtggg tgttgcgatg ggtctggata ttgccaatct
     6241 gggtgacaag gagggcgccg aggcaacgat ccaggccgta cgtgacttgg cagcatccat
     6301 tggcatcccg gctaacctga cggaactggg cgcgaagaaa gaggatgtgc cgcttcttgc
     6361 tgaccacgct ctgaaagatg cttgtgctct gaccaacccg cgtcagggtg atcagaagga
     6421 ggtcgaggag ctgttcctga gcgcattcta attaaatcca ctatcttcag gagatgttac
     6481 atgtcctata ctgtcggtac ctatttagcg gagcgtctgg tccagattgg tctgaagcat
     6541 cacttcgcag tcgcgggcga ctacaacctc gtcctgctgg acaacctgct gctgaacaag
     6601 aacatggagc aagtgtactg ctgtaacgaa ctcaattgcg gttttagcgc agagggttac
     6661 gcacgcgcga aaggcgcggc ggctgcggtt gtcacttatt ctgtcggcgc cctgagcgct
     6721 tttgatgcta tcggtggcgc ctatgcagaa aaccttccgg ttatcctgat ctccggtgct
     6781 ccgaacaata acgatcatgc cgcaggtcac gtcctgcatc acgcgctggg caagacggac
     6841 taccactacc agttggagat ggctaagaat atcactgcgg cggcggaggc tatctacacc
     6901 ccggaggagg caccggcgaa gattgaccac gtgatcaaga ccgcactgcg tgagaagaag
     6961 ccggtttatc tcgaaatcgc ttgcaacatt gcttccatgc cgtgcgccgc accaggtccg
     7021 gcaagcgcgt tattcaacga cgaggcgagc gatgaagcaa gcctgaacgc tgctgtggaa
     7081 gaaacgctca agtttatcgc taaccgcgat aaggttgcag tgttggtcgg ttcgaaactg
     7141 cgtgccgcag gcgcagagga agccgccgta aaattcgcgg atgcattggg tggcgctgtt
     7201 gcaacgatgg cggcagcgaa gagctttttc ccggaagaaa acccacatta catcggcacg
     7261 tcgtggggtg aagtcagcta cccgggtgtt gagaagacga tgaaggaggc cgacgcggtt
     7321 attgccctgg ctccagtctt caatgattac tcgacgaccg gctggaccga tattccggac
     7381 ccaaaaaagc tggtcctggc agaaccgcgt agcgttgtgg tgaatggtgt gcgttttcct
     7441 agcgtccatc tgaaagatta cctgacccgc ctggcccaaa aagtcagcaa gaaaaccggt
     7501 gcgctggact ttttcaaatc actgaacgcg ggtgaattga agaaagcggc ccctgcggat
     7561 ccgagcgcac cgctggttaa cgctgaaatt gctcgccagg tggaagcatt gctgaccccg
     7621 aatacgaccg tgattgctga aaccggtgac tcttggttca atgctcagcg catgaagctc
     7681 ccgaacggtg ctcgcgtgga atacgagatg cagtggggcc atatcggctg gtctgtcccg
     7741 gcagcgttcg gttatgccgt cggtgccccg gaacgtcgca acatcctcat ggttggtgat
     7801 ggttccttcc agttgaccgc gcaggaagtg gctcagatgg tgcgcctgaa actgccggtg
     7861 atcatctttc tgattaacaa ttatggttat accattgaag tgatgatcca cgacggtcct
     7921 tataacaaca tcaaaaactg ggattacgcg ggtttgatgg aagtgttcaa tggcaacggt
     7981 ggttatgaca gcggcgcagg taagggtctg aaagctaaga ccggtggtga gctggctgag
     8041 gcgatcaagg tggccctggc gaacactgat ggcccaacgt tgatcgagtg cttcatcggt
     8101 cgtgaagact gcactgaaga attggtcaaa tggggtaagc gcgttgctgc cgcgaattct
     8161 cgtaagccgg tcaataagtt gctgtaaatc gaattcctgc agcccggggg atccctagtt
     8221 ctagaaataa gttaaataaa gccctgagtt taaccgctcg gggctttttg cgttttaagc
     8281 acactagagc ggccatgcat cccagagcct gataaaaacg gtgaatc
//
