LOCUS       V004172                10456 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004172
VERSION     V004172
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10456)
  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 10456)
  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 10456)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10456)
  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..10456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      35..96
                     /function="Stationary phase promoter"
                     /label="Dps100"
                     /note="Dps100"
                     /regulatory_class="promoter"
     regulatory      108..181
                     /function="Expressed in anaerobic conditions"
                     /label="YdfZ promoter"
                     /note="YdfZ promoter"
                     /regulatory_class="promoter"
     CDS             221..1924
                     /gene="pdc"
                     /label="Pyruvate decarboxylase"
                     /note="Pyruvate decarboxylase from Zymomonas mobilis subsp.
                     mobilis (strain ATCC 31821 / ZM4 / CP4). Accession#:
                     P06672"
     CDS             1987..3135
                     /gene="adhB"
                     /label="Alcohol dehydrogenase 2"
                     /note="Alcohol dehydrogenase 2 from Zymomonas mobilis
                     subsp. mobilis (strain ATCC 31821 / ZM4 / CP4). Accession#:
                     P0DJA2"
     CDS             3167..4177
                     /gene="adhA"
                     /label="Alcohol dehydrogenase 1"
                     /note="Alcohol dehydrogenase 1 from Zymomonas mobilis
                     subsp. mobilis (strain ATCC 31821 / ZM4 / CP4). Accession#:
                     P20368"
     CDS             4211..4993
                     /gene="pfyP"
                     /label="Blue-light photoreceptor"
                     /note="Blue-light photoreceptor from Bacillus subtilis
                     (strain 168). Accession#: O34627"
     protein_bind    complement(5001..5034)
                     /label="FRT (minimal)"
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     CDS             5153..5965
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     protein_bind    complement(6062..6095)
                     /label="FRT (minimal)"
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     regulatory      6108..6159
                     /label="P22 terminator"
                     /note="P22 terminator"
                     /regulatory_class="terminator"
     promoter        6232..6260
                     /label="Pc promoter"
                     /note="class 1 integron promoter"
     CDS             6449..6979
                     /label="GmR"
                     /note="gentamycin acetyltransferase"
     rep_origin      8265..9034
                     /label="pBBR1 oriV"
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1
                     Rep protein for replication"
     CDS             9035..9694
                     /label="pBBR1 Rep"
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
     primer_bind     10404..10420
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        10430..10448
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gagctctact acgtcagtgt cgtgccatag atctttcaga atagcggaac acatagccgg
       61 tgctatactt aatctcgtta attactggga cataacggtc tctccacgca tccatcaaaa
      121 aagatgtgag cttgatcaaa aacaaaaaat atttcactcg acaggagtat ttatattgcg
      181 cggtcacctt aaaaaatcca cttaagaagg taggtgttac atgtcctata ctgtcggtac
      241 ctatttagcg gagcgtctgg tccagattgg tctgaagcat cacttcgcag tcgcgggcga
      301 ctacaacctc gtcctgctgg acaacctgct gctgaacaag aacatggagc aagtgtactg
      361 ctgtaacgaa ctcaattgcg gttttagcgc agagggttac gcacgcgcga aaggcgcggc
      421 ggctgcggtt gtcacttatt ctgtcggcgc cctgagcgct tttgatgcta tcggtggcgc
      481 ctatgcagaa aaccttccgg ttatcctgat ctccggtgct ccgaacaata acgatcatgc
      541 cgcaggtcac gtcctgcatc acgcgctggg caagacggac taccactacc agttggagat
      601 ggctaagaat atcactgcgg cggcggaggc tatctacacc ccggaggagg caccggcgaa
      661 gattgaccac gtgatcaaga ccgcactgcg tgagaagaag ccggtttatc tcgaaatcgc
      721 ttgcaacatt gcttccatgc cgtgcgccgc accaggtccg gcaagcgcgt tattcaacga
      781 cgaggcgagc gatgaagcaa gcctgaacgc tgctgtggaa gaaacgctca agtttatcgc
      841 taaccgcgat aaggttgcag tgttggtcgg ttcgaaactg cgtgccgcag gcgcagagga
      901 agccgccgta aaattcgcgg atgcattggg tggcgctgtt gcaacgatgg cggcagcgaa
      961 gagctttttc ccggaagaaa acccacatta catcggcacg tcgtggggtg aagtcagcta
     1021 cccgggtgtt gagaagacga tgaaggaggc cgacgcggtt attgccctgg ctccagtctt
     1081 caatgattac tcgacgaccg gctggaccga tattccggac ccaaaaaagc tggtcctggc
     1141 agaaccgcgt agcgttgtgg tgaatggtgt gcgttttcct agcgtccatc tgaaagatta
     1201 cctgacccgc ctggcccaaa aagtcagcaa gaaaaccggt gcgctggact ttttcaaatc
     1261 actgaacgcg ggtgaattga agaaagcggc ccctgcggat ccgagcgcac cgctggttaa
     1321 cgctgaaatt gctcgccagg tggaagcatt gctgaccccg aatacgaccg tgattgctga
     1381 aaccggtgac tcttggttca atgctcagcg catgaagctc ccgaacggtg ctcgcgtgga
     1441 atacgagatg cagtggggcc atatcggctg gtctgtcccg gcagcgttcg gttatgccgt
     1501 cggtgccccg gaacgtcgca acatcctcat ggttggtgat ggttccttcc agttgaccgc
     1561 gcaggaagtg gctcagatgg tgcgcctgaa actgccggtg atcatctttc tgattaacaa
     1621 ttatggttat accattgaag tgatgatcca cgacggtcct tataacaaca tcaaaaactg
     1681 ggattacgcg ggtttgatgg aagtgttcaa tggcaacggt ggttatgaca gcggcgcagg
     1741 taagggtctg aaagctaaga ccggtggtga gctggctgag gcgatcaagg tggccctggc
     1801 gaacactgat ggcccaacgt tgatcgagtg cttcatcggt cgtgaagact gcactgaaga
     1861 attggtcaaa tggggtaagc gcgttgctgc cgcgaattct cgtaagccgg tcaataagtt
     1921 gctgtaattt ttggggatca attccagctc ggtacccaat ctagtatgta gggtgaggtt
     1981 atagctatgg cctcgagcac gttctatatt ccgttcgtga acgagatggg cgaaggcagc
     2041 ttggagaagg ctattaaaga cctgaacggt agcggtttca aaaacgcact gatcgttagc
     2101 gatgccttta tgaacaagag cggcgtggtc aagcaggtgg cagacctgct gaaggcgcaa
     2161 ggtatcaatt cggccgttta tgacggtgtt atgccgaacc cgaccgttac ggcagttctg
     2221 gaaggcctga agatcctgaa ggataacaat agcgacttcg tcatctccct cggcggtggt
     2281 agcccacacg attgcgcaaa ggcgatcgcg ctggtcgcga cgaacggtgg cgaggtgaag
     2341 gactatgaag gtatcgacaa atctaagaaa cctgccctgc ctttgatgag catcaacacg
     2401 acggctggta ccgctagcga aatgacccgt ttctgcatca tcactgatga agtccgtcac
     2461 gttaagatgg ccattgttga ccgtcacgtt accccgatgg tgtcggtaaa tgatccgctg
     2521 ttgatggtgg gtatgcctaa gggtctgacc gccgctacgg gtatggatgc gctgactcac
     2581 gcatttgagg cgtacagctc cacggcggca acccctatca ccgacgcctg tgcactgaaa
     2641 gccgccagca tgatcgcgaa gaacctgaaa actgcctgcg acaatggtaa agatatgccg
     2701 gcacgtgagg cgatggcgta tgcgcagttc ctggctggta tggcgttcaa taacgcgagc
     2761 ctgggttatg tgcatgcgat ggcccatcag ctgggtggtt actataatct gccgcacggc
     2821 gtgtgtaacg ctgtcctgct gccgcatgtg ttagcctata atgcttccgt ggtcgcaggc
     2881 cgcctgaagg atgtgggtgt tgcgatgggt ctggatattg ccaatctggg tgacaaggag
     2941 ggcgccgagg caacgatcca ggccgtacgt gacttggcag catccattgg catcccggct
     3001 aacctgacgg aactgggcgc gaagaaagag gatgtgccgc ttcttgctga ccacgctctg
     3061 aaagatgctt gtgctctgac caacccgcgt cagggtgatc agaaggaggt cgaggagctg
     3121 ttcctgagcg cattctaacg ccatatatca acaaaaggta gtcaccatga aagctgcagt
     3181 tatcactaaa gatcatacga tcgaagtgaa agacaccaaa ttacgcccgc tgaaatatgg
     3241 cgaggctctg ctggagatgg aatattgcgg tgtctgtcac acggatctgc atgtcaaaaa
     3301 cggtgacttt ggtgatgaaa cgggtcgcat caccggccac gagggtattg gtatcgtgaa
     3361 acaggttggt gagggcgtta cttctctgaa agtcggtgat cgtgccagtg ttgcatggtt
     3421 cttcaaaggc tgcggccatt gtgaatactg tgttagcggt aacgaaaccc tgtgccgtaa
     3481 tgttgagaac gcgggttata cggtggatgg cgcgatggcg gaagagtgca ttgtggtcgc
     3541 ggactatagc gtcaaggtcc cggatggcct ggacccagcc gttgcgagca gcatcacctg
     3601 cgcgggtgta accacctata aagcagtcaa agtttctcag atacagccgg gacaatggct
     3661 ggctatctat ggcttgggcg gtttaggcaa tctggccctg caatatgcca agaatgtttt
     3721 caacgccaaa gtcatcgcga ttgatgtcaa cgacgaacaa ctggccttcg ccaaggaatt
     3781 gggtgcggat atggttatca acccgaaaaa cgaggatgcc gcgaagatca tccaagagaa
     3841 agtgggtggt gcgcacgcaa ccgtggtgac ggcagttgcc aaaagcgcgt tcaatagcgc
     3901 agtggaggct atccgcgcgg gtggtcgtgt cgtggcagta ggcctgccgc cggagaagat
     3961 ggacttatcg attccgcgcc tggtgctgga tggcatcgaa gttttgggta gcctggttgg
     4021 tacgcgtgag gatctgaaag aggcgttcca attcgcagct gagggtaagg tgaaaccgaa
     4081 agtgacgaag cgtaaggtcg aagaaatcaa ccaaattttt gacgagatgg agcacggcaa
     4141 gttcacgggc cgtatggttg tcgacttcac gcatcattaa cacttttaga gaaagaaggg
     4201 acagtcaagt atggcgtcct ttcagtcttt tggtattccg ggtcagctgg aggtgattaa
     4261 gaaggcgctg gatcatgtgc gcgttggcgt cgttatcacc gatccggccc tggaagataa
     4321 cccgattgtg tatgtgaatc agggcttcgt ccaaatgacc ggctacgaaa cggaagaaat
     4381 cttaggcaaa aatgcacgct tcctgcaagg caaacacacc gacccggcag aggtcgataa
     4441 cattcgcacc gcgctgcaaa ataaagaacc ggttacggtc cagatccaga actataaaaa
     4501 agacggcacg atgttctgga atgaactgaa cattgacccg atggagattg aggataagac
     4561 gtactttgtt ggtattcaga acgacatcac caaacagaag gagtacgaga agctgttgga
     4621 ggattctctg accgaaatca ctgcgctgag cactcctatc gttccgattc gtaatggtat
     4681 cagcgcgttg ccgctggttg gcaatctgac ggaggagaga tttaatagca tcgtgtgtac
     4741 tctgactaac atcctgagca ccagcaagga cgattatctg atcattgatc tgagcggcct
     4801 ggcgcaggtg aacgaacaga ccgcggacca aatcttcaaa ttgagccatc tgctgaaact
     4861 caccggtacg gaactgatca ttacgggtat caagccggaa ttggcgatga aaatgaataa
     4921 gttggacgca aacttctcct ctttgaaaac ctactctaat gttaaagacg ccgttaaagt
     4981 gctgccgatt atgtaagctt gaagttccta tactttctag agaataggaa cttcttttca
     5041 acaaagccac gttgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     5101 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgtctca
     5161 tattcaacgt gaaacctctt gttctcgtcc tcgtctgaat agcaacatgg acgctgatct
     5221 gtacggttac aagtgggcgc gtgataacgt gggccaatcc ggtgctacta tctaccgtct
     5281 gtatggtaaa ccggacgccc cggaactgtt cctgaaacac ggcaaaggct ctgtggcaaa
     5341 cgatgtaacc gacgaaatgg tgcgtctgaa ctggctgacc gagtttatgc cgctgccaac
     5401 catcaaacac tttatccgta ccccggacga tgcctggctg ctgactaccg ctattccggg
     5461 caaaaccgcc ttccaagttc tggaagaata cccagattcc ggcgaaaata tcgtagatgc
     5521 tctggctgta ttcctgcgtc gcctgcactc tattccggtg tgcaactgtc cgttcaactc
     5581 tgaccgcgta ttccgtctgg cccaagccca gtctcgtatg aataacggcc tggttgacgc
     5641 gagcgacttc gacgacgaac gcaacggttg gccggtggaa caggtttgga aagaaatgca
     5701 caaactgctg ccattctccc cagacagcgt tgtcacgcac ggtgatttta gcctggataa
     5761 cctgatcttt gatgaaggca aactgatcgg ttgtattgat gtaggtcgtg tgggcatcgc
     5821 agatcgttac caagacctgg ctatcctgtg gaactgcctg ggcgagttca gcccaagcct
     5881 gcaaaaacgc ctgttccaga agtacggtat cgacaacccg gacatgaata aactgcaatt
     5941 ccacctgatg ctggacgagt tcttttaaga attaattcat gagcggatac atatttgaat
     6001 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     6061 agaagttcct atactttcta gagaatagga acttcataaa tgcactcgtt aaataaagcc
     6121 ctgagtttaa ccgctcgggg ctttttgcgt tttaagcacg acatttctgg aagcacttag
     6181 caccacagcg gtacccgcac accgtggaaa cggatgaagg cacgaaccca gttgacataa
     6241 gcctgttcgg ttcgtaaact gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga
     6301 accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc ttgttatgac
     6361 tgtttttttg tacagtctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt
     6421 cgatgtttga tgttatggag cagcaacgat gttacgcagc agcaacgatg ttacgcagca
     6481 gggcagtcgc cctaaaacaa agttaggtgg ctcaagtatg ggcatcattc gcacatgtag
     6541 gctcggccct gaccaagtca aatccatgcg ggctgctctt gatcttttcg gtcgtgagtt
     6601 cggagacgta gccacctact cccaacatca gccggactcc gattacctcg ggaacttgct
     6661 ccgtagtaag acattcatcg cgcttgctgc cttcgaccaa gaagcggttg ttggcgctct
     6721 cgcggcttac gttctgccca ggtttgagca gccgcgtagt gagatctata tctatgatct
     6781 cgcagtctcc ggcgagcacc ggaggcaggg cattgccacc gcgctcatca atctcctcaa
     6841 gcatgaggcc aacgcgcttg gtgcttatgt gatctacgtg caagcagatt acggtgacga
     6901 tcccgcagtg gctctctata caaagttggg catacgggaa gaagtgatgc actttgatat
     6961 cgacccaagt accgccacct aacaattcgt tcaagccgag atcggcttcc cggccgcgga
     7021 gttgttcggt aaattgtcac aacgccgcca ggtggcactt ttcggggaaa tgtgcgcgcc
     7081 cgcgttcctg ctggcgctgg gcctgtttct ggcgctggac ttcccgctgt tccgtcagca
     7141 gcttttcgcc cacggccttg atgatcgcgg cggccttggc ctgcatatcc cgattcaacg
     7201 gccccagggc gtccagaacg ggcttcaggc gctcccgaag gtctcgggcc gtctcttggg
     7261 cttgatcggc cttcttgcgc atctcacgcg ctcctgcggc ggcctgtagg gcaggctcat
     7321 acccctgccg aaccgctttt gtcagccggt cggccacggc ttccggcgtc tcaacgcgct
     7381 ttgagattcc cagcttttcg gccaatccct gcggtgcata ggcgcgtggc tcgaccgctt
     7441 gcgggctgat ggtgacgtgg cccactggtg gccgctccag ggcctcgtag aacgcctgaa
     7501 tgcgcgtgtg acgtgccttg ctgccctcga tgccccgttg cagccctaga tcggccacag
     7561 cggccgcaaa cgtggtctgg tcgcgggtca tctgcgcttt gttgccgatg aactccttgg
     7621 ccgacagcct gccgtcctgc gtcagcggca ccacgaacgc ggtcatgtgc gggctggttt
     7681 cgtcacggtg gatgctggcc gtcacgatgc gatccgcccc gtacttgtcc gccagccact
     7741 tgtgcgcctt ctcgaagaac gccgcctgct gttcttggct ggccgacttc caccattccg
     7801 ggctggccgt catgacgtac tcgaccgcca acacagcgtc cttgcgccgc ttctctggca
     7861 gcaactcgcg cagtcggccc atcgcttcat cggtgctgct ggccgcccag tgctcgttct
     7921 ctggcgtcct gctggcgtca gcgttgggcg tctcgcgctc gcggtaggcg tgcttgagac
     7981 tggccgccac gttgcccatt ttcgccagct tcttgcatcg catgatcgcg tatgccgcca
     8041 tgcctgcccc tcccttttgg tgtccaaccg gctcgacggg ggcagcgcaa ggcggtgcct
     8101 ccggcgggcc actcaatgct tgagtatact cactagactt tgcttcgcaa agtcgtgacc
     8161 gcctacggcg gctgcggcgc cctacgggct tgctctccgg gcttcgccct gcgcggtcgc
     8221 tgcgctccct tgccagcccg tggatatgtg gacgatggcc gcgagcggcc accggctggc
     8281 tcgcttcgct cggcccgtgg acaaccctgc tggacaagct gatggacagg ctgcgcctgc
     8341 ccacgagctt gaccacaggg attgcccacc ggctacccag ccttcgacca catacccacc
     8401 ggctccaact gcgcggcctg cggccttgcc ccatcaattt ttttaatttt ctctggggaa
     8461 aagcctccgg cctgcggcct gcgcgcttcg cttgccggtt ggacaccaag tggaaggcgg
     8521 gtcaaggctc gcgcagcgac cgcgcagcgg cttggccttg acgcgcctgg aacgacccaa
     8581 gcctatgcga gtgggggcag tcgaaggcga agcccgcccg cctgcccccc gagcctcacg
     8641 gcggcgagtg cgggggttcc aagggggcag cgccaccttg ggcaaggccg aaggccgcgc
     8701 agtcgatcaa caagccccgg aggggccact ttttgccgga gggggagccg cgccgaaggc
     8761 gtgggggaac cccgcagggg tgcccttctt tgggcaccaa agaactagat atagggcgaa
     8821 atgcgaaaga cttaaaaatc aacaacttaa aaaagggggg tacgcaacag ctcattgcgg
     8881 caccccccgc aatagctcat tgcgtaggtt aaagaaaatc tgtaattgac tgccactttt
     8941 acgcaacgca taattgttgt cgcgctgccg aaaagttgca gctgattgcg catggtgccg
     9001 caaccgtgcg gcaccctacc gcatggagat aagcatggcc acgcagtcca gagaaatcgg
     9061 cattcaagcc aagaacaagc ccggtcactg ggtgcaaacg gaacgcaaag cgcatgaggc
     9121 gtgggccggg cttattgcga ggaaacccac ggcggcaatg ctgctgcatc acctcgtggc
     9181 gcagatgggc caccagaacg ccgtggtggt cagccagaag acactttcca agctcatcgg
     9241 acgttctttg cggacggtcc aatacgcagt caaggacttg gtggccgagc gctggatctc
     9301 cgtcgtgaag ctcaacggcc ccggcaccgt gtcggcctac gtggtcaatg accgcgtggc
     9361 gtggggccag ccccgcgacc agttgcgcct gtcggtgttc agtgccgccg tggtggttga
     9421 tcacgacgac caggacgaat cgctgttggg gcatggcgac ctgcgccgca tcccgaccct
     9481 gtatccgggc gagcagcaac taccgaccgg ccccggcgag gagccgccca gccagcccgg
     9541 cattccgggc atggaaccag acctgccagc cttgaccgaa acggaggaat gggaacggcg
     9601 cgggcagcag cgcctgccga tgcccgatga gccgtgtttt ctggacgatg gcgagccgtt
     9661 ggagccgccg acacgggtca cgctgccgcg ccggtagcac ttgggttgcg cagcaacccg
     9721 taagtgcgct gttccagact atcggctgta gccgcctcgc cgccctatac cttgtctgcc
     9781 tccccgcgtt gcgtcgcggt gcatggagcc gggccacctc gacctgaatg gaagccggcg
     9841 gcacctcgct aacggattca ccgtttttat caggctctgg gaggcagaat aaatgatcat
     9901 atcgtcaatt attacctcca cggggagagc ctgagcaaac tggcctcagg catttgagaa
     9961 gcacacggtc acactgcttc cggtagtcaa taaaccggta aaccagcaat agacataagc
    10021 ggctatttaa cgaccctgcc ctgaaccgac gaccgggtcg aatttgcttt cgaatttctg
    10081 ccattcatcc gcttattatc acttattcag gcgtagcacc aggcgtttaa gggcaccaat
    10141 aactgcctta aaaaaattac gccccgccct gccactcatc gcagtcggcc tattggttaa
    10201 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
    10261 tttccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
    10321 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
    10381 agggttttcc cagtcacgac gttgtaaaac gacggccagt gagcgcgcgt aatacgactc
    10441 actatagggc gaattg
//
