LOCUS       V004174                 7284 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004174
VERSION     V004174
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7284)
  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 7284)
  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 7284)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7284)
  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..7284
                     /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."
     regulatory      4154..4182
                     /label="Promoter#25"
                     /note="Promoter#25"
                     /regulatory_class="promoter"
     regulatory      4236..4244
                     /label="pdc RBS#3"
                     /note="pdc RBS#3"
                     /regulatory_class="ribosome_binding_site"
     CDS             4250..5953
                     /gene="pdc"
                     /label="Pyruvate decarboxylase"
                     /note="Pyruvate decarboxylase from Zymomonas mobilis subsp.
                     mobilis (strain ATCC 31821 / ZM4 / CP4). Accession#:
                     P06672"
     regulatory      5979..5987
                     /label="adhB RBS#1"
                     /note="adhB RBS#1"
                     /regulatory_class="ribosome_binding_site"
     CDS             5993..7141
                     /gene="adhB"
                     /label="Alcohol dehydrogenase 2"
                     /note="Alcohol dehydrogenase 2 from Zymomonas mobilis
                     subsp. mobilis (strain ATCC 31821 / ZM4 / CP4). Accession#:
                     P0DJA2"
     regulatory      7188..7239
                     /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 ctctttacaa ttaatcatcc ggctcgcata atgtgtggaa ttggtagagt
     4201 atttttattg cgcggtcacc ttaaatccac tatcttaagg agatgttaca tgtcctatac
     4261 tgtcggtacc tatttagcgg agcgtctggt ccagattggt ctgaagcatc acttcgcagt
     4321 cgcgggcgac tacaacctcg tcctgctgga caacctgctg ctgaacaaga acatggagca
     4381 agtgtactgc tgtaacgaac tcaattgcgg ttttagcgca gagggttacg cacgcgcgaa
     4441 aggcgcggcg gctgcggttg tcacttattc tgtcggcgcc ctgagcgctt ttgatgctat
     4501 cggtggcgcc tatgcagaaa accttccggt tatcctgatc tccggtgctc cgaacaataa
     4561 cgatcatgcc gcaggtcacg tcctgcatca cgcgctgggc aagacggact accactacca
     4621 gttggagatg gctaagaata tcactgcggc ggcggaggct atctacaccc cggaggaggc
     4681 accggcgaag attgaccacg tgatcaagac cgcactgcgt gagaagaagc cggtttatct
     4741 cgaaatcgct tgcaacattg cttccatgcc gtgcgccgca ccaggtccgg caagcgcgtt
     4801 attcaacgac gaggcgagcg atgaagcaag cctgaacgct gctgtggaag aaacgctcaa
     4861 gtttatcgct aaccgcgata aggttgcagt gttggtcggt tcgaaactgc gtgccgcagg
     4921 cgcagaggaa gccgccgtaa aattcgcgga tgcattgggt ggcgctgttg caacgatggc
     4981 ggcagcgaag agctttttcc cggaagaaaa cccacattac atcggcacgt cgtggggtga
     5041 agtcagctac ccgggtgttg agaagacgat gaaggaggcc gacgcggtta ttgccctggc
     5101 tccagtcttc aatgattact cgacgaccgg ctggaccgat attccggacc caaaaaagct
     5161 ggtcctggca gaaccgcgta gcgttgtggt gaatggtgtg cgttttccta gcgtccatct
     5221 gaaagattac ctgacccgcc tggcccaaaa agtcagcaag aaaaccggtg cgctggactt
     5281 tttcaaatca ctgaacgcgg gtgaattgaa gaaagcggcc cctgcggatc cgagcgcacc
     5341 gctggttaac gctgaaattg ctcgccaggt ggaagcattg ctgaccccga atacgaccgt
     5401 gattgctgaa accggtgact cttggttcaa tgctcagcgc atgaagctcc cgaacggtgc
     5461 tcgcgtggaa tacgagatgc agtggggcca tatcggctgg tctgtcccgg cagcgttcgg
     5521 ttatgccgtc ggtgccccgg aacgtcgcaa catcctcatg gttggtgatg gttccttcca
     5581 gttgaccgcg caggaagtgg ctcagatggt gcgcctgaaa ctgccggtga tcatctttct
     5641 gattaacaat tatggttata ccattgaagt gatgatccac gacggtcctt ataacaacat
     5701 caaaaactgg gattacgcgg gtttgatgga agtgttcaat ggcaacggtg gttatgacag
     5761 cggcgcaggt aagggtctga aagctaagac cggtggtgag ctggctgagg cgatcaaggt
     5821 ggccctggcg aacactgatg gcccaacgtt gatcgagtgc ttcatcggtc gtgaagactg
     5881 cactgaagaa ttggtcaaat ggggtaagcg cgttgctgcc gcgaattctc gtaagccggt
     5941 caataagttg ctgtaatttt tgcaatctag tatggcctta ggggggatag ctatggcctc
     6001 gagcacgttc tatattccgt tcgtgaacga gatgggcgaa ggcagcttgg agaaggctat
     6061 taaagacctg aacggtagcg gtttcaaaaa cgcactgatc gttagcgatg cctttatgaa
     6121 caagagcggc gtggtcaagc aggtggcaga cctgctgaag gcgcaaggta tcaattcggc
     6181 cgtttatgac ggtgttatgc cgaacccgac cgttacggca gttctggaag gcctgaagat
     6241 cctgaaggat aacaatagcg acttcgtcat ctccctcggc ggtggtagcc cacacgattg
     6301 cgcaaaggcg atcgcgctgg tcgcgacgaa cggtggcgag gtgaaggact atgaaggtat
     6361 cgacaaatct aagaaacctg ccctgccttt gatgagcatc aacacgacgg ctggtaccgc
     6421 tagcgaaatg acccgtttct gcatcatcac tgatgaagtc cgtcacgtta agatggccat
     6481 tgttgaccgt cacgttaccc cgatggtgtc ggtaaatgat ccgctgttga tggtgggtat
     6541 gcctaagggt ctgaccgccg ctacgggtat ggatgcgctg actcacgcat ttgaggcgta
     6601 cagctccacg gcggcaaccc ctatcaccga cgcctgtgca ctgaaagccg ccagcatgat
     6661 cgcgaagaac ctgaaaactg cctgcgacaa tggtaaagat atgccggcac gtgaggcgat
     6721 ggcgtatgcg cagttcctgg ctggtatggc gttcaataac gcgagcctgg gttatgtgca
     6781 tgcgatggcc catcagctgg gtggttacta taatctgccg cacggcgtgt gtaacgctgt
     6841 cctgctgccg catgtgttag cctataatgc ttccgtggtc gcaggccgcc tgaaggatgt
     6901 gggtgttgcg atgggtctgg atattgccaa tctgggtgac aaggagggcg ccgaggcaac
     6961 gatccaggcc gtacgtgact tggcagcatc cattggcatc ccggctaacc tgacggaact
     7021 gggcgcgaag aaagaggatg tgccgcttct tgctgaccac gctctgaaag atgcttgtgc
     7081 tctgaccaac ccgcgtcagg gtgatcagaa ggaggtcgag gagctgttcc tgagcgcatt
     7141 ctaaatcgaa ttcctgcagc ccgggggatc cctagttcta gaaataagtt aaataaagcc
     7201 ctgagtttaa ccgctcgggg ctttttgcgt tttaagcaca ctagagcggc catgcatccc
     7261 agagcctgat aaaaacggtg aatc
//
