LOCUS       V004170                 7940 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004170
VERSION     V004170
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7940)
  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 7940)
  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 7940)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7940)
  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..7940
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    233..254
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        269..299
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    307..323
                     /label="lac operator"
                     /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)."
     primer_bind     331..347
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        368..386
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     416..432
                     /label="KS primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             558..2261
                     /gene="pdc"
                     /label="Pyruvate decarboxylase"
                     /note="Pyruvate decarboxylase from Zymomonas mobilis subsp.
                     mobilis (strain ATCC 31821 / ZM4 / CP4). Accession#:
                     P06672"
     CDS             2324..3472
                     /gene="adhB"
                     /label="Alcohol dehydrogenase 2"
                     /note="Alcohol dehydrogenase 2 from Zymomonas mobilis
                     subsp. mobilis (strain ATCC 31821 / ZM4 / CP4). Accession#:
                     P0DJA2"
     promoter        complement(3687..3705)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3715..3731)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             complement(4441..5100)
                     /label="pBBR1 Rep"
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
     rep_origin      complement(5101..5870)
                     /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"
     CDS             complement(7156..7686)
                     /label="GmR"
                     /note="gentamycin acetyltransferase"
     promoter        complement(7875..7903)
                     /label="Pc promoter"
                     /note="class 1 integron promoter"
ORIGIN
        1 ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc
       61 atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtttt
      121 tatgcatgcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
      181 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
      241 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
      301 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
      361 agcgcgcaat taaccctcac taaagggaac aaaagctggg taccgggccc cccctcgagg
      421 tcgacggtat cgataagctt gatatcgaat tcgagctcgg tacccaaaaa tgcctatagc
      481 taaatccgga acgacacttt agaggtttct gggtcatcct gattcagaca tagtgttttg
      541 aatatatgga gtaagcaatg agttatactg tcggtaccta tttagcggag cggcttgtcc
      601 agattggtct caagcatcac ttcgcagtcg cgggcgacta caacctcgtc cttcttgaca
      661 acctgctttt gaacaaaaac atggagcagg tttattgctg taacgaactg aactgcggtt
      721 tcagtgcaga aggttatgct cgtgccaaag gcgcagcagc agccgtcgtt acctacagcg
      781 tcggtgcgct ttccgcattt gatgctatcg gtggcgccta tgcagaaaac cttccggtta
      841 tcctgatctc cggtgctccg aacaacaatg atcacgctgc tggtcacgtg ttgcatcacg
      901 ctcttggcaa aaccgactat cactatcagt tggaaatggc caagaacatc acggccgccg
      961 ctgaagcgat ttacaccccg gaagaagctc cggctaaaat cgatcacgtg attaaaactg
     1021 ctcttcgtga gaagaagccg gtttatctcg aaatcgcttg caacattgct tccatgccct
     1081 gcgccgctcc tggaccggca agcgcattgt tcaatgacga agccagcgac gaagcttctt
     1141 tgaatgcagc ggttgaagaa accctgaaat tcatcgccaa ccgcgacaaa gttgccgtcc
     1201 tcgtcggcag caagctgcgc gcagctggtg ctgaagaagc tgctgtcaaa tttgctgatg
     1261 ctctcggtgg cgcagttgct accatggctg ctgcaaaaag cttcttccca gaagaaaacc
     1321 cgcattacat cggcacctca tggggtgaag tcagctatcc gggcgttgaa aagacgatga
     1381 aagaagccga tgcggttatc gctctggctc ctgtcttcaa cgactactcc accactggtt
     1441 ggacggatat tcctgatcct aagaaactgg ttctcgctga accgcgttct gtcgtcgtta
     1501 acggcattcg cttccccagc gtccatctga aagactatct gacccgtttg gctcagaaag
     1561 tttccaagaa aaccggtgca ttggacttct tcaaatccct caatgcaggt gaactgaaga
     1621 aagccgctcc ggctgatccg agtgctccgt tggtcaacgc agaaatcgcc cgtcaggtcg
     1681 aagctcttct gaccccgaac acgacggtta ttgctgaaac cggtgactct tggttcaatg
     1741 ctcagcgcat gaagctcccg aacggtgctc gcgttgaata tgaaatgcag tggggtcaca
     1801 ttggttggtc cgttcctgcc gccttcggtt atgccgtcgg tgctccggaa cgtcgcaaca
     1861 tcctcatggt tggtgatggt tccttccagc tgacggctca ggaagtcgct cagatggttc
     1921 gcctgaaact gccggttatc atcttcttga tcaataacta tggttacacc atcgaagtta
     1981 tgatccatga tggtccgtac aacaacatca agaactggga ttatgccggt ctgatggaag
     2041 tgttcaacgg taacggtggt tatgacagcg gtgctggtaa aggcctgaag gctaaaaccg
     2101 gtggcgaact ggcagaagct atcaaggttg ctctggcaaa caccgacggc ccaaccctga
     2161 tcgaatgctt catcggtcgt gaagactgca ctgaagaatt ggtcaaatgg ggtaagcgcg
     2221 ttgctgccgc caacagccgt aagcctgtta acaagctcct ctagtttttg gggatcaatt
     2281 cgagctcggt acccaaacta gtatgtaggg tgaggttata gctatggctt cttcaacttt
     2341 ttatattcct ttcgtcaacg aaatgggcga aggttcgctt gaaaaagcaa tcaaggatct
     2401 taacggcagc ggctttaaaa atgcgctgat cgtttctgat gctttcatga acaaatccgg
     2461 tgttgtgaag caggttgctg acctgttgaa agcacagggt attaattctg ctgtttatga
     2521 tggcgttatg ccgaacccga ctgttaccgc agttctggaa ggccttaaga tcctgaagga
     2581 taacaattca gacttcgtca tctccctcgg tggtggttct ccccatgact gcgccaaagc
     2641 catcgctctg gtcgcaacca atggtggtga agtcaaagac tacgaaggta tcgacaaatc
     2701 taagaaacct gccctgcctt tgatgtcaat caacacgacg gctggtacgg cttctgaaat
     2761 gacgcgtttc tgcatcatca ctgatgaagt ccgtcacgtt aagatggcca ttgttgaccg
     2821 tcacgttacc ccgatggttt ccgtcaacga tcctctgttg atggttggta tgccaaaagg
     2881 cctgaccgcc gccaccggta tggatgctct gacccacgca tttgaagctt attcttcaac
     2941 ggcagctact ccgatcaccg atgcttgcgc tttgaaagca gcttccatga tcgctaagaa
     3001 tctgaagacc gcttgcgaca acggtaagga tatgccggct cgtgaagcta tggcttatgc
     3061 ccaattcctc gctggtatgg ccttcaacaa cgcttcgctt ggttatgtcc atgctatggc
     3121 tcaccagttg ggcggttact acaacctgcc gcatggtgtc tgcaacgctg ttctgcttcc
     3181 gcatgttctg gcttataacg cctctgtcgt tgctggtcgt ctgaaagacg ttggtgttgc
     3241 tatgggtctc gatatcgcca atctcggtga taaagaaggc gcagaagcca ccattcaggc
     3301 tgttcgcgat ctggctgctt ccattggtat tccagcaaac ctgaccgagc tgggtgctaa
     3361 gaaagaagat gtgccgcttc ttgctgacca cgctctgaaa gatgcttgtg ctctgaccaa
     3421 cccgcgtcag ggtgatcaga aagaagttga agaactcttc ctgagcgctt tctaatttca
     3481 aaacaggaaa acggttttcc gtcctgtctt gattttcaag caaacaatgc ctccgatttc
     3541 taatcggagg catttgtttt tgtttattgc aaaaacaaaa aatattgtta caaattttta
     3601 caggctatta agcctaccgt cataaataat ttgccatttg gggatccctc tagagcggcc
     3661 gccaccgcgg tggagctcca attcgcccta tagtgagtcg tattacgcgc gctcactggc
     3721 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     3781 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     3841 ccaacagttg cgcagcctga atggcgaatg gaaattgtaa gcgttaatat tttgttaaaa
     3901 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga ctgcgatgag
     3961 tggcagggcg gggcgtaatt tttttaaggc agttattggt gcccttaaac gcctggtgct
     4021 acgcctgaat aagtgataat aagcggatga atggcagaaa ttcgaaagca aattcgaccc
     4081 ggtcgtcggt tcagggcagg gtcgttaaat agccgcttat gtctattgct ggtttaccgg
     4141 tttattgact accggaagca gtgtgaccgt gtgcttctca aatgcctgag gccagtttgc
     4201 tcaggctctc cccgtggagg taataattga cgatatgatc atttattctg cctcccagag
     4261 cctgataaaa acggtgaatc cgttagcgag gtgccgccgg cttccattca ggtcgaggtg
     4321 gcccggctcc atgcaccgcg acgcaacgcg gggaggcaga caaggtatag ggcggcgagg
     4381 cggctacagc cgatagtctg gaacagcgca cttacgggtt gctgcgcaac ccaagtgcta
     4441 ccggcgcggc agcgtgaccc gtgtcggcgg ctccaacggc tcgccatcgt ccagaaaaca
     4501 cggctcatcg ggcatcggca ggcgctgctg cccgcgccgt tcccattcct ccgtttcggt
     4561 caaggctggc aggtctggtt ccatgcccgg aatgccgggc tggctgggcg gctcctcgcc
     4621 ggggccggtc ggtagttgct gctcgcccgg atacagggtc gggatgcggc gcaggtcgcc
     4681 atgccccaac agcgattcgt cctggtcgtc gtgatcaacc accacggcgg cactgaacac
     4741 cgacaggcgc aactggtcgc ggggctggcc ccacgccacg cggtcattga ccacgtaggc
     4801 cgacacggtg ccggggccgt tgagcttcac gacggagatc cagcgctcgg ccaccaagtc
     4861 cttgactgcg tattggaccg tccgcaaaga acgtccgatg agcttggaaa gtgtcttctg
     4921 gctgaccacc acggcgttct ggtggcccat ctgcgccacg aggtgatgca gcagcattgc
     4981 cgccgtgggt ttcctcgcaa taagcccggc ccacgcctca tgcgctttgc gttccgtttg
     5041 cacccagtga ccgggcttgt tcttggcttg aatgccgatt tctctggact gcgtggccat
     5101 gcttatctcc atgcggtagg gtgccgcacg gttgcggcac catgcgcaat cagctgcaac
     5161 ttttcggcag cgcgacaaca attatgcgtt gcgtaaaagt ggcagtcaat tacagatttt
     5221 ctttaaccta cgcaatgagc tattgcgggg ggtgccgcaa tgagctgttg cgtacccccc
     5281 ttttttaagt tgttgatttt taagtctttc gcatttcgcc ctatatctag ttctttggtg
     5341 cccaaagaag ggcacccctg cggggttccc ccacgccttc ggcgcggctc cccctccggc
     5401 aaaaagtggc ccctccgggg cttgttgatc gactgcgcgg ccttcggcct tgcccaaggt
     5461 ggcgctgccc ccttggaacc cccgcactcg ccgccgtgag gctcgggggg caggcgggcg
     5521 ggcttcgcct tcgactgccc ccactcgcat aggcttgggt cgttccaggc gcgtcaaggc
     5581 caagccgctg cgcggtcgct gcgcgagcct tgacccgcct tccacttggt gtccaaccgg
     5641 caagcgaagc gcgcaggccg caggccggag gcttttcccc agagaaaatt aaaaaaattg
     5701 atggggcaag gccgcaggcc gcgcagttgg agccggtggg tatgtggtcg aaggctgggt
     5761 agccggtggg caatccctgt ggtcaagctc gtgggcaggc gcagcctgtc catcagcttg
     5821 tccagcaggg ttgtccacgg gccgagcgaa gcgagccagc cggtggccgc tcgcggccat
     5881 cgtccacata tccacgggct ggcaagggag cgcagcgacc gcgcagggcg aagcccggag
     5941 agcaagcccg tagggcgccg cagccgccgt aggcggtcac gactttgcga agcaaagtct
     6001 agtgagtata ctcaagcatt gagtggcccg ccggaggcac cgccttgcgc tgcccccgtc
     6061 gagccggttg gacaccaaaa gggaggggca ggcatggcgg catacgcgat catgcgatgc
     6121 aagaagctgg cgaaaatggg caacgtggcg gccagtctca agcacgccta ccgcgagcgc
     6181 gagacgccca acgctgacgc cagcaggacg ccagagaacg agcactgggc ggccagcagc
     6241 accgatgaag cgatgggccg actgcgcgag ttgctgccag agaagcggcg caaggacgct
     6301 gtgttggcgg tcgagtacgt catgacggcc agcccggaat ggtggaagtc ggccagccaa
     6361 gaacagcagg cggcgttctt cgagaaggcg cacaagtggc tggcggacaa gtacggggcg
     6421 gatcgcatcg tgacggccag catccaccgt gacgaaacca gcccgcacat gaccgcgttc
     6481 gtggtgccgc tgacgcagga cggcaggctg tcggccaagg agttcatcgg caacaaagcg
     6541 cagatgaccc gcgaccagac cacgtttgcg gccgctgtgg ccgatctagg gctgcaacgg
     6601 ggcatcgagg gcagcaaggc acgtcacacg cgcattcagg cgttctacga ggccctggag
     6661 cggccaccag tgggccacgt caccatcagc ccgcaagcgg tcgagccacg cgcctatgca
     6721 ccgcagggat tggccgaaaa gctgggaatc tcaaagcgcg ttgagacgcc ggaagccgtg
     6781 gccgaccggc tgacaaaagc ggttcggcag gggtatgagc ctgccctaca ggccgccgca
     6841 ggagcgcgtg agatgcgcaa gaaggccgat caagcccaag agacggcccg agaccttcgg
     6901 gagcgcctga agcccgttct ggacgccctg gggccgttga atcgggatat gcaggccaag
     6961 gccgccgcga tcatcaaggc cgtgggcgaa aagctgctga cggaacagcg ggaagtccag
     7021 cgccagaaac aggcccagcg ccagcaggaa cgcgggcgcg cacatttccc cgaaaagtgc
     7081 cacctggcgg cgttgtgaca atttaccgaa caactccgcg gccgggaagc cgatctcggc
     7141 ttgaacgaat tgttaggtgg cggtacttgg gtcgatatca aagtgcatca cttcttcccg
     7201 tatgcccaac tttgtataga gagccactgc gggatcgtca ccgtaatctg cttgcacgta
     7261 gatcacataa gcaccaagcg cgttggcctc atgcttgagg agattgatga gcgcggtggc
     7321 aatgccctgc ctccggtgct cgccggagac tgcgagatca tagatataga tctcactacg
     7381 cggctgctca aacctgggca gaacgtaagc cgcgagagcg ccaacaaccg cttcttggtc
     7441 gaaggcagca agcgcgatga atgtcttact acggagcaag ttcccgaggt aatcggagtc
     7501 cggctgatgt tgggagtagg tggctacgtc tccgaactca cgaccgaaaa gatcaagagc
     7561 agcccgcatg gatttgactt ggtcagggcc gagcctacat gtgcgaatga tgcccatact
     7621 tgagccacct aactttgttt tagggcgact gccctgctgc gtaacatcgt tgctgctgcg
     7681 taacatcgtt gctgctccat aacatcaaac atcgacccac ggcgtaacgc gcttgctgct
     7741 tggatgcccg aggcatagac tgtacaaaaa aacagtcata acaagccatg aaaaccgcca
     7801 ctgcgccgtt accaccgctg cgttcggtca aggttctgga ccagttgcgt gagcgcatac
     7861 gctacttgca ttacagttta cgaaccgaac aggcttatgt caactgggtt cgtgccttca
     7921 tccgtttcca cggtgtgcgt
//
