LOCUS       V009694                 8046 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V009694
VERSION     V009694
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8046)
  AUTHORS   Al-Hinai MA, Fast AG, Papoutsakis ET.
  TITLE     Novel system for efficient isolation of clostridium double-crossover
            allelic exchange mutants enabling markerless chromosomal gene
            deletions and DNA integration
  JOURNAL   Appl. Environ. Microbiol. 78 (22), 8112-8121 (2012)
   PUBMED   22983967
REFERENCE   2  (bases 1 to 8046)
  AUTHORS   Al-Hinai MA, Papoutsakis ET, Fast AG.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-SEP-2012) Biological Sciences, University of Delaware,
            15 Innovation Way, Newark, DE 19711, USA
REFERENCE   3  (bases 1 to 8046)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8046)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
            Environ. Microbiol."; date: "2012"; volume: "78"; issue: "22";
            pages: "8112-8121"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (06-SEP-2012) Biological Sciences, University of Delaware, 15
            Innovation Way, Newark, DE 19711, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8046
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     381..397
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             complement(465..905)
                     /codon_start=1
                     /gene="repL"
                     /product="gram positive replication protein"
                     /label="repL"
                     /note="RepL"
                     /protein_id="AFS68390.1"
                     /translation="MKERYGTVYKGSQRLIDEESGEVIEVDKLYRKQTSGNFVKAYIVQ
                     LISMLDMIGGKKLKIVNYILDNVHLSNNTMIATTREIAKATGTSLQTVITTLKILEEGN
                     IIKRKTGVLMLNPELLMRGDDQKQKYLLLEFGNFEQEANEID"
     gene            complement(465..905)
                     /gene="repL"
                     /label="repL"
     CDS             complement(1356..2087)
                     /gene="ermC'"
                     /label="rRNA adenine N-6-methyltransferase"
                     /note="rRNA adenine N-6-methyltransferase from Bacillus
                     subtilis. Accession#: P13956"
     CDS             complement(2667..3506)
                     /codon_start=1
                     /gene="bgaR"
                     /product="beta-galactosidase regulator"
                     /label="bgaR"
                     /note="BgaR"
                     /protein_id="AFS68392.1"
                     /translation="MQILWKKYVKENFEMNVDECGIEQGIPGLGYNYEVLKNAVIHYVT
                     KGYGTFKFNGKVYNLKQGDIFILLKGMQVEYVASIDDPWEYYWIGFSGSNANEYLNRTS
                     ITNSCVANCEENSKIPQIILNMCEISKTYNPSRSDDILLLKELYSLLYALIEEFPKPFE
                     YKDKELHTYIQDALNFINSNYMHSITVQEIADYVNLSRSYLYKMFIKNLGISPQRYLIN
                     LRMYKATLLLKSTKLPIGEVASSVGYSDSLLFSKTFSKHFSMSPLNYRNNQVNKPSI"
     gene            complement(2667..3506)
                     /gene="bgaR"
                     /label="bgaR"
     gene            3923..4022
                     /gene="repLAS"
                     /label="repLAS"
                     /note="antisense RNA to the repL"
     CDS             4360..5628
                     /label="FLP"
                     /note="site-specific recombinase"
     primer_bind     complement(5825..5841)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(5849..5865)
                     /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)."
     promoter        complement(5873..5903)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5918..5939)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6227..6815)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6989..7846)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7847..7951)
                     /label="AmpR promoter"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcg
      241 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
      301 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      361 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagcttgagc ttaatcgcat
      421 ttcatagatt gacctcccaa taactacgtg gtgttattgg gaggtcaatc tatttcattt
      481 gcctcttgct caaagttccc aaattcgagt aagaggtatt tttgtttttg gtcgtcgcct
      541 ctcattagta gttcagggtt taacattaat actccagttt ttctttttat aatatttcct
      601 tcttctaaga ttttaagtgt tgttattact gtttgtagac ttgttcctgt agcttttgct
      661 atttctcttg ttgtagctat cattgtattg ttacttaagt ggacattatc taggatatag
      721 ttaacgattt taagtttttt tccgccaatc atatctaaca tacttattaa ttgcactata
      781 tatgccttta cgaagttacc agacgtttgt ttacggtata acttgtctac ctctatgact
      841 tctccacttt cttcgtctat gagcctctga gagcctttat agactgttcc atatctttct
      901 ttcatctttt tctcactcct tattttaaac tattctaact atatcataac tgttctaaaa
      961 aaaaaagaac atttgttaaa agaaattaga acaaaatgag tgaaaaatta gaacaaacaa
     1021 attccttata aaccttatca tctcaaccta tattaagatt ttacctagtt gaatcttctt
     1081 ttctatataa agcgtcggag catatcaggg ggttatctaa cgtaaatgct acccttcggc
     1141 tcgctttcgc tcggcattga cgtcagatac tgcaccccct gaacccccat gctccaacag
     1201 caaaaaggaa actttttgct gcttttccga cgcttattcg cttcgctcat atttatatag
     1261 aaaagaagtg aatgcgcaaa agacataatc gattcacaaa aaataggtac acgaaaaaca
     1321 agttaaggga tgcagtttat gcatccctta acttacttat taaataattt atagctattg
     1381 aaaagagata agaattgttc aaagctaata ttgtttaaat cgtcaattcc tgcatgtttt
     1441 aaggaattgt taaattgatt ttttgtaaat attttcttgt attctttgtt aacccatttc
     1501 ataacgaaat aattatactt ctgtttatct ttgtgtgata ttcttgattt ttttctattt
     1561 aatctgataa gtgagctatt cactttaggt ttaggatgaa aatattctct tggaaccata
     1621 cttaatatag aaatatcaac ttctgccatt aaaaataatg ccaatgagcg ttttgtattt
     1681 aataatcttt tagcaaaccc gtattccacg attaaataaa tctcatcagc tatactatca
     1741 aaaacaattt tgcgtattat atccgtactt atgttataag gtatattacc aaatatttta
     1801 taggattggt ttttaggaaa tttaaactgc aatatatcct tgtttaaaac ttggaaatta
     1861 tcgtgatcaa caagtttatt ttctgtagtt ttgcataatt tatggtctat ttcaatggca
     1921 gttacgaaat tacacctctg tactaattca agggtaaaat gcccttttcc tgagccgatt
     1981 tcaaagatat tatcatgttc atttaatctt atatttgtca ttattttatc tatattatgt
     2041 tttgaagtaa taaagttttg actgtgtttt atatttttct cgttcattat aaccctcttt
     2101 attttttcct ccttataaaa ttagtataat tatagcacga gctctgataa atatgaacat
     2161 gatgagtgat cgttaaattt atactgcaat ctgatgcgat tattgaataa aagatatgag
     2221 agatttatct agtttctttt tttacaagaa aaaagaaagt tcttaaaggt tttatacttt
     2281 tggtcgtaga gcacacggtt taacgactta attacgaagt aaataagtct agtgtgttag
     2341 actttaatgt ttttttaagg cattagtgca tttaagcgtc agagcatggc tttatgccga
     2401 gaaaactatt ggttggaatg gcgtgtgtgt tagccaaagc tccccctcga gatgaaaagt
     2461 attagggcta atttagatat taattctaaa ttaagtgaaa ttaatatagt aattatatta
     2521 aaagtctaat taagacttta gaaaataaat ttcatataat tttaattctt ataaaagatt
     2581 taaaaagtat aacagaggtt ttaatttacg cctctgttat actttttatt tttgaaattt
     2641 ttttgtttta aagctgtatt ttaaatttat atacttggtt tatttacttg attatttctg
     2701 taatttagtg gagacattga aaaatgtttt gaaaaagttt ttgaaaataa cagggagtca
     2761 ctataaccta cactacttgc gacttctcct ataggaagtt tagtgctttt taataaaagg
     2821 gtggctttgt acattctaag gtttattaaa tatctttgag gagaaattcc aaggtttttt
     2881 atgaacattt tatataaata acttctactt aagttcacat aatcagcaat ttcttgaaca
     2941 gttatgctat gcatgtaatt agaattaatg aaattaagag catcttgaat atatgtgtgt
     3001 aattccttat ctttgtattc aaaaggtttt gggaattctt ctataagtgc gtacaataat
     3061 gagtaaagtt cttttagtaa tagtatgtca tcagatcttg aaggattata agtttttgat
     3121 atttcgcaca tatttaatat tatctgtgga atttttgagt tttcttcaca attagcaaca
     3181 caggagttag taatagaagt tctatttaaa tactcattag catttgaacc actaaatcct
     3241 atccagtagt attcccaagg atcatcaata gaagccacat actcaacttg catacctttt
     3301 agtagtataa aaatatcacc ttgttttaag ttatatacct taccattaaa tttaaaagtt
     3361 ccatatccct tagttacgta atgaataaca gcatttttca atacttcata gttatatcct
     3421 aatcctggta taccttgttc tataccacat tcatctacat tcatttcaaa gttttcttta
     3481 acatactttt tccacaatat ttgcatttct acctcctaac ctataaaatt agccaatttt
     3541 atagtagtct tatattaaac atttacatga gagctttgca aagcagttta tcaacataaa
     3601 agctttttat tttaaaataa attcttctaa atataagaat attttaaaga aatatcttta
     3661 tatattagtt attaaaattt ataagattat aagaaacatt ataacatatt ttagaacttt
     3721 ttaactattc taaaagatta atttacatat taacatttaa ttatgggtaa aaactatttt
     3781 gaaaaatgat ttatatggaa ttatgtttct taaatataca atcatgtttc atgaatacat
     3841 aattatttta aatgtattgg gagggtaaaa tgatattaaa aaatgaatac catgaagata
     3901 ctgcagaatc tagaatccgc ggtttgttta cggtataact tgtctacctc tatgacttct
     3961 ccactttctt cgtctatgag cctctgagag cctttataga ctgttccata tctttctttc
     4021 atggcgccac ttaatgattt gccagtaaaa gagattgttt ctagctctca cattcttgca
     4081 gatataatat tgcctagagc tgaagttata tatgattatc ttaagtaata aaaataagag
     4141 ttaccttaaa tggtaactct tattttttta atgtcgactc atagaattcg taatcatggt
     4201 cggtcgactg tggatggagt taagtcagca gaaagtataa tgagaaaata taaaatataa
     4261 ataattttct aaaaaactta acttcatgtg aaaagtttgt taaaatataa atgagcacgt
     4321 taatcattta acatagataa ttggatccaa ggaggttgta tgccacaatt tgatatatta
     4381 tgtaaaacac cacctaaggt gcttgttcgt cagtttgtgg aaaggtttga aagaccttca
     4441 ggtgagaaaa tagcattatg tgctgctgaa ctaacctatt tatgttggat gattacacat
     4501 aacggaacag caatcaagag agccacattc atgagctata atactatcat aagcaattcg
     4561 ctgagtttcg atattgtcaa taaatcactc cagtttaaat acaagacgca aaaagcaaca
     4621 attctggaag cctcattaaa gaaattgatt cctgcttggg aatttacaat tattccttac
     4681 tatggacaaa aacatcaatc tgatatcact gatattgtaa gtagtttgca attacagttc
     4741 gaatcatcgg aagaagcaga taagggaaat agccacagta aaaaaatgct taaagcactt
     4801 ctaagtgagg gtgaaagcat ctgggagatc actgagaaaa tactaaattc gtttgagtat
     4861 acttcgagat ttacaaaaac aaaaacttta taccaattcc tcttcctagc tactttcatc
     4921 aattgtggaa gattcagcga tattaagaac gttgatccga aatcatttaa attagtccaa
     4981 aataagtatc tgggagtaat aatccagtgt ttagtgacag agacaaagac aagcgttagt
     5041 aggcacatat acttctttag cgcaaggggt aggatcgatc cacttgtata tttggatgaa
     5101 tttttgagga attctgaacc agtcctaaaa cgagtaaata ggaccggcaa ttcttcaagc
     5161 aataaacagg aataccaatt attaaaagat aacttagtca gatcgtacaa taaagctttg
     5221 aagaaaaatg cgccttattc aatctttgct ataaaaaatg gcccaaaatc tcacattgga
     5281 agacatttga tgacctcatt tctttcaatg aagggcctaa cggagttgac taatgttgtg
     5341 ggaaattgga gcgataagcg tgcttctgcc gtggccagga caacgtatac tcatcagata
     5401 acagcaatac ctgatcacta cttcgcacta gtttctcggt actatgcata tgatccaata
     5461 tcaaaggaaa tgatagcatt gaaggatgag actaatccaa ttgaggagtg gcagcatata
     5521 gaacagctaa agggtagtgc tgaaggaagc atacgatacc ccgcatggaa tgggataata
     5581 tcacaggagg tactagacta cctttcatcc tacataaata gacgcatata agtacgcatt
     5641 taagccttgg cgccacttaa tgatttgcca gtaaaagaga ttgtttctag ctctcacatt
     5701 cttgcagata taatattgcc tagagctgaa gttatatatg attatcttaa gtaataaaaa
     5761 taagagttac cttaaatggt aactcttatt tttttaatgt cgactcatag aattcgtaat
     5821 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     5881 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     5941 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     6001 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     6061 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     6121 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     6181 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     6241 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     6301 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6361 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6421 aaagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6481 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6541 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6601 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6661 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6721 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     6781 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6841 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6901 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6961 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     7021 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     7081 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     7141 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     7201 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     7261 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     7321 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7381 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7441 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7501 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7561 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     7621 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7681 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7741 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7801 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7861 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7921 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     7981 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     8041 ttcgtc
//
