LOCUS       V015445                 5849 bp    DNA     circular SYN 30-JUN-2019
DEFINITION  Exported.
ACCESSION   V015445
VERSION     V015445
KEYWORDS    UNVERIFIED
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5849)
  AUTHORS   Lazarus JE, Warr AR, Kuehl CJ, Giorgio RT, Davis BM, Waldor MK.
  TITLE     A New Suite of Allelic Exchange Vectors for the Scarless
            Modification of Proteobacterial Genomes
  JOURNAL   Appl. Environ. Microbiol. (2019) In press
   PUBMED   31201277
REFERENCE   2  (bases 1 to 5849)
  AUTHORS   Lazarus JE, Warr AR, Kuehl CJ, Giorgio RT, Davis BM, Waldor MK.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-MAY-2019) Medicine, Massachusetts General Hospital, 55
            Fruit St, Jackson 5, Boston, MA 02114, USA
REFERENCE   3  (bases 1 to 5849)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
            Environ. Microbiol. (2019) In press"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (24-MAY-2019) Medicine, Massachusetts General Hospital, 55 Fruit St,
            Jackson 5, Boston, MA 02114, USA"
            GenBank staff is unable to verify sequence and/or annotation
            provided by the submitter.
FEATURES             Location/Qualifiers
     source          1..5849
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(4..534)
                     /label="GmR"
                     /note="gentamycin acetyltransferase"
     terminator      751..837
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      929..956
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        1159..1277
                     /label="rhaB promoter"
                     /note="promoter of the E. coli rhaBAD operon, conferring
                     tight induction with L-rhamnose and repression with
                     D-glucose in the presence of RhaR and RhaS (Giacalone et
                     al., 2006)"
     CDS             1301..1762
                     /gene="yhaV"
                     /label="Ribonuclease toxin YhaV"
                     /note="Ribonuclease toxin YhaV from Escherichia coli
                     (strain K12). Accession#: P64594"
     terminator      1825..2071
                     /label="lambda tL3 terminator"
                     /note="transcription terminator tL3 from phage lambda"
     protein_bind    complement(2107..2123)
                     /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)."
     rep_origin      complement(2158..2546)
                     /direction=LEFT
                     /label="R6K gamma ori"
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     CDS             complement(3167..3535)
                     /label="traJ"
                     /note="oriT-recognizing protein"
     oriT            complement(3568..3677)
                     /direction=LEFT
                     /label="oriT"
                     /note="incP origin of transfer"
     regulatory      4590..4607
                     /label="AmpR promoter"
                     /note="AmpR promoter"
                     /regulatory_class="promoter"
     regulatory      4608..4689
                     /label="AmpR promoter"
                     /note="AmpR promoter"
                     /regulatory_class="promoter"
     CDS             4717..5550
                     /label="rhaS"
                     /note="positive regulator of the rhaB promoter"
     regulatory      5638..5671
                     /label="strong forward terminator"
                     /note="strong forward terminator"
                     /regulatory_class="terminator"
ORIGIN
        1 ttaggtggcg gtacttgggt cgatatcaaa gtgcatcact tcttcccgta tgcccaactt
       61 tgtatagaga gccactgcgg gatcgtcacc gtaatctgct tgcacgtaga tcacataagc
      121 accaagcgcg ttggcctcat gcttgaggag attgatgagc gcggtggcaa tgccctgcct
      181 ccggtgctcg ccggagactg cgagatcata gatatagatc tcactacgcg gctgctcaaa
      241 cttgggcaga acgtaagccg cgagagcgcc aacaaccgct tcttggtcga aggcagcaag
      301 cgcgatgaat gtcttactac ggagcaagtt cccgaggtaa tcggagtccg gctgatgttg
      361 ggagtaggtg gctacgtctc cgaactcacg accgaaaaga tcaagagcag cccgcatgga
      421 tttgacttgg tcagggccga gcctacatgt gcgaatgatg cccatacttg agccacctaa
      481 ctttgtttta gggcgactgc cctgctgcgt aacatcgttg ctgctgcgta acatatgttt
      541 ttcctcctta tgttaagctt actcagttat tatatcataa atatctgtgt caagaataaa
      601 ctcccacatg gattcgacag tactgcgatg agtggcaggg cggggcgtaa tttttttaag
      661 gcagttattg tcttcaaatt cccgttgcat cgatggcccc cgatggtagt gtggggtctc
      721 cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac
      781 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
      841 ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg
      901 ccataaactg ccaggcatca aattaagcag aaggccatcc tgacggatgg cctttttgcg
      961 tggccagtgc caagcttgca tgcctgcagc tgtccggaac tagtcgaggc catggtggcc
     1021 tcctgatgtc gtcaacacgg cgaaatagta atcacgaggt caggttctta ccttaaattt
     1081 tcgacggaaa accacgtaaa aaacgtcgat ttttcaagat acagcgtgaa ttttcaggaa
     1141 atgcggtgag catcacatca ccacaattca gcaaattgtg aacatcatca cgttcatctt
     1201 tccctggttg ccaatggccc attttcctgt cagtaacgag aaggtcgcga attcaggcgc
     1261 tttttagact ggtcgtaatg aaattcagca ggatcaccat atggattttc cacaaagggt
     1321 taatggttgg gcgctatatg ctcatccctg ttttcaggaa acctacgacg ctttagttgc
     1381 cgaagtcgag acattaaagg gaaaagatcc tgaaaattat cagagaaaag ccgccacaaa
     1441 gttattggcg gtagtccata aagtgattga ggagcatatc acggtcaatc catcatcacc
     1501 ggcattccgt catggcaagt cgttaggctc tgggaaaaat aaagactggt cacgggtaaa
     1561 atttggtgct ggtcgttatc gtctcttctt tcgttatagt gaaaaagaga aagtcatcat
     1621 tctgggatgg atgaacgatg aaaacactct gcgcacctac ggtaaaaaaa cagatgccta
     1681 taccgtattc agcaaaatgt taaaaagagg acatcctcct gccgactggg aaaccctcac
     1741 ccgagaaaca gaagaaaccc attgaggatc cgcgaattct catgtttgac agcttatcac
     1801 tgatcagtga attaatggcg atgacgcatc ctcacgataa tatccgggta ggcgcaatca
     1861 ctttcgtctc tactccgtta caaagcgagg ctgggtattt cccggccttt ctgttatccg
     1921 aaatccactg aaagcacagc ggctggctga ggagataaat aataaacgag gggctgtatg
     1981 cacaaagcat cttctgttga gttaagaacg agtatcgaga tggcacatag ccttgctcaa
     2041 attggaatca ggtttgtgcc aataccagta gaaacagacg aagaagtcgt gggggaattc
     2101 cacaaattgt tatccgctca caattccaca tgtggaattc cacatgtgga attcccatgt
     2161 cagccgttaa gtgttcctgt gtcactcaaa attgctttga gaggctctaa gggcttctca
     2221 gtgcgttaca tccctggctt gttgtccaca accgttaaac cttaaaagct ttaaaagcct
     2281 tatatattct tttttttctt ataaaactta aaaccttaga ggctatttaa gttgctgatt
     2341 tatattaatt ttattgttca aacatgagag cttagtacgt gaaacatgag agcttagtac
     2401 gttagccatg agagcttagt acgttagcca tgagggttta gttcgttaaa catgagagct
     2461 tagtacgtta aacatgagag cttagtacgt gaaacatgag agcttagtac gtactatcaa
     2521 caggttgaac tgctgatctt cagatcctct acgccggacg catcgtggcc ggatccagcc
     2581 gaccaggctt tccacgcccg cgtgccgctc catgtcgttc gcgcggttct cggaaacgcg
     2641 ctgccgcgtt tcgtgattgt cacgctcaag cccgtagtcc cgttcgagcg tcgcgcagag
     2701 gtcagcgagg gcgcggtagg cccgatacgg ctcatggatg gtgtttcggg tcgggtgaat
     2761 cttgttgatg gcgatatgga tgtgcaggtt gtcggtgtcg tgatgcacgg cactgacgcg
     2821 ctgatgctcg gcgaagccaa gcccagcgca gatgcggtcc tcaatcgcgc gcaacgtctc
     2881 cgcgtcgggc ttctctcccg cgcggaagct aaccagcagg tgataggtct tgtcggcctc
     2941 ggaacgggtg ttgccgtgct gggtcgccat cacctcggcc atgacagcgg gcagggtgtt
     3001 tgcctcgcag ttcgtgacgc gcacgtgacc caggcgctcg gtcttgcctt gctcgtcggt
     3061 gatgtacttc accagctccg cgaagtcgct cttcttgatg gagcgcatgg ggacgtgctt
     3121 ggcaatcacg cgcacccccc ggccgtttta gcggctaaaa aagtcatggc tctgccctcg
     3181 ggcggaccac gcccatcatg accttgccaa gctcgtcctg cttctcttcg atcttcgcca
     3241 gcagggcgag gatcgtggca tcaccgaacc gcgccgtgcg cgggtcgtcg gtgagccaga
     3301 gtttcagcag gccgcccagg cggcccaggt cgccattgat gcgggccagc tcgcggacgt
     3361 gctcatagtc cacgacgccc gtgattttgt agccctggcc gacggccagc aggtaggccg
     3421 acaggctcat gccggccgcc gccgcctttt cctcaatcgc tcttcgttcg tctggaaggc
     3481 agtacacctt gataggtggg ctgcccttcc tggttggctt ggtttcatca gccatccgct
     3541 tgccctcatc tgttacgccg gcggtagccg gccagcctcg cagagcagga ttcccgttga
     3601 gcaccgccag gtgcgaataa gggacagtga agaaggaaca cccgctcgcg ggtgggccta
     3661 cttcacctat cctgcccggc tgacgccgtt ggatacacca aggaaagtct acacgaaccc
     3721 tttggcaaaa tcctgtatat cgtgcgaaaa aggatggata taccgaaaaa atcgctataa
     3781 tgaccccgaa gcagggttat gcagcggaaa agcgctgctt ccctgctgtt ttgtggaata
     3841 tctaccgact ggaaacaggc aaatgcagga aattactgaa ctgaggggac aggcgagaga
     3901 cgatgccaaa gagctacacc gacgagctgg ccgagtgggt tgaatcccgc gcggccaaga
     3961 agcgccggcg tgatgaggct gcggttgcgt tcctggcggt gagggcggat gtcgaggcgg
     4021 cgttagcgtc cggctatgcg ctcgtcacca tttgggagca catgcgggaa acggggaagg
     4081 tcaagttctc ctacgagacg ttccgctcgc acgccaggcg gcacatcaag gccaagcccg
     4141 ccgatgtgcc cgcaccgcag gccaaggctg cggaacccgc gccggcaccc aagacgccgg
     4201 agccacggcg gccgaagcag gggggcaagg ctgaaaagcc ggcccccgct gcggccccga
     4261 ccggcttcac cttcaaccca acaccggaca aaaaggatcc tctacgccgg acgcatcgtg
     4321 gccggcatca ccggcgccac aagtgcggtt gctggcgcct atatcgccga catcaccgat
     4381 ggggaagatc gggctcgcca cttcgggctc atgagcgctt gtttcggcgt gggtatggtg
     4441 gcaggccccg tggccggggg actgttgggc gccatctcct tgctgcctcg cgcgtttcgg
     4501 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     4561 agcggatgcc gtaagtaagg taaaggcctc gcggaacccc tatttgttta tttttctaaa
     4621 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4681 gaaaaaggag cttatcaccg ataaggaggt tttttaatga cggtgctgca ctcggttgac
     4741 ttcttcccta gcggcaatgc cagcgttgcc attgagccgc gcctgcctca agccgacttc
     4801 ccggagcacc accacgactt ccacgagatc gttatcgtgg agcacggtac cggcatccac
     4861 gttttcaacg gccaaccgta cacgattacg ggcggtaccg tgtgcttcgt tcgtgatcac
     4921 gaccgccact tatacgagca cacggacaac ttatgcttaa ccaacgtttt ataccgtagc
     4981 cctgaccgct tccaattcct ggcgggctta aaccaactgt taccgcagga attagacggc
     5041 caatacccta gccattggcg tgtgaatcat tcggtgctgc aacaagttcg ccaattagtg
     5101 gcgcaaatgg agcaacaaga gggcgagaac gacctgccga gcacggcgag ccgtgaaatt
     5161 ctgttcatgc agctgttact gctgttacgc aagtcgagcc tgcaagaaaa tttagagaat
     5221 tcggcgagcc gcctgaatct gctgttagcg tggttagaag atcacttcgc ggacgaagtt
     5281 aactgggatg cggttgccga ccagttcagc ctgagcttac gcaccctgca ccgccaactg
     5341 aaacaacaga ccggcttaac cccgcaacgc tatttaaacc gtttacgctt aatgaaggcg
     5401 cgccacttac tgcgccattc ggaagcgtcg gtgaccgata ttgcgtacca ctgtggcttt
     5461 tcggatagca atcatttcag caccctgttc cgtcgcgaat tcaattggag ccctcgcgac
     5521 atccgccaag gccgcgacgg tttcttacag tgaaggcctt ctagagtcga cctgcaggca
     5581 tgcaagcttg gctgttttgg cggatgagag ggtaccgcat gagcttcgag ctaatcgcgc
     5641 aaaaaacccc gcttcggcgg ggttttttcg ctgatcacgt acgatatctt cgaaccggtg
     5701 cacatggtgt acagggccct aggataggac gtcttaaggt ttaaaccagg ttaattaatt
     5761 taaatgcatc ccgggacgtc tcgagctcga tcggaccgcg gccgctagca cgtataccaa
     5821 gtgtcctgtt ttcttgccgc caaggatct
//
