LOCUS       V016266                 5681 bp    DNA     circular SYN 30-JUN-2019
DEFINITION  Exported.
ACCESSION   V016266
VERSION     V016266
KEYWORDS    UNVERIFIED
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5681)
  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 5681)
  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 5681)
  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..5681
                     /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..1594
                     /gene="mqsR"
                     /label="mRNA interferase toxin MqsR"
                     /note="mRNA interferase toxin MqsR from Escherichia coli
                     (strain K12). Accession#: Q46865"
     terminator      1657..1903
                     /label="lambda tL3 terminator"
                     /note="transcription terminator tL3 from phage lambda"
     protein_bind    complement(1939..1955)
                     /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(1990..2378)
                     /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(2999..3367)
                     /label="traJ"
                     /note="oriT-recognizing protein"
     oriT            complement(3400..3509)
                     /direction=LEFT
                     /label="oriT"
                     /note="incP origin of transfer"
     regulatory      4422..4439
                     /label="AmpR promoter"
                     /note="AmpR promoter"
                     /regulatory_class="promoter"
     regulatory      4440..4521
                     /label="AmpR promoter"
                     /note="AmpR promoter"
                     /regulatory_class="promoter"
     CDS             4549..5382
                     /label="rhaS"
                     /note="positive regulator of the rhaB promoter"
     regulatory      5470..5503
                     /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 atggaaaaac gcacaccaca
     1321 tacacgtttg agtcaggtta aaaaacttgt caatgccggg caagttcgta caacacgtag
     1381 tgccctgtta aatgcagatg agttaggttt ggattttgat ggtatgtgta atgttatcat
     1441 tggattatca gagagcgact tttataaaag catgaccacc tactctgatc atactatctg
     1501 gcaggatgtt tacagaccca ggcttgttac aggccaggtt tatcttaaaa ttacggtaat
     1561 tcatgacgta ctgatcgtct cgtttaagga gaagtaagga tccgcgaatt ctcatgtttg
     1621 acagcttatc actgatcagt gaattaatgg cgatgacgca tcctcacgat aatatccggg
     1681 taggcgcaat cactttcgtc tctactccgt tacaaagcga ggctgggtat ttcccggcct
     1741 ttctgttatc cgaaatccac tgaaagcaca gcggctggct gaggagataa ataataaacg
     1801 aggggctgta tgcacaaagc atcttctgtt gagttaagaa cgagtatcga gatggcacat
     1861 agccttgctc aaattggaat caggtttgtg ccaataccag tagaaacaga cgaagaagtc
     1921 gtgggggaat tccacaaatt gttatccgct cacaattcca catgtggaat tccacatgtg
     1981 gaattcccat gtcagccgtt aagtgttcct gtgtcactca aaattgcttt gagaggctct
     2041 aagggcttct cagtgcgtta catccctggc ttgttgtcca caaccgttaa accttaaaag
     2101 ctttaaaagc cttatatatt cttttttttc ttataaaact taaaacctta gaggctattt
     2161 aagttgctga tttatattaa ttttattgtt caaacatgag agcttagtac gtgaaacatg
     2221 agagcttagt acgttagcca tgagagctta gtacgttagc catgagggtt tagttcgtta
     2281 aacatgagag cttagtacgt taaacatgag agcttagtac gtgaaacatg agagcttagt
     2341 acgtactatc aacaggttga actgctgatc ttcagatcct ctacgccgga cgcatcgtgg
     2401 ccggatccag ccgaccaggc tttccacgcc cgcgtgccgc tccatgtcgt tcgcgcggtt
     2461 ctcggaaacg cgctgccgcg tttcgtgatt gtcacgctca agcccgtagt cccgttcgag
     2521 cgtcgcgcag aggtcagcga gggcgcggta ggcccgatac ggctcatgga tggtgtttcg
     2581 ggtcgggtga atcttgttga tggcgatatg gatgtgcagg ttgtcggtgt cgtgatgcac
     2641 ggcactgacg cgctgatgct cggcgaagcc aagcccagcg cagatgcggt cctcaatcgc
     2701 gcgcaacgtc tccgcgtcgg gcttctctcc cgcgcggaag ctaaccagca ggtgataggt
     2761 cttgtcggcc tcggaacggg tgttgccgtg ctgggtcgcc atcacctcgg ccatgacagc
     2821 gggcagggtg tttgcctcgc agttcgtgac gcgcacgtga cccaggcgct cggtcttgcc
     2881 ttgctcgtcg gtgatgtact tcaccagctc cgcgaagtcg ctcttcttga tggagcgcat
     2941 ggggacgtgc ttggcaatca cgcgcacccc ccggccgttt tagcggctaa aaaagtcatg
     3001 gctctgccct cgggcggacc acgcccatca tgaccttgcc aagctcgtcc tgcttctctt
     3061 cgatcttcgc cagcagggcg aggatcgtgg catcaccgaa ccgcgccgtg cgcgggtcgt
     3121 cggtgagcca gagtttcagc aggccgccca ggcggcccag gtcgccattg atgcgggcca
     3181 gctcgcggac gtgctcatag tccacgacgc ccgtgatttt gtagccctgg ccgacggcca
     3241 gcaggtaggc cgacaggctc atgccggccg ccgccgcctt ttcctcaatc gctcttcgtt
     3301 cgtctggaag gcagtacacc ttgataggtg ggctgccctt cctggttggc ttggtttcat
     3361 cagccatccg cttgccctca tctgttacgc cggcggtagc cggccagcct cgcagagcag
     3421 gattcccgtt gagcaccgcc aggtgcgaat aagggacagt gaagaaggaa cacccgctcg
     3481 cgggtgggcc tacttcacct atcctgcccg gctgacgccg ttggatacac caaggaaagt
     3541 ctacacgaac cctttggcaa aatcctgtat atcgtgcgaa aaaggatgga tataccgaaa
     3601 aaatcgctat aatgaccccg aagcagggtt atgcagcgga aaagcgctgc ttccctgctg
     3661 ttttgtggaa tatctaccga ctggaaacag gcaaatgcag gaaattactg aactgagggg
     3721 acaggcgaga gacgatgcca aagagctaca ccgacgagct ggccgagtgg gttgaatccc
     3781 gcgcggccaa gaagcgccgg cgtgatgagg ctgcggttgc gttcctggcg gtgagggcgg
     3841 atgtcgaggc ggcgttagcg tccggctatg cgctcgtcac catttgggag cacatgcggg
     3901 aaacggggaa ggtcaagttc tcctacgaga cgttccgctc gcacgccagg cggcacatca
     3961 aggccaagcc cgccgatgtg cccgcaccgc aggccaaggc tgcggaaccc gcgccggcac
     4021 ccaagacgcc ggagccacgg cggccgaagc aggggggcaa ggctgaaaag ccggcccccg
     4081 ctgcggcccc gaccggcttc accttcaacc caacaccgga caaaaaggat cctctacgcc
     4141 ggacgcatcg tggccggcat caccggcgcc acaagtgcgg ttgctggcgc ctatatcgcc
     4201 gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc
     4261 gtgggtatgg tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgctgcct
     4321 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     4381 agcttgtctg taagcggatg ccgtaagtaa ggtaaaggcc tcgcggaacc cctatttgtt
     4441 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     4501 ttcaataata ttgaaaaagg agcttatcac cgataaggag gttttttaat gacggtgctg
     4561 cactcggttg acttcttccc tagcggcaat gccagcgttg ccattgagcc gcgcctgcct
     4621 caagccgact tcccggagca ccaccacgac ttccacgaga tcgttatcgt ggagcacggt
     4681 accggcatcc acgttttcaa cggccaaccg tacacgatta cgggcggtac cgtgtgcttc
     4741 gttcgtgatc acgaccgcca cttatacgag cacacggaca acttatgctt aaccaacgtt
     4801 ttataccgta gccctgaccg cttccaattc ctggcgggct taaaccaact gttaccgcag
     4861 gaattagacg gccaataccc tagccattgg cgtgtgaatc attcggtgct gcaacaagtt
     4921 cgccaattag tggcgcaaat ggagcaacaa gagggcgaga acgacctgcc gagcacggcg
     4981 agccgtgaaa ttctgttcat gcagctgtta ctgctgttac gcaagtcgag cctgcaagaa
     5041 aatttagaga attcggcgag ccgcctgaat ctgctgttag cgtggttaga agatcacttc
     5101 gcggacgaag ttaactggga tgcggttgcc gaccagttca gcctgagctt acgcaccctg
     5161 caccgccaac tgaaacaaca gaccggctta accccgcaac gctatttaaa ccgtttacgc
     5221 ttaatgaagg cgcgccactt actgcgccat tcggaagcgt cggtgaccga tattgcgtac
     5281 cactgtggct tttcggatag caatcatttc agcaccctgt tccgtcgcga attcaattgg
     5341 agccctcgcg acatccgcca aggccgcgac ggtttcttac agtgaaggcc ttctagagtc
     5401 gacctgcagg catgcaagct tggctgtttt ggcggatgag agggtaccgc atgagcttcg
     5461 agctaatcgc gcaaaaaacc ccgcttcggc ggggtttttt cgctgatcac gtacgatatc
     5521 ttcgaaccgg tgcacatggt gtacagggcc ctaggatagg acgtcttaag gtttaaacca
     5581 ggttaattaa tttaaatgca tcccgggacg tctcgagctc gatcggaccg cggccgctag
     5641 cacgtatacc aagtgtcctg ttttcttgcc gccaaggatc t
//
