LOCUS       V016101                 6640 bp    DNA     circular SYN 30-JUN-2019
DEFINITION  Exported.
ACCESSION   V016101
VERSION     V016101
KEYWORDS    UNVERIFIED
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6640)
  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 6640)
  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 6640)
  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..6640
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      62..95
                     /label="strong forward terminator"
                     /note="strong forward terminator"
                     /regulatory_class="terminator"
     promoter        346..448
                     /label="cat promoter"
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             449..1105
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     terminator      1209..1295
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      1387..1414
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        1617..1735
                     /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             1759..2052
                     /gene="mqsR"
                     /label="mRNA interferase toxin MqsR"
                     /note="mRNA interferase toxin MqsR from Escherichia coli
                     (strain K12). Accession#: Q46865"
     terminator      2115..2361
                     /label="lambda tL3 terminator"
                     /note="transcription terminator tL3 from phage lambda"
     protein_bind    complement(2397..2413)
                     /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(2448..2836)
                     /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(3457..3825)
                     /label="traJ"
                     /note="oriT-recognizing protein"
     oriT            complement(3858..3967)
                     /direction=LEFT
                     /label="oriT"
                     /note="incP origin of transfer"
     regulatory      4880..4897
                     /label="AmpR promoter"
                     /note="AmpR promoter"
                     /regulatory_class="promoter"
     regulatory      4898..4979
                     /label="AmpR promoter"
                     /note="AmpR promoter"
                     /regulatory_class="promoter"
     CDS             5007..5840
                     /label="rhaS"
                     /note="positive regulator of the rhaB promoter"
     promoter        5878..5906
                     /label="tac promoter"
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     CDS             5973..6635
                     /label="amilCP"
                     /note="amilCP is a fluorescent protein published in 2008,
                     derived from Acropora millepora. It is reported to be a
                     somewhat slowly-maturing tetramer."
ORIGIN
        1 ggcatgcaag cttggctgtt ttggcggatg agagggtacc gcatgagctt cgagctaatc
       61 gcgcaaaaaa ccccgcttcg gcggggtttt ttcgctgatc acgtacgata tcttcgaacc
      121 ggtgcacatg gtgtacaggg ccctaggata ggacgtctta aggtttaaac caggttaatt
      181 aatttaaatg catcccggga cgtctcgagc tcgatcggac cgcggccgct agcacgtata
      241 ccaagtgtcc tgttttcttg ccgccaagga tctgatggcg caggggatca agatctctgt
      301 tgataccggg aagccctggg ccaacttttg gcgaaaatga gacgttgatc ggcacgtaag
      361 aggttccaac tttcaccata atgaaataag atcactaccg ggcgtatttt ttgagttatc
      421 gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt
      481 tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg
      541 tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa
      601 taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc
      661 ggaattacgt atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg
      721 ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga
      781 cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct
      841 ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt
      901 gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt
      961 caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt
     1021 tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta
     1081 ctgcgatgag tggcagggcg gggcgtaatt tttttaaggc agttattgtc ttcaaattcc
     1141 cgttgcatcg atggcccccg atggtagtgt ggggtctccc catgcgagag tagggaactg
     1201 ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
     1261 gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg
     1321 cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa
     1381 ttaagcagaa ggccatcctg acggatggcc tttttgcgtg gccagtgcca agcttgcatg
     1441 cctgcagctg tccggaacta gtcgaggcca tggtggcctc ctgatgtcgt caacacggcg
     1501 aaatagtaat cacgaggtca ggttcttacc ttaaattttc gacggaaaac cacgtaaaaa
     1561 acgtcgattt ttcaagatac agcgtgaatt ttcaggaaat gcggtgagca tcacatcacc
     1621 acaattcagc aaattgtgaa catcatcacg ttcatctttc cctggttgcc aatggcccat
     1681 tttcctgtca gtaacgagaa ggtcgcgaat tcaggcgctt tttagactgg tcgtaatgaa
     1741 attcagcagg atcaccatat ggaaaaacgc acaccacata cacgtttgag tcaggttaaa
     1801 aaacttgtca atgccgggca agttcgtaca acacgtagtg ccctgttaaa tgcagatgag
     1861 ttaggtttgg attttgatgg tatgtgtaat gttatcattg gattatcaga gagcgacttt
     1921 tataaaagca tgaccaccta ctctgatcat actatctggc aggatgttta cagacccagg
     1981 cttgttacag gccaggttta tcttaaaatt acggtaattc atgacgtact gatcgtctcg
     2041 tttaaggaga agtaaggatc cgcgaattct catgtttgac agcttatcac tgatcagtga
     2101 attaatggcg atgacgcatc ctcacgataa tatccgggta ggcgcaatca ctttcgtctc
     2161 tactccgtta caaagcgagg ctgggtattt cccggccttt ctgttatccg aaatccactg
     2221 aaagcacagc ggctggctga ggagataaat aataaacgag gggctgtatg cacaaagcat
     2281 cttctgttga gttaagaacg agtatcgaga tggcacatag ccttgctcaa attggaatca
     2341 ggtttgtgcc aataccagta gaaacagacg aagaagtcgt gggggaattc cacaaattgt
     2401 tatccgctca caattccaca tgtggaattc cacatgtgga attcccatgt cagccgttaa
     2461 gtgttcctgt gtcactcaaa attgctttga gaggctctaa gggcttctca gtgcgttaca
     2521 tccctggctt gttgtccaca accgttaaac cttaaaagct ttaaaagcct tatatattct
     2581 tttttttctt ataaaactta aaaccttaga ggctatttaa gttgctgatt tatattaatt
     2641 ttattgttca aacatgagag cttagtacgt gaaacatgag agcttagtac gttagccatg
     2701 agagcttagt acgttagcca tgagggttta gttcgttaaa catgagagct tagtacgtta
     2761 aacatgagag cttagtacgt gaaacatgag agcttagtac gtactatcaa caggttgaac
     2821 tgctgatctt cagatcctct acgccggacg catcgtggcc ggatccagcc gaccaggctt
     2881 tccacgcccg cgtgccgctc catgtcgttc gcgcggttct cggaaacgcg ctgccgcgtt
     2941 tcgtgattgt cacgctcaag cccgtagtcc cgttcgagcg tcgcgcagag gtcagcgagg
     3001 gcgcggtagg cccgatacgg ctcatggatg gtgtttcggg tcgggtgaat cttgttgatg
     3061 gcgatatgga tgtgcaggtt gtcggtgtcg tgatgcacgg cactgacgcg ctgatgctcg
     3121 gcgaagccaa gcccagcgca gatgcggtcc tcaatcgcgc gcaacgtctc cgcgtcgggc
     3181 ttctctcccg cgcggaagct aaccagcagg tgataggtct tgtcggcctc ggaacgggtg
     3241 ttgccgtgct gggtcgccat cacctcggcc atgacagcgg gcagggtgtt tgcctcgcag
     3301 ttcgtgacgc gcacgtgacc caggcgctcg gtcttgcctt gctcgtcggt gatgtacttc
     3361 accagctccg cgaagtcgct cttcttgatg gagcgcatgg ggacgtgctt ggcaatcacg
     3421 cgcacccccc ggccgtttta gcggctaaaa aagtcatggc tctgccctcg ggcggaccac
     3481 gcccatcatg accttgccaa gctcgtcctg cttctcttcg atcttcgcca gcagggcgag
     3541 gatcgtggca tcaccgaacc gcgccgtgcg cgggtcgtcg gtgagccaga gtttcagcag
     3601 gccgcccagg cggcccaggt cgccattgat gcgggccagc tcgcggacgt gctcatagtc
     3661 cacgacgccc gtgattttgt agccctggcc gacggccagc aggtaggccg acaggctcat
     3721 gccggccgcc gccgcctttt cctcaatcgc tcttcgttcg tctggaaggc agtacacctt
     3781 gataggtggg ctgcccttcc tggttggctt ggtttcatca gccatccgct tgccctcatc
     3841 tgttacgccg gcggtagccg gccagcctcg cagagcagga ttcccgttga gcaccgccag
     3901 gtgcgaataa gggacagtga agaaggaaca cccgctcgcg ggtgggccta cttcacctat
     3961 cctgcccggc tgacgccgtt ggatacacca aggaaagtct acacgaaccc tttggcaaaa
     4021 tcctgtatat cgtgcgaaaa aggatggata taccgaaaaa atcgctataa tgaccccgaa
     4081 gcagggttat gcagcggaaa agcgctgctt ccctgctgtt ttgtggaata tctaccgact
     4141 ggaaacaggc aaatgcagga aattactgaa ctgaggggac aggcgagaga cgatgccaaa
     4201 gagctacacc gacgagctgg ccgagtgggt tgaatcccgc gcggccaaga agcgccggcg
     4261 tgatgaggct gcggttgcgt tcctggcggt gagggcggat gtcgaggcgg cgttagcgtc
     4321 cggctatgcg ctcgtcacca tttgggagca catgcgggaa acggggaagg tcaagttctc
     4381 ctacgagacg ttccgctcgc acgccaggcg gcacatcaag gccaagcccg ccgatgtgcc
     4441 cgcaccgcag gccaaggctg cggaacccgc gccggcaccc aagacgccgg agccacggcg
     4501 gccgaagcag gggggcaagg ctgaaaagcc ggcccccgct gcggccccga ccggcttcac
     4561 cttcaaccca acaccggaca aaaaggatcc tctacgccgg acgcatcgtg gccggcatca
     4621 ccggcgccac aagtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc
     4681 gggctcgcca cttcgggctc atgagcgctt gtttcggcgt gggtatggtg gcaggccccg
     4741 tggccggggg actgttgggc gccatctcct tgctgcctcg cgcgtttcgg tgatgacggt
     4801 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     4861 gtaagtaagg taaaggcctc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4921 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggag
     4981 cttatcaccg ataaggaggt tttttaatga cggtgctgca ctcggttgac ttcttcccta
     5041 gcggcaatgc cagcgttgcc attgagccgc gcctgcctca agccgacttc ccggagcacc
     5101 accacgactt ccacgagatc gttatcgtgg agcacggtac cggcatccac gttttcaacg
     5161 gccaaccgta cacgattacg ggcggtaccg tgtgcttcgt tcgtgatcac gaccgccact
     5221 tatacgagca cacggacaac ttatgcttaa ccaacgtttt ataccgtagc cctgaccgct
     5281 tccaattcct ggcgggctta aaccaactgt taccgcagga attagacggc caatacccta
     5341 gccattggcg tgtgaatcat tcggtgctgc aacaagttcg ccaattagtg gcgcaaatgg
     5401 agcaacaaga gggcgagaac gacctgccga gcacggcgag ccgtgaaatt ctgttcatgc
     5461 agctgttact gctgttacgc aagtcgagcc tgcaagaaaa tttagagaat tcggcgagcc
     5521 gcctgaatct gctgttagcg tggttagaag atcacttcgc ggacgaagtt aactgggatg
     5581 cggttgccga ccagttcagc ctgagcttac gcaccctgca ccgccaactg aaacaacaga
     5641 ccggcttaac cccgcaacgc tatttaaacc gtttacgctt aatgaaggcg cgccacttac
     5701 tgcgccattc ggaagcgtcg gtgaccgata ttgcgtacca ctgtggcttt tcggatagca
     5761 atcatttcag caccctgttc cgtcgcgaat tcaattggag ccctcgcgac atccgccaag
     5821 gccgcgacgg tttcttacag tgaaggcctt ctagagtcga cctgcatcga cctgcagttg
     5881 acaattaatc atcggctcgt ataatgtgtg gaaaggcggt tcaccgccgt tttcacacag
     5941 gaaacagaat tctttaagaa ggagatatac atatgtcagt gatagcaaag cagatgacat
     6001 acaaagtata tatgagcggt actgtaaatg gtcactactt cgaagtagaa ggtgatggca
     6061 aagggaagcc gtatgagggt gaacaaacag tgaagcttac agttacgaag ggtggccctt
     6121 tgccgttcgc gtgggacatt ttgtcgccac agtgccagta cgggagcata ccattcacca
     6181 agtatccaga ggacatacca gactacgtga aacagtcctt tcccgaaggc tatacctggg
     6241 agcgcataat gaactttgaa gacggtgcgg tttgtacggt atcgaacgat tcatcaatcc
     6301 aaggaaattg ctttatttat catgttaaat tttcggggct taactttccg ccaaatggcc
     6361 ccgtgatgca gaagaaaact cagggctggg aaccaaatac cgagcgcctt ttcgctcggg
     6421 acggaatgct gttgggaaac aattttatgg cgttgaagct ggaagggggt ggccactatc
     6481 tctgtgaatt caagacaaca tacaaagcca agaagcctgt caagatgccc ggttatcact
     6541 atgtggaccg gaagctcgat gtcactaatc acaacaagga ttatacttca gttgaacagt
     6601 gcgaaatttc gatcgctcgc aaacctgtcg tagcataagg
//
