LOCUS       V016205                 5878 bp    DNA     circular SYN 30-JUN-2019
DEFINITION  Exported.
ACCESSION   V016205
VERSION     V016205
KEYWORDS    UNVERIFIED
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5878)
  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 5878)
  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 5878)
  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..5878
                     /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             1318..1791
                     /gene="tse2"
                     /label="Toxin Tse2"
                     /note="Toxin Tse2 from Pseudomonas aeruginosa (strain ATCC
                     15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C
                     / PRS 101 / PAO1). Accession#: Q9I0E0"
     terminator      1854..2100
                     /label="lambda tL3 terminator"
                     /note="transcription terminator tL3 from phage lambda"
     protein_bind    complement(2136..2152)
                     /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(2187..2575)
                     /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(3196..3564)
                     /label="traJ"
                     /note="oriT-recognizing protein"
     oriT            complement(3597..3706)
                     /direction=LEFT
                     /label="oriT"
                     /note="incP origin of transfer"
     regulatory      4619..4636
                     /label="AmpR promoter"
                     /note="AmpR promoter"
                     /regulatory_class="promoter"
     regulatory      4637..4718
                     /label="AmpR promoter"
                     /note="AmpR promoter"
                     /regulatory_class="promoter"
     CDS             4746..5579
                     /label="rhaS"
                     /note="positive regulator of the rhaB promoter"
     regulatory      5667..5700
                     /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 atgaggaggt aaaatagatg
     1321 tcctacgact acgagaaaac cagcctcacc ctctaccggg cggtattcaa ggccaactac
     1381 gacggcgacg tcggtcgcta cctgcatccc gacaaggaac tcgccgaggc tgcggaagtc
     1441 gccccgctgc tgcatccgac cttcgacagc cccaacaccc ctggcgtccc cgcccgcgcg
     1501 ccggacatcg tcgccggccg cgacggcctc tacgccccgg acaccggcgg cacctcggtg
     1561 ttcgaccgcg ccggcgtgct gcgccgcgcc gacggcgact tcgtgatacc cgacggcacc
     1621 gacatcccgc cggaccttaa ggtgaagcag gacagctaca acaagcgcct gcaagccacc
     1681 cactacacca tcatgccggc caagccgatg taccgggagg tcctcatggg ccaactggac
     1741 aacttcgtgc gcaacgccat ccgccgccaa tgggaaaaag cccgcgggct ctagggatcc
     1801 gcgaattctc atgtttgaca gcttatcact gatcagtgaa ttaatggcga tgacgcatcc
     1861 tcacgataat atccgggtag gcgcaatcac tttcgtctct actccgttac aaagcgaggc
     1921 tgggtatttc ccggcctttc tgttatccga aatccactga aagcacagcg gctggctgag
     1981 gagataaata ataaacgagg ggctgtatgc acaaagcatc ttctgttgag ttaagaacga
     2041 gtatcgagat ggcacatagc cttgctcaaa ttggaatcag gtttgtgcca ataccagtag
     2101 aaacagacga agaagtcgtg ggggaattcc acaaattgtt atccgctcac aattccacat
     2161 gtggaattcc acatgtggaa ttcccatgtc agccgttaag tgttcctgtg tcactcaaaa
     2221 ttgctttgag aggctctaag ggcttctcag tgcgttacat ccctggcttg ttgtccacaa
     2281 ccgttaaacc ttaaaagctt taaaagcctt atatattctt ttttttctta taaaacttaa
     2341 aaccttagag gctatttaag ttgctgattt atattaattt tattgttcaa acatgagagc
     2401 ttagtacgtg aaacatgaga gcttagtacg ttagccatga gagcttagta cgttagccat
     2461 gagggtttag ttcgttaaac atgagagctt agtacgttaa acatgagagc ttagtacgtg
     2521 aaacatgaga gcttagtacg tactatcaac aggttgaact gctgatcttc agatcctcta
     2581 cgccggacgc atcgtggccg gatccagccg accaggcttt ccacgcccgc gtgccgctcc
     2641 atgtcgttcg cgcggttctc ggaaacgcgc tgccgcgttt cgtgattgtc acgctcaagc
     2701 ccgtagtccc gttcgagcgt cgcgcagagg tcagcgaggg cgcggtaggc ccgatacggc
     2761 tcatggatgg tgtttcgggt cgggtgaatc ttgttgatgg cgatatggat gtgcaggttg
     2821 tcggtgtcgt gatgcacggc actgacgcgc tgatgctcgg cgaagccaag cccagcgcag
     2881 atgcggtcct caatcgcgcg caacgtctcc gcgtcgggct tctctcccgc gcggaagcta
     2941 accagcaggt gataggtctt gtcggcctcg gaacgggtgt tgccgtgctg ggtcgccatc
     3001 acctcggcca tgacagcggg cagggtgttt gcctcgcagt tcgtgacgcg cacgtgaccc
     3061 aggcgctcgg tcttgccttg ctcgtcggtg atgtacttca ccagctccgc gaagtcgctc
     3121 ttcttgatgg agcgcatggg gacgtgcttg gcaatcacgc gcaccccccg gccgttttag
     3181 cggctaaaaa agtcatggct ctgccctcgg gcggaccacg cccatcatga ccttgccaag
     3241 ctcgtcctgc ttctcttcga tcttcgccag cagggcgagg atcgtggcat caccgaaccg
     3301 cgccgtgcgc gggtcgtcgg tgagccagag tttcagcagg ccgcccaggc ggcccaggtc
     3361 gccattgatg cgggccagct cgcggacgtg ctcatagtcc acgacgcccg tgattttgta
     3421 gccctggccg acggccagca ggtaggccga caggctcatg ccggccgccg ccgccttttc
     3481 ctcaatcgct cttcgttcgt ctggaaggca gtacaccttg ataggtgggc tgcccttcct
     3541 ggttggcttg gtttcatcag ccatccgctt gccctcatct gttacgccgg cggtagccgg
     3601 ccagcctcgc agagcaggat tcccgttgag caccgccagg tgcgaataag ggacagtgaa
     3661 gaaggaacac ccgctcgcgg gtgggcctac ttcacctatc ctgcccggct gacgccgttg
     3721 gatacaccaa ggaaagtcta cacgaaccct ttggcaaaat cctgtatatc gtgcgaaaaa
     3781 ggatggatat accgaaaaaa tcgctataat gaccccgaag cagggttatg cagcggaaaa
     3841 gcgctgcttc cctgctgttt tgtggaatat ctaccgactg gaaacaggca aatgcaggaa
     3901 attactgaac tgaggggaca ggcgagagac gatgccaaag agctacaccg acgagctggc
     3961 cgagtgggtt gaatcccgcg cggccaagaa gcgccggcgt gatgaggctg cggttgcgtt
     4021 cctggcggtg agggcggatg tcgaggcggc gttagcgtcc ggctatgcgc tcgtcaccat
     4081 ttgggagcac atgcgggaaa cggggaaggt caagttctcc tacgagacgt tccgctcgca
     4141 cgccaggcgg cacatcaagg ccaagcccgc cgatgtgccc gcaccgcagg ccaaggctgc
     4201 ggaacccgcg ccggcaccca agacgccgga gccacggcgg ccgaagcagg ggggcaaggc
     4261 tgaaaagccg gcccccgctg cggccccgac cggcttcacc ttcaacccaa caccggacaa
     4321 aaaggatcct ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca agtgcggttg
     4381 ctggcgccta tatcgccgac atcaccgatg gggaagatcg ggctcgccac ttcgggctca
     4441 tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt ggccggggga ctgttgggcg
     4501 ccatctcctt gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     4561 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg taagtaaggt aaaggcctcg
     4621 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4681 ataaccctga taaatgcttc aataatattg aaaaaggagc ttatcaccga taaggaggtt
     4741 ttttaatgac ggtgctgcac tcggttgact tcttccctag cggcaatgcc agcgttgcca
     4801 ttgagccgcg cctgcctcaa gccgacttcc cggagcacca ccacgacttc cacgagatcg
     4861 ttatcgtgga gcacggtacc ggcatccacg ttttcaacgg ccaaccgtac acgattacgg
     4921 gcggtaccgt gtgcttcgtt cgtgatcacg accgccactt atacgagcac acggacaact
     4981 tatgcttaac caacgtttta taccgtagcc ctgaccgctt ccaattcctg gcgggcttaa
     5041 accaactgtt accgcaggaa ttagacggcc aataccctag ccattggcgt gtgaatcatt
     5101 cggtgctgca acaagttcgc caattagtgg cgcaaatgga gcaacaagag ggcgagaacg
     5161 acctgccgag cacggcgagc cgtgaaattc tgttcatgca gctgttactg ctgttacgca
     5221 agtcgagcct gcaagaaaat ttagagaatt cggcgagccg cctgaatctg ctgttagcgt
     5281 ggttagaaga tcacttcgcg gacgaagtta actgggatgc ggttgccgac cagttcagcc
     5341 tgagcttacg caccctgcac cgccaactga aacaacagac cggcttaacc ccgcaacgct
     5401 atttaaaccg tttacgctta atgaaggcgc gccacttact gcgccattcg gaagcgtcgg
     5461 tgaccgatat tgcgtaccac tgtggctttt cggatagcaa tcatttcagc accctgttcc
     5521 gtcgcgaatt caattggagc cctcgcgaca tccgccaagg ccgcgacggt ttcttacagt
     5581 gaaggccttc tagagtcgac ctgcaggcat gcaagcttgg ctgttttggc ggatgagagg
     5641 gtaccgcatg agcttcgagc taatcgcgca aaaaaccccg cttcggcggg gttttttcgc
     5701 tgatcacgta cgatatcttc gaaccggtgc acatggtgta cagggcccta ggataggacg
     5761 tcttaaggtt taaaccaggt taattaattt aaatgcatcc cgggacgtct cgagctcgat
     5821 cggaccgcgg ccgctagcac gtataccaag tgtcctgttt tcttgccgcc aaggatct
//
