LOCUS       V015422                 6063 bp    DNA     circular SYN 30-JUN-2019
DEFINITION  Exported.
ACCESSION   V015422
VERSION     V015422
KEYWORDS    UNVERIFIED
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6063)
  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 6063)
  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 6063)
  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..6063
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(4..392)
                     /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(1013..1381)
                     /label="traJ"
                     /note="oriT-recognizing protein"
     oriT            complement(1414..1523)
                     /direction=LEFT
                     /label="oriT"
                     /note="incP origin of transfer"
     regulatory      2436..2453
                     /label="AmpR promoter"
                     /note="AmpR promoter"
                     /regulatory_class="promoter"
     regulatory      2454..2535
                     /label="AmpR promoter"
                     /note="AmpR promoter"
                     /regulatory_class="promoter"
     CDS             2563..3396
                     /label="rhaS"
                     /note="positive regulator of the rhaB promoter"
     regulatory      3484..3517
                     /label="strong forward terminator"
                     /note="strong forward terminator"
                     /regulatory_class="terminator"
     promoter        3768..3870
                     /label="cat promoter"
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             3871..4527
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     terminator      4631..4717
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      4809..4836
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        5039..5157
                     /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             5198..5671
                     /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      5734..5980
                     /label="lambda tL3 terminator"
                     /note="transcription terminator tL3 from phage lambda"
     protein_bind    complement(6016..6032)
                     /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)."
ORIGIN
        1 ccatgtcagc cgttaagtgt tcctgtgtca ctcaaaattg ctttgagagg ctctaagggc
       61 ttctcagtgc gttacatccc tggcttgttg tccacaaccg ttaaacctta aaagctttaa
      121 aagccttata tattcttttt tttcttataa aacttaaaac cttagaggct atttaagttg
      181 ctgatttata ttaattttat tgttcaaaca tgagagctta gtacgtgaaa catgagagct
      241 tagtacgtta gccatgagag cttagtacgt tagccatgag ggtttagttc gttaaacatg
      301 agagcttagt acgttaaaca tgagagctta gtacgtgaaa catgagagct tagtacgtac
      361 tatcaacagg ttgaactgct gatcttcaga tcctctacgc cggacgcatc gtggccggat
      421 ccagccgacc aggctttcca cgcccgcgtg ccgctccatg tcgttcgcgc ggttctcgga
      481 aacgcgctgc cgcgtttcgt gattgtcacg ctcaagcccg tagtcccgtt cgagcgtcgc
      541 gcagaggtca gcgagggcgc ggtaggcccg atacggctca tggatggtgt ttcgggtcgg
      601 gtgaatcttg ttgatggcga tatggatgtg caggttgtcg gtgtcgtgat gcacggcact
      661 gacgcgctga tgctcggcga agccaagccc agcgcagatg cggtcctcaa tcgcgcgcaa
      721 cgtctccgcg tcgggcttct ctcccgcgcg gaagctaacc agcaggtgat aggtcttgtc
      781 ggcctcggaa cgggtgttgc cgtgctgggt cgccatcacc tcggccatga cagcgggcag
      841 ggtgtttgcc tcgcagttcg tgacgcgcac gtgacccagg cgctcggtct tgccttgctc
      901 gtcggtgatg tacttcacca gctccgcgaa gtcgctcttc ttgatggagc gcatggggac
      961 gtgcttggca atcacgcgca ccccccggcc gttttagcgg ctaaaaaagt catggctctg
     1021 ccctcgggcg gaccacgccc atcatgacct tgccaagctc gtcctgcttc tcttcgatct
     1081 tcgccagcag ggcgaggatc gtggcatcac cgaaccgcgc cgtgcgcggg tcgtcggtga
     1141 gccagagttt cagcaggccg cccaggcggc ccaggtcgcc attgatgcgg gccagctcgc
     1201 ggacgtgctc atagtccacg acgcccgtga ttttgtagcc ctggccgacg gccagcaggt
     1261 aggccgacag gctcatgccg gccgccgccg ccttttcctc aatcgctctt cgttcgtctg
     1321 gaaggcagta caccttgata ggtgggctgc ccttcctggt tggcttggtt tcatcagcca
     1381 tccgcttgcc ctcatctgtt acgccggcgg tagccggcca gcctcgcaga gcaggattcc
     1441 cgttgagcac cgccaggtgc gaataaggga cagtgaagaa ggaacacccg ctcgcgggtg
     1501 ggcctacttc acctatcctg cccggctgac gccgttggat acaccaagga aagtctacac
     1561 gaaccctttg gcaaaatcct gtatatcgtg cgaaaaagga tggatatacc gaaaaaatcg
     1621 ctataatgac cccgaagcag ggttatgcag cggaaaagcg ctgcttccct gctgttttgt
     1681 ggaatatcta ccgactggaa acaggcaaat gcaggaaatt actgaactga ggggacaggc
     1741 gagagacgat gccaaagagc tacaccgacg agctggccga gtgggttgaa tcccgcgcgg
     1801 ccaagaagcg ccggcgtgat gaggctgcgg ttgcgttcct ggcggtgagg gcggatgtcg
     1861 aggcggcgtt agcgtccggc tatgcgctcg tcaccatttg ggagcacatg cgggaaacgg
     1921 ggaaggtcaa gttctcctac gagacgttcc gctcgcacgc caggcggcac atcaaggcca
     1981 agcccgccga tgtgcccgca ccgcaggcca aggctgcgga acccgcgccg gcacccaaga
     2041 cgccggagcc acggcggccg aagcaggggg gcaaggctga aaagccggcc cccgctgcgg
     2101 ccccgaccgg cttcaccttc aacccaacac cggacaaaaa ggatcctcta cgccggacgc
     2161 atcgtggccg gcatcaccgg cgccacaagt gcggttgctg gcgcctatat cgccgacatc
     2221 accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt
     2281 atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttgct gcctcgcgcg
     2341 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     2401 tctgtaagcg gatgccgtaa gtaaggtaaa ggcctcgcgg aacccctatt tgtttatttt
     2461 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     2521 aatattgaaa aaggagctta tcaccgataa ggaggttttt taatgacggt gctgcactcg
     2581 gttgacttct tccctagcgg caatgccagc gttgccattg agccgcgcct gcctcaagcc
     2641 gacttcccgg agcaccacca cgacttccac gagatcgtta tcgtggagca cggtaccggc
     2701 atccacgttt tcaacggcca accgtacacg attacgggcg gtaccgtgtg cttcgttcgt
     2761 gatcacgacc gccacttata cgagcacacg gacaacttat gcttaaccaa cgttttatac
     2821 cgtagccctg accgcttcca attcctggcg ggcttaaacc aactgttacc gcaggaatta
     2881 gacggccaat accctagcca ttggcgtgtg aatcattcgg tgctgcaaca agttcgccaa
     2941 ttagtggcgc aaatggagca acaagagggc gagaacgacc tgccgagcac ggcgagccgt
     3001 gaaattctgt tcatgcagct gttactgctg ttacgcaagt cgagcctgca agaaaattta
     3061 gagaattcgg cgagccgcct gaatctgctg ttagcgtggt tagaagatca cttcgcggac
     3121 gaagttaact gggatgcggt tgccgaccag ttcagcctga gcttacgcac cctgcaccgc
     3181 caactgaaac aacagaccgg cttaaccccg caacgctatt taaaccgttt acgcttaatg
     3241 aaggcgcgcc acttactgcg ccattcggaa gcgtcggtga ccgatattgc gtaccactgt
     3301 ggcttttcgg atagcaatca tttcagcacc ctgttccgtc gcgaattcaa ttggagccct
     3361 cgcgacatcc gccaaggccg cgacggtttc ttacagtgaa ggccttctag agtcgacctg
     3421 caggcatgca agcttggctg ttttggcgga tgagagggta ccgcatgagc ttcgagctaa
     3481 tcgcgcaaaa aaccccgctt cggcggggtt ttttcgctga tcacgtacga tatcttcgaa
     3541 ccggtgcaca tggtgtacag ggccctagga taggacgtct taaggtttaa accaggttaa
     3601 ttaatttaaa tgcatcccgg gacgtctcga gctcgatcgg accgcggccg ctagcacgta
     3661 taccaagtgt cctgttttct tgccgccaag gatctgatgg cgcaggggat caagatctct
     3721 gttgataccg ggaagccctg ggccaacttt tggcgaaaat gagacgttga tcggcacgta
     3781 agaggttcca actttcacca taatgaaata agatcactac cgggcgtatt ttttgagtta
     3841 tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc
     3901 gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa
     3961 tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa
     4021 aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat
     4081 ccggaattac gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct
     4141 tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac
     4201 gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac
     4261 ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg
     4321 gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt
     4381 ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag
     4441 gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag
     4501 tactgcgatg agtggcaggg cggggcgtaa tttttttaag gcagttattg tcttcaaatt
     4561 cccgttgcat cgatggcccc cgatggtagt gtggggtctc cccatgcgag agtagggaac
     4621 tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
     4681 ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt
     4741 tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca
     4801 aattaagcag aaggccatcc tgacggatgg cctttttgcg tggccagtgc caagcttgca
     4861 tgcctgcagc tgtccggaac tagtcgaggc catggtggcc tcctgatgtc gtcaacacgg
     4921 cgaaatagta atcacgaggt caggttctta ccttaaattt tcgacggaaa accacgtaaa
     4981 aaacgtcgat ttttcaagat acagcgtgaa ttttcaggaa atgcggtgag catcacatca
     5041 ccacaattca gcaaattgtg aacatcatca cgttcatctt tccctggttg ccaatggccc
     5101 attttcctgt cagtaacgag aaggtcgcga attcaggcgc tttttagact ggtcgtaatg
     5161 aaattcagca ggatcaccat atgaggaggt aaaatagatg tcctacgact acgagaaaac
     5221 cagcctcacc ctctaccggg cggtattcaa ggccaactac gacggcgacg tcggtcgcta
     5281 cctgcatccc gacaaggaac tcgccgaggc tgcggaagtc gccccgctgc tgcatccgac
     5341 cttcgacagc cccaacaccc ctggcgtccc cgcccgcgcg ccggacatcg tcgccggccg
     5401 cgacggcctc tacgccccgg acaccggcgg cacctcggtg ttcgaccgcg ccggcgtgct
     5461 gcgccgcgcc gacggcgact tcgtgatacc cgacggcacc gacatcccgc cggaccttaa
     5521 ggtgaagcag gacagctaca acaagcgcct gcaagccacc cactacacca tcatgccggc
     5581 caagccgatg taccgggagg tcctcatggg ccaactggac aacttcgtgc gcaacgccat
     5641 ccgccgccaa tgggaaaaag cccgcgggct ctagggatcc gcgaattctc atgtttgaca
     5701 gcttatcact gatcagtgaa ttaatggcga tgacgcatcc tcacgataat atccgggtag
     5761 gcgcaatcac tttcgtctct actccgttac aaagcgaggc tgggtatttc ccggcctttc
     5821 tgttatccga aatccactga aagcacagcg gctggctgag gagataaata ataaacgagg
     5881 ggctgtatgc acaaagcatc ttctgttgag ttaagaacga gtatcgagat ggcacatagc
     5941 cttgctcaaa ttggaatcag gtttgtgcca ataccagtag aaacagacga agaagtcgtg
     6001 ggggaattcc acaaattgtt atccgctcac aattccacat gtggaattcc acatgtggaa
     6061 ttc
//
