LOCUS       V015634                 6826 bp    DNA     circular SYN 30-JUN-2019
DEFINITION  Exported.
ACCESSION   V015634
VERSION     V015634
KEYWORDS    UNVERIFIED
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6826)
  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 6826)
  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 6826)
  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..6826
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      42..75
                     /label="strong forward terminator"
                     /note="strong forward terminator"
                     /regulatory_class="terminator"
     promoter        326..428
                     /label="cat promoter"
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             429..1085
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     terminator      1189..1275
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      1367..1394
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        1597..1715
                     /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             1756..2229
                     /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      2292..2538
                     /label="lambda tL3 terminator"
                     /note="transcription terminator tL3 from phage lambda"
     protein_bind    complement(2574..2590)
                     /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(2625..3013)
                     /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(3634..4002)
                     /label="traJ"
                     /note="oriT-recognizing protein"
     oriT            complement(4035..4144)
                     /direction=LEFT
                     /label="oriT"
                     /note="incP origin of transfer"
     regulatory      5057..5074
                     /label="AmpR promoter"
                     /note="AmpR promoter"
                     /regulatory_class="promoter"
     regulatory      5075..5156
                     /label="AmpR promoter"
                     /note="AmpR promoter"
                     /regulatory_class="promoter"
     CDS             5184..6017
                     /label="rhaS"
                     /note="positive regulator of the rhaB promoter"
     gene            6044..6090
                     /gene="apFAB46"
                     /label="apFAB46"
     misc_feature    6044..6090
                     /gene="apFAB46"
                     /label="similar to apFAB46"
                     /note="similar to apFAB46"
     gene            6099..6110
                     /gene="B0034"
                     /label="B0034"
     misc_feature    6099..6110
                     /gene="B0034"
                     /label="similar to B0034"
                     /note="similar to B0034"
     CDS             6117..6800
                     /label="tsPurple"
                     /note="tsPurple is a basic (constitutively fluorescent)
                     fluorescent protein published in 2018, derived from
                     synthetic construct."
ORIGIN
        1 ttggcggatg agagggtacc gcatgagctt cgagctaatc gcgcaaaaaa ccccgcttcg
       61 gcggggtttt ttcgctgatc acgtacgata tcttcgaacc ggtgcacatg gtgtacaggg
      121 ccctaggata ggacgtctta aggtttaaac caggttaatt aatttaaatg catcccggga
      181 cgtctcgagc tcgatcggac cgcggccgct agcacgtata ccaagtgtcc tgttttcttg
      241 ccgccaagga tctgatggcg caggggatca agatctctgt tgataccggg aagccctggg
      301 ccaacttttg gcgaaaatga gacgttgatc ggcacgtaag aggttccaac tttcaccata
      361 atgaaataag atcactaccg ggcgtatttt ttgagttatc gagattttca ggagctaagg
      421 aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc
      481 gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc
      541 agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg
      601 cctttattca cattcttgcc cgcctgatga atgctcatcc ggaattacgt atggcaatga
      661 aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc
      721 aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac
      781 acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt
      841 ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt
      901 taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata
      961 cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg
     1021 gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg
     1081 gggcgtaatt tttttaaggc agttattgtc ttcaaattcc cgttgcatcg atggcccccg
     1141 atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga
     1201 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     1261 ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg
     1321 tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg
     1381 acggatggcc tttttgcgtg gccagtgcca agcttgcatg cctgcagctg tccggaacta
     1441 gtcgaggcca tggtggcctc ctgatgtcgt caacacggcg aaatagtaat cacgaggtca
     1501 ggttcttacc ttaaattttc gacggaaaac cacgtaaaaa acgtcgattt ttcaagatac
     1561 agcgtgaatt ttcaggaaat gcggtgagca tcacatcacc acaattcagc aaattgtgaa
     1621 catcatcacg ttcatctttc cctggttgcc aatggcccat tttcctgtca gtaacgagaa
     1681 ggtcgcgaat tcaggcgctt tttagactgg tcgtaatgaa attcagcagg atcaccatat
     1741 gaggaggtaa aatagatgtc ctacgactac gagaaaacca gcctcaccct ctaccgggcg
     1801 gtattcaagg ccaactacga cggcgacgtc ggtcgctacc tgcatcccga caaggaactc
     1861 gccgaggctg cggaagtcgc cccgctgctg catccgacct tcgacagccc caacacccct
     1921 ggcgtccccg cccgcgcgcc ggacatcgtc gccggccgcg acggcctcta cgccccggac
     1981 accggcggca cctcggtgtt cgaccgcgcc ggcgtgctgc gccgcgccga cggcgacttc
     2041 gtgatacccg acggcaccga catcccgccg gaccttaagg tgaagcagga cagctacaac
     2101 aagcgcctgc aagccaccca ctacaccatc atgccggcca agccgatgta ccgggaggtc
     2161 ctcatgggcc aactggacaa cttcgtgcgc aacgccatcc gccgccaatg ggaaaaagcc
     2221 cgcgggctct agggatccgc gaattctcat gtttgacagc ttatcactga tcagtgaatt
     2281 aatggcgatg acgcatcctc acgataatat ccgggtaggc gcaatcactt tcgtctctac
     2341 tccgttacaa agcgaggctg ggtatttccc ggcctttctg ttatccgaaa tccactgaaa
     2401 gcacagcggc tggctgagga gataaataat aaacgagggg ctgtatgcac aaagcatctt
     2461 ctgttgagtt aagaacgagt atcgagatgg cacatagcct tgctcaaatt ggaatcaggt
     2521 ttgtgccaat accagtagaa acagacgaag aagtcgtggg ggaattccac aaattgttat
     2581 ccgctcacaa ttccacatgt ggaattccac atgtggaatt cccatgtcag ccgttaagtg
     2641 ttcctgtgtc actcaaaatt gctttgagag gctctaaggg cttctcagtg cgttacatcc
     2701 ctggcttgtt gtccacaacc gttaaacctt aaaagcttta aaagccttat atattctttt
     2761 ttttcttata aaacttaaaa ccttagaggc tatttaagtt gctgatttat attaatttta
     2821 ttgttcaaac atgagagctt agtacgtgaa acatgagagc ttagtacgtt agccatgaga
     2881 gcttagtacg ttagccatga gggtttagtt cgttaaacat gagagcttag tacgttaaac
     2941 atgagagctt agtacgtgaa acatgagagc ttagtacgta ctatcaacag gttgaactgc
     3001 tgatcttcag atcctctacg ccggacgcat cgtggccgga tccagccgac caggctttcc
     3061 acgcccgcgt gccgctccat gtcgttcgcg cggttctcgg aaacgcgctg ccgcgtttcg
     3121 tgattgtcac gctcaagccc gtagtcccgt tcgagcgtcg cgcagaggtc agcgagggcg
     3181 cggtaggccc gatacggctc atggatggtg tttcgggtcg ggtgaatctt gttgatggcg
     3241 atatggatgt gcaggttgtc ggtgtcgtga tgcacggcac tgacgcgctg atgctcggcg
     3301 aagccaagcc cagcgcagat gcggtcctca atcgcgcgca acgtctccgc gtcgggcttc
     3361 tctcccgcgc ggaagctaac cagcaggtga taggtcttgt cggcctcgga acgggtgttg
     3421 ccgtgctggg tcgccatcac ctcggccatg acagcgggca gggtgtttgc ctcgcagttc
     3481 gtgacgcgca cgtgacccag gcgctcggtc ttgccttgct cgtcggtgat gtacttcacc
     3541 agctccgcga agtcgctctt cttgatggag cgcatgggga cgtgcttggc aatcacgcgc
     3601 accccccggc cgttttagcg gctaaaaaag tcatggctct gccctcgggc ggaccacgcc
     3661 catcatgacc ttgccaagct cgtcctgctt ctcttcgatc ttcgccagca gggcgaggat
     3721 cgtggcatca ccgaaccgcg ccgtgcgcgg gtcgtcggtg agccagagtt tcagcaggcc
     3781 gcccaggcgg cccaggtcgc cattgatgcg ggccagctcg cggacgtgct catagtccac
     3841 gacgcccgtg attttgtagc cctggccgac ggccagcagg taggccgaca ggctcatgcc
     3901 ggccgccgcc gccttttcct caatcgctct tcgttcgtct ggaaggcagt acaccttgat
     3961 aggtgggctg cccttcctgg ttggcttggt ttcatcagcc atccgcttgc cctcatctgt
     4021 tacgccggcg gtagccggcc agcctcgcag agcaggattc ccgttgagca ccgccaggtg
     4081 cgaataaggg acagtgaaga aggaacaccc gctcgcgggt gggcctactt cacctatcct
     4141 gcccggctga cgccgttgga tacaccaagg aaagtctaca cgaacccttt ggcaaaatcc
     4201 tgtatatcgt gcgaaaaagg atggatatac cgaaaaaatc gctataatga ccccgaagca
     4261 gggttatgca gcggaaaagc gctgcttccc tgctgttttg tggaatatct accgactgga
     4321 aacaggcaaa tgcaggaaat tactgaactg aggggacagg cgagagacga tgccaaagag
     4381 ctacaccgac gagctggccg agtgggttga atcccgcgcg gccaagaagc gccggcgtga
     4441 tgaggctgcg gttgcgttcc tggcggtgag ggcggatgtc gaggcggcgt tagcgtccgg
     4501 ctatgcgctc gtcaccattt gggagcacat gcgggaaacg gggaaggtca agttctccta
     4561 cgagacgttc cgctcgcacg ccaggcggca catcaaggcc aagcccgccg atgtgcccgc
     4621 accgcaggcc aaggctgcgg aacccgcgcc ggcacccaag acgccggagc cacggcggcc
     4681 gaagcagggg ggcaaggctg aaaagccggc ccccgctgcg gccccgaccg gcttcacctt
     4741 caacccaaca ccggacaaaa aggatcctct acgccggacg catcgtggcc ggcatcaccg
     4801 gcgccacaag tgcggttgct ggcgcctata tcgccgacat caccgatggg gaagatcggg
     4861 ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg
     4921 ccgggggact gttgggcgcc atctccttgc tgcctcgcgc gtttcggtga tgacggtgaa
     4981 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccgta
     5041 agtaaggtaa aggcctcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     5101 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggagctt
     5161 atcaccgata aggaggtttt ttaatgacgg tgctgcactc ggttgacttc ttccctagcg
     5221 gcaatgccag cgttgccatt gagccgcgcc tgcctcaagc cgacttcccg gagcaccacc
     5281 acgacttcca cgagatcgtt atcgtggagc acggtaccgg catccacgtt ttcaacggcc
     5341 aaccgtacac gattacgggc ggtaccgtgt gcttcgttcg tgatcacgac cgccacttat
     5401 acgagcacac ggacaactta tgcttaacca acgttttata ccgtagccct gaccgcttcc
     5461 aattcctggc gggcttaaac caactgttac cgcaggaatt agacggccaa taccctagcc
     5521 attggcgtgt gaatcattcg gtgctgcaac aagttcgcca attagtggcg caaatggagc
     5581 aacaagaggg cgagaacgac ctgccgagca cggcgagccg tgaaattctg ttcatgcagc
     5641 tgttactgct gttacgcaag tcgagcctgc aagaaaattt agagaattcg gcgagccgcc
     5701 tgaatctgct gttagcgtgg ttagaagatc acttcgcgga cgaagttaac tgggatgcgg
     5761 ttgccgacca gttcagcctg agcttacgca ccctgcaccg ccaactgaaa caacagaccg
     5821 gcttaacccc gcaacgctat ttaaaccgtt tacgcttaat gaaggcgcgc cacttactgc
     5881 gccattcgga agcgtcggtg accgatattg cgtaccactg tggcttttcg gatagcaatc
     5941 atttcagcac cctgttccgt cgcgaattca attggagccc tcgcgacatc cgccaaggcc
     6001 gcgacggttt cttacagtga aggccttcta gagtcgacct gcaaaaaaga gtattgactt
     6061 cgcatctttt tgtacctata atagattcat tactagagaa agaggagaaa tactagatgg
     6121 cgtcccttgt aaagaaggat atgtgtgtta agatgacaat ggagggaacc gtcaacgggt
     6181 atcactttaa gtgtgttggc gagggagaag gcaaaccgtt cgagggtaca cagaatatgc
     6241 gcattcgtgt caccgaaggg ggccctttgc cctttgcttt cgatattttg gccccgtgct
     6301 gtatgtatgg ttcgaagact tttatcaaac acgtaagcgg aatccctgac tacttcaagg
     6361 agagctttcc cgaaggcttt acttgggaac gtacgcagat ttttgaagac ggaggagttt
     6421 tgactgccca tcaagataca tcacttgagg gaaactgttt aatctacaaa gttaaggtgt
     6481 tagggacaaa ttttcctgcc aatggtccgg ttatgcagaa aaagacagca ggctgggaac
     6541 cgtgtgtaga gatgttgtac ccccgcgacg gggttctttg cgggcaatcc cttatggcct
     6601 tgaaatgtac tgatggaaat cacttgacat cccatttacg caccacttat cgctcgcgta
     6661 agccaagtaa tgcggtgaac atgccggaat ttcacttcgg agaccatcgt attgaaatct
     6721 tgaaagccga acaaggaaaa ttttatgaac aatacgaatc cgcagtggca cgttattcag
     6781 atgttccaga aaaggcgact tgataaggca tgcaagcttg gctgtt
//
