LOCUS       V016133                 6797 bp    DNA     circular SYN 30-JUN-2019
DEFINITION  Exported.
ACCESSION   V016133
VERSION     V016133
KEYWORDS    UNVERIFIED
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6797)
  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 6797)
  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 6797)
  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..6797
                     /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             1739..2200
                     /gene="yhaV"
                     /label="Ribonuclease toxin YhaV"
                     /note="Ribonuclease toxin YhaV from Escherichia coli
                     (strain K12). Accession#: P64594"
     terminator      2263..2509
                     /label="lambda tL3 terminator"
                     /note="transcription terminator tL3 from phage lambda"
     protein_bind    complement(2545..2561)
                     /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(2596..2984)
                     /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(3605..3973)
                     /label="traJ"
                     /note="oriT-recognizing protein"
     oriT            complement(4006..4115)
                     /direction=LEFT
                     /label="oriT"
                     /note="incP origin of transfer"
     regulatory      5028..5045
                     /label="AmpR promoter"
                     /note="AmpR promoter"
                     /regulatory_class="promoter"
     regulatory      5046..5127
                     /label="AmpR promoter"
                     /note="AmpR promoter"
                     /regulatory_class="promoter"
     CDS             5155..5988
                     /label="rhaS"
                     /note="positive regulator of the rhaB promoter"
     gene            6015..6061
                     /gene="apFAB46"
                     /label="apFAB46"
     misc_feature    6015..6061
                     /gene="apFAB46"
                     /label="similar to apFAB46"
                     /note="similar to apFAB46"
     gene            6070..6081
                     /gene="B0034"
                     /label="B0034"
     misc_feature    6070..6081
                     /gene="B0034"
                     /label="similar to B0034"
                     /note="similar to B0034"
     CDS             6088..6771
                     /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 ggattttcca caaagggtta atggttgggc gctatatgct catccctgtt ttcaggaaac
     1801 ctacgacgct ttagttgccg aagtcgagac attaaaggga aaagatcctg aaaattatca
     1861 gagaaaagcc gccacaaagt tattggcggt agtccataaa gtgattgagg agcatatcac
     1921 ggtcaatcca tcatcaccgg cattccgtca tggcaagtcg ttaggctctg ggaaaaataa
     1981 agactggtca cgggtaaaat ttggtgctgg tcgttatcgt ctcttctttc gttatagtga
     2041 aaaagagaaa gtcatcattc tgggatggat gaacgatgaa aacactctgc gcacctacgg
     2101 taaaaaaaca gatgcctata ccgtattcag caaaatgtta aaaagaggac atcctcctgc
     2161 cgactgggaa accctcaccc gagaaacaga agaaacccat tgaggatccg cgaattctca
     2221 tgtttgacag cttatcactg atcagtgaat taatggcgat gacgcatcct cacgataata
     2281 tccgggtagg cgcaatcact ttcgtctcta ctccgttaca aagcgaggct gggtatttcc
     2341 cggcctttct gttatccgaa atccactgaa agcacagcgg ctggctgagg agataaataa
     2401 taaacgaggg gctgtatgca caaagcatct tctgttgagt taagaacgag tatcgagatg
     2461 gcacatagcc ttgctcaaat tggaatcagg tttgtgccaa taccagtaga aacagacgaa
     2521 gaagtcgtgg gggaattcca caaattgtta tccgctcaca attccacatg tggaattcca
     2581 catgtggaat tcccatgtca gccgttaagt gttcctgtgt cactcaaaat tgctttgaga
     2641 ggctctaagg gcttctcagt gcgttacatc cctggcttgt tgtccacaac cgttaaacct
     2701 taaaagcttt aaaagcctta tatattcttt tttttcttat aaaacttaaa accttagagg
     2761 ctatttaagt tgctgattta tattaatttt attgttcaaa catgagagct tagtacgtga
     2821 aacatgagag cttagtacgt tagccatgag agcttagtac gttagccatg agggtttagt
     2881 tcgttaaaca tgagagctta gtacgttaaa catgagagct tagtacgtga aacatgagag
     2941 cttagtacgt actatcaaca ggttgaactg ctgatcttca gatcctctac gccggacgca
     3001 tcgtggccgg atccagccga ccaggctttc cacgcccgcg tgccgctcca tgtcgttcgc
     3061 gcggttctcg gaaacgcgct gccgcgtttc gtgattgtca cgctcaagcc cgtagtcccg
     3121 ttcgagcgtc gcgcagaggt cagcgagggc gcggtaggcc cgatacggct catggatggt
     3181 gtttcgggtc gggtgaatct tgttgatggc gatatggatg tgcaggttgt cggtgtcgtg
     3241 atgcacggca ctgacgcgct gatgctcggc gaagccaagc ccagcgcaga tgcggtcctc
     3301 aatcgcgcgc aacgtctccg cgtcgggctt ctctcccgcg cggaagctaa ccagcaggtg
     3361 ataggtcttg tcggcctcgg aacgggtgtt gccgtgctgg gtcgccatca cctcggccat
     3421 gacagcgggc agggtgtttg cctcgcagtt cgtgacgcgc acgtgaccca ggcgctcggt
     3481 cttgccttgc tcgtcggtga tgtacttcac cagctccgcg aagtcgctct tcttgatgga
     3541 gcgcatgggg acgtgcttgg caatcacgcg caccccccgg ccgttttagc ggctaaaaaa
     3601 gtcatggctc tgccctcggg cggaccacgc ccatcatgac cttgccaagc tcgtcctgct
     3661 tctcttcgat cttcgccagc agggcgagga tcgtggcatc accgaaccgc gccgtgcgcg
     3721 ggtcgtcggt gagccagagt ttcagcaggc cgcccaggcg gcccaggtcg ccattgatgc
     3781 gggccagctc gcggacgtgc tcatagtcca cgacgcccgt gattttgtag ccctggccga
     3841 cggccagcag gtaggccgac aggctcatgc cggccgccgc cgccttttcc tcaatcgctc
     3901 ttcgttcgtc tggaaggcag tacaccttga taggtgggct gcccttcctg gttggcttgg
     3961 tttcatcagc catccgcttg ccctcatctg ttacgccggc ggtagccggc cagcctcgca
     4021 gagcaggatt cccgttgagc accgccaggt gcgaataagg gacagtgaag aaggaacacc
     4081 cgctcgcggg tgggcctact tcacctatcc tgcccggctg acgccgttgg atacaccaag
     4141 gaaagtctac acgaaccctt tggcaaaatc ctgtatatcg tgcgaaaaag gatggatata
     4201 ccgaaaaaat cgctataatg accccgaagc agggttatgc agcggaaaag cgctgcttcc
     4261 ctgctgtttt gtggaatatc taccgactgg aaacaggcaa atgcaggaaa ttactgaact
     4321 gaggggacag gcgagagacg atgccaaaga gctacaccga cgagctggcc gagtgggttg
     4381 aatcccgcgc ggccaagaag cgccggcgtg atgaggctgc ggttgcgttc ctggcggtga
     4441 gggcggatgt cgaggcggcg ttagcgtccg gctatgcgct cgtcaccatt tgggagcaca
     4501 tgcgggaaac ggggaaggtc aagttctcct acgagacgtt ccgctcgcac gccaggcggc
     4561 acatcaaggc caagcccgcc gatgtgcccg caccgcaggc caaggctgcg gaacccgcgc
     4621 cggcacccaa gacgccggag ccacggcggc cgaagcaggg gggcaaggct gaaaagccgg
     4681 cccccgctgc ggccccgacc ggcttcacct tcaacccaac accggacaaa aaggatcctc
     4741 tacgccggac gcatcgtggc cggcatcacc ggcgccacaa gtgcggttgc tggcgcctat
     4801 atcgccgaca tcaccgatgg ggaagatcgg gctcgccact tcgggctcat gagcgcttgt
     4861 ttcggcgtgg gtatggtggc aggccccgtg gccgggggac tgttgggcgc catctccttg
     4921 ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     4981 ggtcacagct tgtctgtaag cggatgccgt aagtaaggta aaggcctcgc ggaaccccta
     5041 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     5101 aaatgcttca ataatattga aaaaggagct tatcaccgat aaggaggttt tttaatgacg
     5161 gtgctgcact cggttgactt cttccctagc ggcaatgcca gcgttgccat tgagccgcgc
     5221 ctgcctcaag ccgacttccc ggagcaccac cacgacttcc acgagatcgt tatcgtggag
     5281 cacggtaccg gcatccacgt tttcaacggc caaccgtaca cgattacggg cggtaccgtg
     5341 tgcttcgttc gtgatcacga ccgccactta tacgagcaca cggacaactt atgcttaacc
     5401 aacgttttat accgtagccc tgaccgcttc caattcctgg cgggcttaaa ccaactgtta
     5461 ccgcaggaat tagacggcca ataccctagc cattggcgtg tgaatcattc ggtgctgcaa
     5521 caagttcgcc aattagtggc gcaaatggag caacaagagg gcgagaacga cctgccgagc
     5581 acggcgagcc gtgaaattct gttcatgcag ctgttactgc tgttacgcaa gtcgagcctg
     5641 caagaaaatt tagagaattc ggcgagccgc ctgaatctgc tgttagcgtg gttagaagat
     5701 cacttcgcgg acgaagttaa ctgggatgcg gttgccgacc agttcagcct gagcttacgc
     5761 accctgcacc gccaactgaa acaacagacc ggcttaaccc cgcaacgcta tttaaaccgt
     5821 ttacgcttaa tgaaggcgcg ccacttactg cgccattcgg aagcgtcggt gaccgatatt
     5881 gcgtaccact gtggcttttc ggatagcaat catttcagca ccctgttccg tcgcgaattc
     5941 aattggagcc ctcgcgacat ccgccaaggc cgcgacggtt tcttacagtg aaggccttct
     6001 agagtcgacc tgcaaaaaag agtattgact tcgcatcttt ttgtacctat aatagattca
     6061 ttactagaga aagaggagaa atactagatg gcgtcccttg taaagaagga tatgtgtgtt
     6121 aagatgacaa tggagggaac cgtcaacggg tatcacttta agtgtgttgg cgagggagaa
     6181 ggcaaaccgt tcgagggtac acagaatatg cgcattcgtg tcaccgaagg gggccctttg
     6241 ccctttgctt tcgatatttt ggccccgtgc tgtatgtatg gttcgaagac ttttatcaaa
     6301 cacgtaagcg gaatccctga ctacttcaag gagagctttc ccgaaggctt tacttgggaa
     6361 cgtacgcaga tttttgaaga cggaggagtt ttgactgccc atcaagatac atcacttgag
     6421 ggaaactgtt taatctacaa agttaaggtg ttagggacaa attttcctgc caatggtccg
     6481 gttatgcaga aaaagacagc aggctgggaa ccgtgtgtag agatgttgta cccccgcgac
     6541 ggggttcttt gcgggcaatc ccttatggcc ttgaaatgta ctgatggaaa tcacttgaca
     6601 tcccatttac gcaccactta tcgctcgcgt aagccaagta atgcggtgaa catgccggaa
     6661 tttcacttcg gagaccatcg tattgaaatc ttgaaagccg aacaaggaaa attttatgaa
     6721 caatacgaat ccgcagtggc acgttattca gatgttccag aaaaggcgac ttgataaggc
     6781 atgcaagctt ggctgtt
//
