LOCUS       V016130                 6808 bp    DNA     circular SYN 30-JUN-2019
DEFINITION  Exported.
ACCESSION   V016130
VERSION     V016130
KEYWORDS    UNVERIFIED
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6808)
  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 6808)
  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 6808)
  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..6808
                     /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..2220
                     /gene="yhaV"
                     /label="Ribonuclease toxin YhaV"
                     /note="Ribonuclease toxin YhaV from Escherichia coli
                     (strain K12). Accession#: P64594"
     terminator      2283..2529
                     /label="lambda tL3 terminator"
                     /note="transcription terminator tL3 from phage lambda"
     protein_bind    complement(2565..2581)
                     /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(2616..3004)
                     /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(3625..3993)
                     /label="traJ"
                     /note="oriT-recognizing protein"
     oriT            complement(4026..4135)
                     /direction=LEFT
                     /label="oriT"
                     /note="incP origin of transfer"
     regulatory      5048..5065
                     /label="AmpR promoter"
                     /note="AmpR promoter"
                     /regulatory_class="promoter"
     regulatory      5066..5147
                     /label="AmpR promoter"
                     /note="AmpR promoter"
                     /regulatory_class="promoter"
     CDS             5175..6008
                     /label="rhaS"
                     /note="positive regulator of the rhaB promoter"
     promoter        6046..6074
                     /label="tac promoter"
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     CDS             6141..6803
                     /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 ggattttcca caaagggtta atggttgggc gctatatgct
     1801 catccctgtt ttcaggaaac ctacgacgct ttagttgccg aagtcgagac attaaaggga
     1861 aaagatcctg aaaattatca gagaaaagcc gccacaaagt tattggcggt agtccataaa
     1921 gtgattgagg agcatatcac ggtcaatcca tcatcaccgg cattccgtca tggcaagtcg
     1981 ttaggctctg ggaaaaataa agactggtca cgggtaaaat ttggtgctgg tcgttatcgt
     2041 ctcttctttc gttatagtga aaaagagaaa gtcatcattc tgggatggat gaacgatgaa
     2101 aacactctgc gcacctacgg taaaaaaaca gatgcctata ccgtattcag caaaatgtta
     2161 aaaagaggac atcctcctgc cgactgggaa accctcaccc gagaaacaga agaaacccat
     2221 tgaggatccg cgaattctca tgtttgacag cttatcactg atcagtgaat taatggcgat
     2281 gacgcatcct cacgataata tccgggtagg cgcaatcact ttcgtctcta ctccgttaca
     2341 aagcgaggct gggtatttcc cggcctttct gttatccgaa atccactgaa agcacagcgg
     2401 ctggctgagg agataaataa taaacgaggg gctgtatgca caaagcatct tctgttgagt
     2461 taagaacgag tatcgagatg gcacatagcc ttgctcaaat tggaatcagg tttgtgccaa
     2521 taccagtaga aacagacgaa gaagtcgtgg gggaattcca caaattgtta tccgctcaca
     2581 attccacatg tggaattcca catgtggaat tcccatgtca gccgttaagt gttcctgtgt
     2641 cactcaaaat tgctttgaga ggctctaagg gcttctcagt gcgttacatc cctggcttgt
     2701 tgtccacaac cgttaaacct taaaagcttt aaaagcctta tatattcttt tttttcttat
     2761 aaaacttaaa accttagagg ctatttaagt tgctgattta tattaatttt attgttcaaa
     2821 catgagagct tagtacgtga aacatgagag cttagtacgt tagccatgag agcttagtac
     2881 gttagccatg agggtttagt tcgttaaaca tgagagctta gtacgttaaa catgagagct
     2941 tagtacgtga aacatgagag cttagtacgt actatcaaca ggttgaactg ctgatcttca
     3001 gatcctctac gccggacgca tcgtggccgg atccagccga ccaggctttc cacgcccgcg
     3061 tgccgctcca tgtcgttcgc gcggttctcg gaaacgcgct gccgcgtttc gtgattgtca
     3121 cgctcaagcc cgtagtcccg ttcgagcgtc gcgcagaggt cagcgagggc gcggtaggcc
     3181 cgatacggct catggatggt gtttcgggtc gggtgaatct tgttgatggc gatatggatg
     3241 tgcaggttgt cggtgtcgtg atgcacggca ctgacgcgct gatgctcggc gaagccaagc
     3301 ccagcgcaga tgcggtcctc aatcgcgcgc aacgtctccg cgtcgggctt ctctcccgcg
     3361 cggaagctaa ccagcaggtg ataggtcttg tcggcctcgg aacgggtgtt gccgtgctgg
     3421 gtcgccatca cctcggccat gacagcgggc agggtgtttg cctcgcagtt cgtgacgcgc
     3481 acgtgaccca ggcgctcggt cttgccttgc tcgtcggtga tgtacttcac cagctccgcg
     3541 aagtcgctct tcttgatgga gcgcatgggg acgtgcttgg caatcacgcg caccccccgg
     3601 ccgttttagc ggctaaaaaa gtcatggctc tgccctcggg cggaccacgc ccatcatgac
     3661 cttgccaagc tcgtcctgct tctcttcgat cttcgccagc agggcgagga tcgtggcatc
     3721 accgaaccgc gccgtgcgcg ggtcgtcggt gagccagagt ttcagcaggc cgcccaggcg
     3781 gcccaggtcg ccattgatgc gggccagctc gcggacgtgc tcatagtcca cgacgcccgt
     3841 gattttgtag ccctggccga cggccagcag gtaggccgac aggctcatgc cggccgccgc
     3901 cgccttttcc tcaatcgctc ttcgttcgtc tggaaggcag tacaccttga taggtgggct
     3961 gcccttcctg gttggcttgg tttcatcagc catccgcttg ccctcatctg ttacgccggc
     4021 ggtagccggc cagcctcgca gagcaggatt cccgttgagc accgccaggt gcgaataagg
     4081 gacagtgaag aaggaacacc cgctcgcggg tgggcctact tcacctatcc tgcccggctg
     4141 acgccgttgg atacaccaag gaaagtctac acgaaccctt tggcaaaatc ctgtatatcg
     4201 tgcgaaaaag gatggatata ccgaaaaaat cgctataatg accccgaagc agggttatgc
     4261 agcggaaaag cgctgcttcc ctgctgtttt gtggaatatc taccgactgg aaacaggcaa
     4321 atgcaggaaa ttactgaact gaggggacag gcgagagacg atgccaaaga gctacaccga
     4381 cgagctggcc gagtgggttg aatcccgcgc ggccaagaag cgccggcgtg atgaggctgc
     4441 ggttgcgttc ctggcggtga gggcggatgt cgaggcggcg ttagcgtccg gctatgcgct
     4501 cgtcaccatt tgggagcaca tgcgggaaac ggggaaggtc aagttctcct acgagacgtt
     4561 ccgctcgcac gccaggcggc acatcaaggc caagcccgcc gatgtgcccg caccgcaggc
     4621 caaggctgcg gaacccgcgc cggcacccaa gacgccggag ccacggcggc cgaagcaggg
     4681 gggcaaggct gaaaagccgg cccccgctgc ggccccgacc ggcttcacct tcaacccaac
     4741 accggacaaa aaggatcctc tacgccggac gcatcgtggc cggcatcacc ggcgccacaa
     4801 gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg gctcgccact
     4861 tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg gccgggggac
     4921 tgttgggcgc catctccttg ctgcctcgcg cgtttcggtg atgacggtga aaacctctga
     4981 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgt aagtaaggta
     5041 aaggcctcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     5101 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggagct tatcaccgat
     5161 aaggaggttt tttaatgacg gtgctgcact cggttgactt cttccctagc ggcaatgcca
     5221 gcgttgccat tgagccgcgc ctgcctcaag ccgacttccc ggagcaccac cacgacttcc
     5281 acgagatcgt tatcgtggag cacggtaccg gcatccacgt tttcaacggc caaccgtaca
     5341 cgattacggg cggtaccgtg tgcttcgttc gtgatcacga ccgccactta tacgagcaca
     5401 cggacaactt atgcttaacc aacgttttat accgtagccc tgaccgcttc caattcctgg
     5461 cgggcttaaa ccaactgtta ccgcaggaat tagacggcca ataccctagc cattggcgtg
     5521 tgaatcattc ggtgctgcaa caagttcgcc aattagtggc gcaaatggag caacaagagg
     5581 gcgagaacga cctgccgagc acggcgagcc gtgaaattct gttcatgcag ctgttactgc
     5641 tgttacgcaa gtcgagcctg caagaaaatt tagagaattc ggcgagccgc ctgaatctgc
     5701 tgttagcgtg gttagaagat cacttcgcgg acgaagttaa ctgggatgcg gttgccgacc
     5761 agttcagcct gagcttacgc accctgcacc gccaactgaa acaacagacc ggcttaaccc
     5821 cgcaacgcta tttaaaccgt ttacgcttaa tgaaggcgcg ccacttactg cgccattcgg
     5881 aagcgtcggt gaccgatatt gcgtaccact gtggcttttc ggatagcaat catttcagca
     5941 ccctgttccg tcgcgaattc aattggagcc ctcgcgacat ccgccaaggc cgcgacggtt
     6001 tcttacagtg aaggccttct agagtcgacc tgcatcgacc tgcagttgac aattaatcat
     6061 cggctcgtat aatgtgtgga aaggcggttc accgccgttt tcacacagga aacagaattc
     6121 tttaagaagg agatatacat atgtcagtga tagcaaagca gatgacatac aaagtatata
     6181 tgagcggtac tgtaaatggt cactacttcg aagtagaagg tgatggcaaa gggaagccgt
     6241 atgagggtga acaaacagtg aagcttacag ttacgaaggg tggccctttg ccgttcgcgt
     6301 gggacatttt gtcgccacag tgccagtacg ggagcatacc attcaccaag tatccagagg
     6361 acataccaga ctacgtgaaa cagtcctttc ccgaaggcta tacctgggag cgcataatga
     6421 actttgaaga cggtgcggtt tgtacggtat cgaacgattc atcaatccaa ggaaattgct
     6481 ttatttatca tgttaaattt tcggggctta actttccgcc aaatggcccc gtgatgcaga
     6541 agaaaactca gggctgggaa ccaaataccg agcgcctttt cgctcgggac ggaatgctgt
     6601 tgggaaacaa ttttatggcg ttgaagctgg aagggggtgg ccactatctc tgtgaattca
     6661 agacaacata caaagccaag aagcctgtca agatgcccgg ttatcactat gtggaccgga
     6721 agctcgatgt cactaatcac aacaaggatt atacttcagt tgaacagtgc gaaatttcga
     6781 tcgctcgcaa acctgtcgta gcataagg
//
