LOCUS       V016408                 6837 bp    DNA     circular SYN 30-JUN-2019
DEFINITION  Exported.
ACCESSION   V016408
VERSION     V016408
KEYWORDS    UNVERIFIED
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6837)
  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 6837)
  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 6837)
  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..6837
                     /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             1776..2249
                     /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      2312..2558
                     /label="lambda tL3 terminator"
                     /note="transcription terminator tL3 from phage lambda"
     protein_bind    complement(2594..2610)
                     /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(2645..3033)
                     /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(3654..4022)
                     /label="traJ"
                     /note="oriT-recognizing protein"
     oriT            complement(4055..4164)
                     /direction=LEFT
                     /label="oriT"
                     /note="incP origin of transfer"
     regulatory      5077..5094
                     /label="AmpR promoter"
                     /note="AmpR promoter"
                     /regulatory_class="promoter"
     regulatory      5095..5176
                     /label="AmpR promoter"
                     /note="AmpR promoter"
                     /regulatory_class="promoter"
     CDS             5204..6037
                     /label="rhaS"
                     /note="positive regulator of the rhaB promoter"
     promoter        6075..6103
                     /label="tac promoter"
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     CDS             6170..6832
                     /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 gaggaggtaa aatagatgtc ctacgactac gagaaaacca
     1801 gcctcaccct ctaccgggcg gtattcaagg ccaactacga cggcgacgtc ggtcgctacc
     1861 tgcatcccga caaggaactc gccgaggctg cggaagtcgc cccgctgctg catccgacct
     1921 tcgacagccc caacacccct ggcgtccccg cccgcgcgcc ggacatcgtc gccggccgcg
     1981 acggcctcta cgccccggac accggcggca cctcggtgtt cgaccgcgcc ggcgtgctgc
     2041 gccgcgccga cggcgacttc gtgatacccg acggcaccga catcccgccg gaccttaagg
     2101 tgaagcagga cagctacaac aagcgcctgc aagccaccca ctacaccatc atgccggcca
     2161 agccgatgta ccgggaggtc ctcatgggcc aactggacaa cttcgtgcgc aacgccatcc
     2221 gccgccaatg ggaaaaagcc cgcgggctct agggatccgc gaattctcat gtttgacagc
     2281 ttatcactga tcagtgaatt aatggcgatg acgcatcctc acgataatat ccgggtaggc
     2341 gcaatcactt tcgtctctac tccgttacaa agcgaggctg ggtatttccc ggcctttctg
     2401 ttatccgaaa tccactgaaa gcacagcggc tggctgagga gataaataat aaacgagggg
     2461 ctgtatgcac aaagcatctt ctgttgagtt aagaacgagt atcgagatgg cacatagcct
     2521 tgctcaaatt ggaatcaggt ttgtgccaat accagtagaa acagacgaag aagtcgtggg
     2581 ggaattccac aaattgttat ccgctcacaa ttccacatgt ggaattccac atgtggaatt
     2641 cccatgtcag ccgttaagtg ttcctgtgtc actcaaaatt gctttgagag gctctaaggg
     2701 cttctcagtg cgttacatcc ctggcttgtt gtccacaacc gttaaacctt aaaagcttta
     2761 aaagccttat atattctttt ttttcttata aaacttaaaa ccttagaggc tatttaagtt
     2821 gctgatttat attaatttta ttgttcaaac atgagagctt agtacgtgaa acatgagagc
     2881 ttagtacgtt agccatgaga gcttagtacg ttagccatga gggtttagtt cgttaaacat
     2941 gagagcttag tacgttaaac atgagagctt agtacgtgaa acatgagagc ttagtacgta
     3001 ctatcaacag gttgaactgc tgatcttcag atcctctacg ccggacgcat cgtggccgga
     3061 tccagccgac caggctttcc acgcccgcgt gccgctccat gtcgttcgcg cggttctcgg
     3121 aaacgcgctg ccgcgtttcg tgattgtcac gctcaagccc gtagtcccgt tcgagcgtcg
     3181 cgcagaggtc agcgagggcg cggtaggccc gatacggctc atggatggtg tttcgggtcg
     3241 ggtgaatctt gttgatggcg atatggatgt gcaggttgtc ggtgtcgtga tgcacggcac
     3301 tgacgcgctg atgctcggcg aagccaagcc cagcgcagat gcggtcctca atcgcgcgca
     3361 acgtctccgc gtcgggcttc tctcccgcgc ggaagctaac cagcaggtga taggtcttgt
     3421 cggcctcgga acgggtgttg ccgtgctggg tcgccatcac ctcggccatg acagcgggca
     3481 gggtgtttgc ctcgcagttc gtgacgcgca cgtgacccag gcgctcggtc ttgccttgct
     3541 cgtcggtgat gtacttcacc agctccgcga agtcgctctt cttgatggag cgcatgggga
     3601 cgtgcttggc aatcacgcgc accccccggc cgttttagcg gctaaaaaag tcatggctct
     3661 gccctcgggc ggaccacgcc catcatgacc ttgccaagct cgtcctgctt ctcttcgatc
     3721 ttcgccagca gggcgaggat cgtggcatca ccgaaccgcg ccgtgcgcgg gtcgtcggtg
     3781 agccagagtt tcagcaggcc gcccaggcgg cccaggtcgc cattgatgcg ggccagctcg
     3841 cggacgtgct catagtccac gacgcccgtg attttgtagc cctggccgac ggccagcagg
     3901 taggccgaca ggctcatgcc ggccgccgcc gccttttcct caatcgctct tcgttcgtct
     3961 ggaaggcagt acaccttgat aggtgggctg cccttcctgg ttggcttggt ttcatcagcc
     4021 atccgcttgc cctcatctgt tacgccggcg gtagccggcc agcctcgcag agcaggattc
     4081 ccgttgagca ccgccaggtg cgaataaggg acagtgaaga aggaacaccc gctcgcgggt
     4141 gggcctactt cacctatcct gcccggctga cgccgttgga tacaccaagg aaagtctaca
     4201 cgaacccttt ggcaaaatcc tgtatatcgt gcgaaaaagg atggatatac cgaaaaaatc
     4261 gctataatga ccccgaagca gggttatgca gcggaaaagc gctgcttccc tgctgttttg
     4321 tggaatatct accgactgga aacaggcaaa tgcaggaaat tactgaactg aggggacagg
     4381 cgagagacga tgccaaagag ctacaccgac gagctggccg agtgggttga atcccgcgcg
     4441 gccaagaagc gccggcgtga tgaggctgcg gttgcgttcc tggcggtgag ggcggatgtc
     4501 gaggcggcgt tagcgtccgg ctatgcgctc gtcaccattt gggagcacat gcgggaaacg
     4561 gggaaggtca agttctccta cgagacgttc cgctcgcacg ccaggcggca catcaaggcc
     4621 aagcccgccg atgtgcccgc accgcaggcc aaggctgcgg aacccgcgcc ggcacccaag
     4681 acgccggagc cacggcggcc gaagcagggg ggcaaggctg aaaagccggc ccccgctgcg
     4741 gccccgaccg gcttcacctt caacccaaca ccggacaaaa aggatcctct acgccggacg
     4801 catcgtggcc ggcatcaccg gcgccacaag tgcggttgct ggcgcctata tcgccgacat
     4861 caccgatggg gaagatcggg ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg
     4921 tatggtggca ggccccgtgg ccgggggact gttgggcgcc atctccttgc tgcctcgcgc
     4981 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     5041 gtctgtaagc ggatgccgta agtaaggtaa aggcctcgcg gaacccctat ttgtttattt
     5101 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5161 taatattgaa aaaggagctt atcaccgata aggaggtttt ttaatgacgg tgctgcactc
     5221 ggttgacttc ttccctagcg gcaatgccag cgttgccatt gagccgcgcc tgcctcaagc
     5281 cgacttcccg gagcaccacc acgacttcca cgagatcgtt atcgtggagc acggtaccgg
     5341 catccacgtt ttcaacggcc aaccgtacac gattacgggc ggtaccgtgt gcttcgttcg
     5401 tgatcacgac cgccacttat acgagcacac ggacaactta tgcttaacca acgttttata
     5461 ccgtagccct gaccgcttcc aattcctggc gggcttaaac caactgttac cgcaggaatt
     5521 agacggccaa taccctagcc attggcgtgt gaatcattcg gtgctgcaac aagttcgcca
     5581 attagtggcg caaatggagc aacaagaggg cgagaacgac ctgccgagca cggcgagccg
     5641 tgaaattctg ttcatgcagc tgttactgct gttacgcaag tcgagcctgc aagaaaattt
     5701 agagaattcg gcgagccgcc tgaatctgct gttagcgtgg ttagaagatc acttcgcgga
     5761 cgaagttaac tgggatgcgg ttgccgacca gttcagcctg agcttacgca ccctgcaccg
     5821 ccaactgaaa caacagaccg gcttaacccc gcaacgctat ttaaaccgtt tacgcttaat
     5881 gaaggcgcgc cacttactgc gccattcgga agcgtcggtg accgatattg cgtaccactg
     5941 tggcttttcg gatagcaatc atttcagcac cctgttccgt cgcgaattca attggagccc
     6001 tcgcgacatc cgccaaggcc gcgacggttt cttacagtga aggccttcta gagtcgacct
     6061 gcatcgacct gcagttgaca attaatcatc ggctcgtata atgtgtggaa aggcggttca
     6121 ccgccgtttt cacacaggaa acagaattct ttaagaagga gatatacata tgtcagtgat
     6181 agcaaagcag atgacataca aagtatatat gagcggtact gtaaatggtc actacttcga
     6241 agtagaaggt gatggcaaag ggaagccgta tgagggtgaa caaacagtga agcttacagt
     6301 tacgaagggt ggccctttgc cgttcgcgtg ggacattttg tcgccacagt gccagtacgg
     6361 gagcatacca ttcaccaagt atccagagga cataccagac tacgtgaaac agtcctttcc
     6421 cgaaggctat acctgggagc gcataatgaa ctttgaagac ggtgcggttt gtacggtatc
     6481 gaacgattca tcaatccaag gaaattgctt tatttatcat gttaaatttt cggggcttaa
     6541 ctttccgcca aatggccccg tgatgcagaa gaaaactcag ggctgggaac caaataccga
     6601 gcgccttttc gctcgggacg gaatgctgtt gggaaacaat tttatggcgt tgaagctgga
     6661 agggggtggc cactatctct gtgaattcaa gacaacatac aaagccaaga agcctgtcaa
     6721 gatgcccggt tatcactatg tggaccggaa gctcgatgtc actaatcaca acaaggatta
     6781 tacttcagtt gaacagtgcg aaatttcgat cgctcgcaaa cctgtcgtag cataagg
//
