LOCUS       40924_18721        7626 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pEX1.8 plasmid pEX1.8, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7626)
  AUTHORS   Pearson JP, Pesci EC, Iglewski BH.
  TITLE     Roles of Pseudomonas aeruginosa las and rhl quorum-sensing systems 
            in control of elastase and rhamnolipid biosynthesis genes
  JOURNAL   J. Bacteriol. 179 (18), 5756-5767 (1997)
  PUBMED    9294432
REFERENCE   2  (bases 1 to 7626)
  AUTHORS   Schwingel JM, Iglewski BH.
  TITLE     pEX1.8, Pseudomonas aeruginosa/E. coli shuttle vector
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 7626)
  AUTHORS   Schwingel JM, Iglewski BH.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-JAN-2012) Department of Microbiology and Immunology, 
            University of Rochester, 601 Elmwood Ave., Box 672, Rochester, NY 
            14642, USA
REFERENCE   4  (bases 1 to 7626)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 7626)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. 
            Bacteriol."; date: "1997"; volume: "179"; issue: "18"; pages: 
            "5756-5767"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (02-JAN-2012) Department of Microbiology and Immunology, University 
            of Rochester, 601 Elmwood Ave., Box 672, Rochester, NY 14642, USA"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7626
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1813..2164)
                     /direction=LEFT
                     /label=pRO1600 oriV
                     /note="broad-host-range origin of replication from
                     Pseudomonas aeruginosa plasmid pRO1600; requires the 
                     pRO1600 Rep protein for replication (West et al., 1994)"
     CDS             2178..3008
                     /codon_start=1
                     /label=pRO1600 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pRO1600 from Pseudomonas aeruginosa"
                     /translation="VASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     promoter        4120..4148
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    4156..4172
                     /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)."
     terminator      4427..4513
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      4605..4632
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        4651..4742
                     /label=AmpR promoter
     CDS             4743..5600
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5774..6362
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(6548..6688)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     promoter        6864..6941
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             join(6942..7626,1..362)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQL"
ORIGIN
        1 ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc aacgatcaga
       61 tggcgctggg cgcaatgcgc gccattaccg agtccgggct gcgcgttggt gcggatatct
      121 cggtagtggg atacgacgat accgaagaca gctcatgtta tatcccgccg ttaaccacca
      181 tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg caactctctc
      241 agggccaggc ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa agaaaaacca
      301 ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
      361 tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg
      421 cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca
      481 tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt
      541 tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg
      601 atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc
      661 gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg
      721 ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg
      781 ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg
      841 ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt
      901 cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca
      961 cgatcatgcg cacccgtggc caggacccaa cgctgcccga gatgcgccgc gtgcggctgc
     1021 tggagatggc ggacgcgatg gatatgttct gccaagggtt ggtttgcgca ttcacagttc
     1081 tccgcaagaa ttgattggct ccaattcttg gagtggtgaa tccgttagcg aggtgccgcc
     1141 ggcttccatt caggtcgagg gcaatggcaa caacgttgcg caaactatta actggcgaac
     1201 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     1261 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     1321 gtgagcgtgg atctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     1381 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     1441 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     1501 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     1561 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     1621 ccaattacac gccactggct gtgcttgctg gggtgacggt ggcaacggtg gcggccttgc
     1681 tgggctatcg cgttggaaag aaacgaggga aaggggactg ataaaccggt cttagcccct
     1741 ccccttggtg tccaaccgct ctgtaggcct ctcaggcgcc gctggtgccg ctggttggac
     1801 gccaagggtg aatccgcctc gataccctga ttactcgctt cctgcgccct ctcaggcggc
     1861 gataggggac tggtaaaacg gggattgccc agacgcctcc cccgcccctt caggggcaca
     1921 aatgcggccc caacggggcc acgtagtggt gcgttttttg cgtttccacc cttttcttcc
     1981 ttttcccttt taaacctttt aggacgtcta caggccacgt aatccgtggc ctgtagagtt
     2041 taaaaaggga cggatttgtt gccattaagg gacggatttg ttgttaagaa gggacggatt
     2101 tgttgttgta aagggacgga tttgttgtat tgtgggacgc agatacagtg tccccttata
     2161 cacaaggaat gtcgaacgtg gcctcacccc caatggttta caaaagcaat gccctggtcg
     2221 aggccgcgta tcgcctcagt gttcaggaac agcggatcgt tctggcctgt attagccagg
     2281 tgaagaggag cgagcctgtc accgatgaag tgatgtattc agtgacggcg gaggacatag
     2341 cgacgatggc gggtgtccct atcgaatctt cctacaacca gctcaaagaa gcggccctgc
     2401 gcctgaaacg gcgggaagtc cggttaaccc aagagcccaa tggcaagggg aaaagaccga
     2461 gtgtgatgat taccggctgg gtgcaaacaa tcatctaccg ggagggtgag ggccgtgtag
     2521 aactcaggtt caccaaagac atgctgccgt acctgacgga actcaccaaa cagttcacca
     2581 aatacgcctt ggctgacgtg gccaagatgg acagcaccca cgcgatcagg ctttacgagc
     2641 tgctcatgca atgggacagc atcggccagc gcgaaataga aattgaccag ctgcgaaagt
     2701 ggtttcaact ggaaggccgg tatccctcga tcaaggactt caagttgcga gtgcttgatc
     2761 cagccgtgac gcagatcaac gagcacagcc cgctacaggt ggagtgggcg cagcgaaaga
     2821 ccgggcgcaa ggtcacacat ctgttgttca gttttggacc gaagaagccc gccaaggcgg
     2881 tgggtaaggc cccagcgaag cgcaaggccg ggaagatttc agatgctgag atcgcgaaac
     2941 aggctcgccc tggtgagaca tgggaagcgg cccgcgctcg actaacccag atgccgctgg
     3001 atctggccta gaggccgtgg ccaccacggc ccggcctgcc tttcgtggcc cggctccatg
     3061 caccgcgacg caacgcgggg aggcagacaa ggtatagggc ggcgcctaca atccatgcca
     3121 acccgttcca tgtgctcgcc gaggcggcat aaatcgccgt gacgatcagc ggtccaatga
     3181 tcgaagttag gctggtaaga gccgcgagcg atccttgaag ctgtccctga tggtcgtcat
     3241 ctacctgcct ggacagcatg gcctgcaacg cgggcatccc gatgccgccg gaagcgagaa
     3301 gaatcataat ggggaaggcc atccagcctc gcgtcgcgaa cgccagcaag acgtagccca
     3361 gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg
     3421 gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc
     3481 cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg
     3541 gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca
     3601 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgac
     3661 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga
     3721 gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca
     3781 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc
     3841 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
     3901 tgatgccggc cacgatgcgt ccggcgtaga ggatccgggc ttatcgactg cacggtgcac
     3961 caatgcttct ggcgtcaggc agccatcgga agctgtggta tggctgtgca ggtcgtaaat
     4021 cactgcataa ttcgtgtcgc tcaaggcgca ctcccgttct ggataatgtt ttttgcgccg
     4081 acatcataac ggttctggca aatattctga aatgagctgt tgacaattaa tcatcggctc
     4141 gtataatgtg tggaattgtg agcggataac aatttcacac aggaaacaga attcccgggg
     4201 atccgtcgac ctgcagccaa gcttggctgt tttggcggat gagagaagat tttcagcctg
     4261 atacagatta aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt
     4321 agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat
     4381 ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa
     4441 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
     4501 gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg
     4561 gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac
     4621 ggatggcctt tttgcgtttc tacaaactct tttgtttatt tttctaaata cattcaaata
     4681 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4741 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4801 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4861 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4921 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4981 cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5041 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5101 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5161 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5221 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5281 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5341 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5401 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5461 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5521 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5581 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5641 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5701 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5761 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5821 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5881 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     5941 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6001 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6061 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6121 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6181 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6241 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6301 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6361 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6421 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     6481 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     6541 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat
     6601 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct
     6661 atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc
     6721 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     6781 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc gcgcgcgaag
     6841 gcgaagcggc atgcatttac gttgacacca tcgaatggtg caaaaccttt cgcggtatgg
     6901 catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat
     6961 acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg
     7021 ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt
     7081 acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg
     7141 ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg
     7201 ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct
     7261 gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc
     7321 cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat
     7381 ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat gaagacggta
     7441 cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg
     7501 gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc
     7561 gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc
     7621 aacaaa
//
