LOCUS       40924_30550        8021 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pME6030, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8021)
  AUTHORS   Heeb S, Itoh Y, Nishijyo T, Schnider U, Keel C, Wade J, Walsh U, 
            O'Gara F, Haas D.
  TITLE     Small, stable shuttle vectors based on the minimal pVS1 replicon for
            use in gram-negative, plant-associated bacteria
  JOURNAL   Mol. Plant Microbe Interact. 13 (2), 232-237 (2000)
  PUBMED    10659714
REFERENCE   2  (bases 1 to 8021)
  AUTHORS   Heeb S.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-MAR-1999) Laboratoire de Biologie Microbienne, 
            University of Lausanne, Batiment de Biologie, Lausanne CH-1015, 
            Switzerland
REFERENCE   3  (bases 1 to 8021)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8021)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol. Plant 
            Microbe Interact."; date: "2000"; volume: "13"; issue: "2"; pages: 
            "232-237"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (10-MAR-1999) Laboratoire de Biologie Microbienne, University of 
            Lausanne, Batiment de Biologie, Lausanne CH-1015, Switzerland"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8021
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             195..881
                     /codon_start=1
                     /product="pVS1 resolvase"
                     /label=pVS1 resolvase
                     /note="ORF1; not required for stable maintenance"
                     /protein_id="AAD21668.1"
                     /translation="MNKSAAAGLLGYARVSTDDQDLTNQRAELHAAGCTKLFSEKITGT
                     RRDRPELARMLDHLRPGDVVTVTRLDRLARSTRDLLDIAERIQEAGAGLRSLAEPWADT
                     TTPAGRMVLTVFAGIAEFERSLIIDRTRSGREAAKARGVKFGPRPTLTPAQIAHARELI
                     DQEGRTVKEAAALLGVHRSTLYRALERSEEVTPTEARRRGAFREDALTEADALAAAENE
                     RQEEQA"
     CDS             878..1093
                     /codon_start=1
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="ORF2; not required for stable maintenance"
                     /protein_id="AAD21670.1"
                     /translation="MKPHQDGQDEPFFITEEIEAEMIAAGYVFEPPAHVSTVRLHEILA
                     GLSDAKLAAWPASLAAEETERRRLKR"
     CDS             1180..1806
                     /codon_start=1
                     /label=pVS1 StaA
                     /note="stability protein from the Pseudomonas plasmid pVS1
                     (Heeb et al., 2000)"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             1830..2045
                     /codon_start=1
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="ORF3; not required for stable maintenance"
                     /protein_id="AAD21671.1"
                     /translation="MSKSTNTLSAGRPSARSSKAATLASLADTPAMKRVNFQLPAEDHT
                     KLKMYAVRQGKTITELLSEYIAQLPE"
     misc_feature    2076..2088
                     /label=KorB box
                     /note="KorB box"
     CDS             2238..3308
                     /codon_start=1
                     /label=pVS1 RepA
                     /note="replication protein from the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
                     /translation="VSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      3377..3571
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     rep_origin      4281..4826
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     misc_feature    5068..5089
                     /label=p15A origin of transfer
                     /note="p15A origin of transfer"
     misc_feature    5137..5247
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             complement(6039..6686)
                     /codon_start=1
                     /label=TetR
                     /note="tetracycline resistance regulatory protein"
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     CDS             6792..7988
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="VKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
ORIGIN
        1 ggatccaacc cctccgctgc tatagtgcag tcggcttctg acgttcagtg cagccgtctt
       61 ctgaaaacga catgtcgcac aagtcctaag ttacgcgaca ggctgccgcc ctgccctttt
      121 cctggcgttt tcttgtcgcg tgttttagtc gcataaagta gaatacttgc gactagaacc
      181 ggagacatta cgccatgaac aagagcgccg ccgctggcct gctgggctat gcccgcgtca
      241 gcaccgacga ccaggacttg accaaccaac gggccgaact gcacgcggcc ggctgcacca
      301 agctgttttc cgagaagatc accggcacca ggcgcgaccg cccggagctg gccaggatgc
      361 ttgaccacct acgccctggc gacgttgtga cagtgaccag gctagaccgc ctggcccgca
      421 gcacccgcga cctactggac attgccgagc gcatccagga ggccggcgcg ggcctgcgta
      481 gcctggcaga gccgtgggcc gacaccacca cgccggccgg ccgcatggtg ttgaccgtgt
      541 tcgccggcat tgccgagttc gagcgttccc taatcatcga ccgcacccgg agcgggcgcg
      601 aggccgccaa ggcccgaggc gtgaagtttg gcccccgccc taccctcacc ccggcacaga
      661 tcgcgcacgc ccgcgagctg atcgaccagg aaggccgcac cgtgaaagag gcggctgcac
      721 tgcttggcgt gcatcgctcg accctgtacc gcgcacttga gcgcagcgag gaagtgacgc
      781 ccaccgaggc caggcggcgc ggtgccttcc gtgaggacgc attgaccgag gccgacgccc
      841 tggcggccgc cgagaatgaa cgccaagagg aacaagcatg aaaccgcacc aggacggcca
      901 ggacgaaccg tttttcatta ccgaagagat cgaggcggag atgatcgcgg ccgggtacgt
      961 gttcgagccg cccgcgcacg tctcaaccgt gcggctgcat gaaatcctgg ccggtttgtc
     1021 tgatgccaag ctggcggcct ggccggccag cttggccgct gaagaaaccg agcgccgccg
     1081 tctaaaaagg tgatgtgtat ttgagtaaaa cagcttgcgt catgcggtcg ctgcgtatat
     1141 gatgcgatga gtaaataaac aaatacgcaa ggggaacgca tgaaggttat cgctgtactt
     1201 aaccagaaag gcgggtcagg caagacgacc atcgcaaccc atctagcccg cgccctgcaa
     1261 ctcgccgggg ccgatgttct gttagtcgat tccgatcccc agggcagtgc ccgcgattgg
     1321 gcggccgtgc gggaagatca accgctaacc gttgtcggca tcgaccgccc gacgattgac
     1381 cgcgacgtga aggccatcgg ccggcgcgac ttcgtagtga tcgacggagc gccccaggcg
     1441 gcggacttgg ctgtgtccgc gatcaaggca gccgacttcg tgctgattcc ggtgcagcca
     1501 agcccttacg acatatgggc caccgccgac ctggtggagc tggttaagca gcgcattgag
     1561 gtcacggatg gaaggctaca agcggccttt gtcgtgtcgc gggcgatcaa aggcacgcgc
     1621 atcggcggtg aggttgccga ggcgctggcc gggtacgagc tgcccattct tgagtcccgt
     1681 atcacgcagc gcgtgagcta cccaggcact gccgccgccg gcacaaccgt tcttgaatca
     1741 gaacccgagg gcgacgctgc ccgcgaggtc caggcgctgg ccgctgaaat taaatcaaaa
     1801 ctcatttgag ttaatgaggt aaagagaaaa tgagcaaaag cacaaacacg ctaagtgccg
     1861 gccgtccgag cgcacgcagc agcaaggctg caacgttggc cagcctggca gacacgccag
     1921 ccatgaagcg ggtcaacttt cagttgccgg cggaggatca caccaagctg aagatgtacg
     1981 cggtacgcca aggcaagacc attaccgagc tgctatctga atacatcgcg cagctaccag
     2041 agtaaatgag caaatgaata aatgagtaga tgaattttag cggctaaagg aggcggcatg
     2101 gaaaatcaag aacaaccagg caccgacgcc gtggaatgcc ccatgtgtgg aggaacgggc
     2161 ggttggccag gcgtaagcgg ctgggttgtc tgccggccct gcaatggcac tggaaccccc
     2221 aagcccgagg aatcggcgtg agcggtcgca aaccatccgg cccggtacaa atcggcgcgg
     2281 cgctgggtga tgacctggtg gagaagttga aggccgcgca ggccgcccag cggcaacgca
     2341 tcgaggcaga agcacgcccc ggtgaatcgt ggcaagcggc cgctgatcga atccgcaaag
     2401 aatcccggca accgccggca gccggtgcgc cgtcgattag gaagccgccc aagggcgacg
     2461 agcaaccaga ttttttcgtt ccgatgctct atgacgtggg cacccgcgat agtcgcagca
     2521 tcatggacgt ggccgttttc cgtctgtcga agcgtgaccg acgagctggc gaggtgatcc
     2581 gctacgagct tccagacggg cacgtagagg tttccgcagg gccggccggc atggccagtg
     2641 tgtgggatta cgacctggta ctgatggcgg tttcccatct aaccgaatcc atgaaccgat
     2701 accgggaagg gaagggagac aagcccggcc gcgtgttccg tccacacgtt gcggacgtac
     2761 tcaagttctg ccggcgagcc gatggcggaa agcagaaaga cgacctggta gaaacctgca
     2821 ttcggttaaa caccacgcac gttgccatgc agcgtacgaa gaaggccaag aacggccgcc
     2881 tggtgacggt atccgagggt gaagccttga ttagccgcta caagatcgta aagagcgaaa
     2941 ccgggcggcc ggagtacatc gagatcgagc tagctgattg gatgtaccgc gagatcacag
     3001 aaggcaagaa cccggacgtg ctgacggttc accccgatta ctttttgatc gatcccggca
     3061 tcggccgttt tctctaccgc ctggcacgcc gcgccgcagg caaggcagaa gccagatggt
     3121 tgttcaagac gatctacgaa cgcagtggca gcgccggaga gttcaagaag ttctgtttca
     3181 ccgtgcgcaa gctgatcggg tcaaatgacc tgccggagta cgatttgaag gaggaggcgg
     3241 ggcaggctgg cccgatccta gtcatgcgct accgcaacct gatcgagggc gaagcatccg
     3301 ccggttccta atgtacggag cagatgctag ggcaaattgc cctagcaggg gaaaaaggtc
     3361 gaaaaggtct ctttcctgtg gatagcacgt acattgggaa cccaaagccg tacattggga
     3421 accggaaccc gtacattggg aacccaaagc cgtacattgg gaaccggtca cacatgtaag
     3481 tgactgatat aaaagagaaa aaaggcgatt tttccgccta aaactcttta aaacttatta
     3541 aaactcttaa aacccgcctg gcctgtgcat aactgtctgg ccagcgcaca gccgaagagc
     3601 tgcaaaaagc gcctaccctt cggtcgctgc gctccctacg ccccgccgct tcgcgtcggc
     3661 ctatcgcggc cgctggccgc tcaaaaatgg ctggcctacg gccaggcaat ctaccagggc
     3721 gcggacaagc cgcgccgtcg ccactcgacc gccggcgctg aggtctgcct cgtgaagaag
     3781 gtgttgctga ctcataccag ggcaatggca acaacgttgc gcaaactatt aactggcgaa
     3841 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     3901 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     3961 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     4021 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     4081 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     4141 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     4201 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     4261 cccttaataa gatgatcttc ttgagatcgt tttggtctgc gcgtaatctc ttgctctgaa
     4321 aacgaaaaaa ccgccttgca gggcggtttt tcgaaggttc tctgagctac caactctttg
     4381 aaccgaggta actggcttgg aggagcgcag tcaccaaaac ttgtcctttc agtttagcct
     4441 taaccggcgc atgacttcaa gactaactcc tctaaatcaa ttaccagtgg ctgctgccag
     4501 tggtgctttt gcatgtcttt ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4561 gcggtcggac tgaacggggg gttcgtgcat acagtccagc ttggagcgaa ctgcctaccc
     4621 ggaactgagt gtcaggcgtg gaatgagaca aacgcggcca taacagcgga atgacaccgg
     4681 taaaccgaaa ggcaggaaca ggagagcgca cgagggagcc gccaggggga aacgcctggt
     4741 atctttatag tcctgtcggg tttcgccacc actgatttga gcgtcagatt tcgtgatgct
     4801 tgtcaggggg gcggagccta tggaaaaacg gctttgccgc ggccctctca cttccctgtt
     4861 aagtatcttc ctggcatctt ccaggaaatc tccgccccgt tcgtaagcca tttccgctcg
     4921 ccgcagtcga acgaccgagc gtagcgagtc agtgagcgag gaagcggaat atatcctgta
     4981 tcacatattc tgctgacgca ccggtgcagc cttttttctc ctgccacatg aagcacttca
     5041 ctgacaccct catcagtgcc aacatagtaa gccagtatac actccgctag cgctgaggtc
     5101 tgcctcgtga agaaggtgtt gctgactcat accaggctag tccgaggcct cgagatctat
     5161 cgatgcatgc catggtaccc gggagctcga attcgaagct tctgcagacg cgtcgacgtc
     5221 atatggatcc gatatcgccg tggcggcttt tgccattctc accggattca gtcgtcactc
     5281 atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg
     5341 atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc
     5401 tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc
     5461 ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg
     5521 ttaattggtt gtaacactgg cagagcatta cgctgacttg acgggacggc ggctttgttg
     5581 aataaatcga acttttgctg agttgaagga tcagatcacg catcttcccg acaacgcaga
     5641 ccgttccgtg gcaaagcaaa agttcaaaat caccaactgg tccacctaca acaaagctct
     5701 catcaaccgt ggctccctca ctttctggct ggatgatggg gcgattcagg cctggtatga
     5761 gtcagcaaca ccttcttcac gaggcagacc tcagcgccag gcgcacgagg ccggcgcgcg
     5821 gacccgcgcc acggcccacg agcggacgcc gcagcaggag cgccagaagg ccgccagaga
     5881 ggccgagcgc ggccgtgagg cttggacgct agggcagggc atgaaaaagc ccgtagcggg
     5941 ctgctacggg cgtctgacgc ggtggaaagg gggaggggat gttgtctaca tggctctgct
     6001 gtagtgagtg ggttgcgctc cggcagcggt cctgatcaat cgtcaccctt tctcggtcct
     6061 tcaacgttcc tgacaacgag cctccttttc gccaatccat cgacaatcac cgcgagtccc
     6121 tgctcgaacg ctgcgtccgg accggcttcg tcgaaggcgt ctatcgcggc ccgcaacagc
     6181 ggcgagagcg gagcctgttc aacggtgccg ccgcgctcgc cggcatcgct gtcgccggcc
     6241 tgctcctcaa gcacggcccc aacagtgaag tagctgattg tcatcagcgc attgacggcg
     6301 tccccggccg aaaaacccgc ctcgcagagg aagcgaagct gcgcgtcggc cgtttccatc
     6361 tgcggtgcgc ccggtcgcgt gccggcatgg atgcgcgcgc catcgcggta ggcgagcagc
     6421 gcctgcctga agctgcgggc attcccgatc agaaatgagc gccagtcgtc gtcggctctc
     6481 ggcaccgaat gcgtatgatt ctccgccagc atggcttcgg ccagtgcgtc gagcagcgcc
     6541 cgcttgttcc tgaagtgcca gtaaagcgcc ggctgctgaa cccccaaccg ttccgccagt
     6601 ttgcgtgtcg tcagaccgtc tacgccgacc tcgttcaaca ggtccagggc ggcacggatc
     6661 actgtattcg gctgcaactt tgtcatgctt gacactttat cactgataaa cataatatgt
     6721 ccaccaactt atcagtgata aagaatccgc gcgttcaatc ggaccagcgg aggctggtcc
     6781 ggaggccaga cgtgaaaccc aacatacccc tgatcgtaat tctgagcact gtcgcgctcg
     6841 acgctgtcgg catcggcctg attatgccgg tgctgccggg cctcctgcgc gatctggttc
     6901 actcgaacga cgtcaccgcc cactatggca ttctgctggc gctgtatgcg ttggtgcaat
     6961 ttgcctgcgc acctgtgctg ggcgcgctgt cggatcgttt cgggcggcgg ccaatcttgc
     7021 tcgtctcgct ggccggcgcc actgtcgact acgccatcat ggcgacagcg cctttccttt
     7081 gggttctcta tatcgggcgg atcgtggccg gcatcaccgg ggcgactggg gcggtagccg
     7141 gcgcttatat tgccgatatc actgatggcg atgagcgcgc gcggcacttc ggcttcatga
     7201 gcgcctgttt cgggttcggg atggtcgcgg gacctgtgct cggtgggctg atgggcggtt
     7261 tctcccccca cgctccgttc ttcgccgcgg cagccttgaa cggcctcaat ttcctgacgg
     7321 gctgtttcct tttgccggag tcgcacaaag gcgaacgccg gccgttacgc cgggaggctc
     7381 tcaacccgct cgcttcgttc cggtgggccc ggggcatgac cgtcgtcgcc gccctgatgg
     7441 cggtcttctt catcatgcaa cttgtcggac aggtgccggc cgcgctttgg gtcattttcg
     7501 gcgaggatcg ctttcactgg gacgcgacca cgatcggcat ttcgcttgcc gcatttggca
     7561 ttctgcattc actcgcccag gcaatgatca ccggccctgt agccgcccgg ctcggcgaaa
     7621 ggcgggcact catgctcgga atgattgccg acggcacagg ctacatcctg cttgccttcg
     7681 cgacacgggg atggatggcg ttcccgatca tggtcctgct tgcttcgggt ggcatcggaa
     7741 tgccggcgct gcaagcaatg ttgtccaggc aggtggatga ggaacgtcag gggcagctgc
     7801 aaggctcact ggcggcgctc accagcctga cctcgatcgt cggacccctc ctcttcacgg
     7861 cgatctatgc ggcttctata acaacgtgga acgggtgggc atggattgca ggcgctgccc
     7921 tctacttgct ctgcctgccg gcgctgcgtc gcgggctttg gagcggcgca gggcaacgag
     7981 ccgatcgctg atcgtggaaa cgataggccg ctctagaact a
//
