LOCUS       Exported                8243 bp DNA     circular SYN 20-JUL-2025
DEFINITION  Shuttle vector pME6031, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8243)
  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 8243)
  AUTHORS   Heeb S.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-DEC-1998) Laboratoire de Biologie Microbienne, 
            University of Lausanne, Batiment de Biologie, Lausanne CH-1015, 
            Switzerland
REFERENCE   3  (bases 1 to 8243)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8243)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 8243)
  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
            (30-DEC-1998) Laboratoire de Biologie Microbienne, University of
            Lausanne, Batiment de Biologie, Lausanne CH-1015, Switzerland"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8243
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             194..880
                     /codon_start=1
                     /product="pVS1 resolvase"
                     /label=pVS1 resolvase
                     /note="ORF1; not required for stable maintenance"
                     /protein_id="AAD19683.1"
                     /translation="MNKSAAAGLLGYARVSTDDQDLTNQRAELHAAGCTKLFSEKITGT
                     RRDRPELARMLDHLRPGDVVTVTRLDRLARSTRDLLDIAERIQEAGAGLRSLAEPWADT
                     TTPAGRMVLTVFAGIAEFERSLIIDRTRSGREAAKARGVKFGPRPTLTPAQIAHARELI
                     DQEGRTVKEAAALLGVHRSTLYRALERSEEVTPTEARRRGAFREDALTEADALAAAENE
                     RQEEQA"
     CDS             877..1092
                     /codon_start=1
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="ORF2; not required for stable maintenance"
                     /protein_id="AAD19684.1"
                     /translation="MKPHQDGQDEPFFITEEIEAEMIAAGYVFEPPAHVSTVRLHEILA
                     GLSDAKLAAWPASLAAEETERRRLKR"
     CDS             1179..1805
                     /label=pVS1 StaA
                     /note="stability protein from the Pseudomonas plasmid pVS1
                     (Heeb et al., 2000)"
     CDS             1829..2044
                     /codon_start=1
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="ORF3; not required for stable maintenance"
                     /protein_id="AAD19686.1"
                     /translation="MSKSTNTLSAGRPSARSSKAATLASLADTPAMKRVNFQLPAEDHT
                     KLKMYAVRQGKTITELLSEYIAQLPE"
     misc_feature    2074..2087
                     /label=KorB box
                     /note="KorB box"
     CDS             2237..3307
                     /label=pVS1 RepA
                     /note="replication protein from the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     rep_origin      3376..3570
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     rep_origin      4279..4824
                     /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    5066..5087
                     /label=p15A origin of transfer
                     /note="p15A origin of transfer"
     misc_difference 5134..5411
                     /label=part of terminator absent in pME6030
                     /note="part of terminator absent in pME6030"
     regulatory      5145..5411
                     /label=T4 transcription terminator
                     /note="T4 transcription terminator"
                     /regulatory_class="terminator"
     misc_feature    5413..5523
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             complement(6249..6896)
                     /label=TetR
                     /note="tetracycline resistance regulatory protein"
     CDS             7002..8198
                     /label=TcR
                     /note="tetracycline efflux protein"
ORIGIN
        1 gatccaaccc ctccgctgct atagtgcagt cggcttctga cgttcagtgc agccgtcttc
       61 tgaaaacgac atgtcgcaca agtcctaagt tacgcgacag gctgccgccc tgcccttttc
      121 ctggcgtttt cttgtcgcgt gttttagtcg cataaagtag aatacttgcg actagaaccg
      181 gagacattac gccatgaaca agagcgccgc cgctggcctg ctgggctatg cccgcgtcag
      241 caccgacgac caggacttga ccaaccaacg ggccgaactg cacgcggccg gctgcaccaa
      301 gctgttttcc gagaagatca ccggcaccag gcgcgaccgc ccggagctgg ccaggatgct
      361 tgaccaccta cgccctggcg acgttgtgac agtgaccagg ctagaccgcc tggcccgcag
      421 cacccgcgac ctactggaca ttgccgagcg catccaggag gccggcgcgg gcctgcgtag
      481 cctggcagag ccgtgggccg acaccaccac gccggccggc cgcatggtgt tgaccgtgtt
      541 cgccggcatt gccgagttcg agcgttccct aatcatcgac cgcacccgga gcgggcgcga
      601 ggccgccaag gcccgaggcg tgaagtttgg cccccgccct accctcaccc cggcacagat
      661 cgcgcacgcc cgcgagctga tcgaccagga aggccgcacc gtgaaagagg cggctgcact
      721 gcttggcgtg catcgctcga ccctgtaccg cgcacttgag cgcagcgagg aagtgacgcc
      781 caccgaggcc aggcggcgcg gtgccttccg tgaggacgca ttgaccgagg ccgacgccct
      841 ggcggccgcc gagaatgaac gccaagagga acaagcatga aaccgcacca ggacggccag
      901 gacgaaccgt ttttcattac cgaagagatc gaggcggaga tgatcgcggc cgggtacgtg
      961 ttcgagccgc ccgcgcacgt ctcaaccgtg cggctgcatg aaatcctggc cggtttgtct
     1021 gatgccaagc tggcggcctg gccggccagc ttggccgctg aagaaaccga gcgccgccgt
     1081 ctaaaaaggt gatgtgtatt tgagtaaaac agcttgcgtc atgcggtcgc tgcgtatatg
     1141 atgcgatgag taaataaaca aatacgcaag gggaacgcat gaaggttatc gctgtactta
     1201 accagaaagg cgggtcaggc aagacgacca tcgcaaccca tctagcccgc gccctgcaac
     1261 tcgccggggc cgatgttctg ttagtcgatt ccgatcccca gggcagtgcc cgcgattggg
     1321 cggccgtgcg ggaagatcaa ccgctaaccg ttgtcggcat cgaccgcccg acgattgacc
     1381 gcgacgtgaa ggccatcggc cggcgcgact tcgtagtgat cgacggagcg ccccaggcgg
     1441 cggacttggc tgtgtccgcg atcaaggcag ccgacttcgt gctgattccg gtgcagccaa
     1501 gcccttacga catatgggcc accgccgacc tggtggagct ggttaagcag cgcattgagg
     1561 tcacggatgg aaggctacaa gcggcctttg tcgtgtcgcg ggcgatcaaa ggcacgcgca
     1621 tcggcggtga ggttgccgag gcgctggccg ggtacgagct gcccattctt gagtcccgta
     1681 tcacgcagcg cgtgagctac ccaggcactg ccgccgccgg cacaaccgtt cttgaatcag
     1741 aacccgaggg cgacgctgcc cgcgaggtcc aggcgctggc cgctgaaatt aaatcaaaac
     1801 tcatttgagt taatgaggta aagagaaaat gagcaaaagc acaaacacgc taagtgccgg
     1861 ccgtccgagc gcacgcagca gcaaggctgc aacgttggcc agcctggcag acacgccagc
     1921 catgaagcgg gtcaactttc agttgccggc ggaggatcac accaagctga agatgtacgc
     1981 ggtacgccaa ggcaagacca ttaccgagct gctatctgaa tacatcgcgc agctaccaga
     2041 gtaaatgagc aaatgaataa atgagtagat gaattttagc ggctaaagga ggcggcatgg
     2101 aaaatcaaga acaaccaggc accgacgccg tggaatgccc catgtgtgga ggaacgggcg
     2161 gttggccagg cgtaagcggc tgggttgtct gccggccctg caatggcact ggaaccccca
     2221 agcccgagga atcggcgtga gcggtcgcaa accatccggc ccggtacaaa tcggcgcggc
     2281 gctgggtgat gacctggtgg agaagttgaa ggccgcgcag gccgcccagc ggcaacgcat
     2341 cgaggcagaa gcacgccccg gtgaatcgtg gcaagcggcc gctgatcgaa tccgcaaaga
     2401 atcccggcaa ccgccggcag ccggtgcgcc gtcgattagg aagccgccca agggcgacga
     2461 gcaaccagat tttttcgttc cgatgctcta tgacgtgggc acccgcgata gtcgcagcat
     2521 catggacgtg gccgttttcc gtctgtcgaa gcgtgaccga cgagctggcg aggtgatccg
     2581 ctacgagctt ccagacgggc acgtagaggt ttccgcaggg ccggccggca tggccagtgt
     2641 gtgggattac gacctggtac tgatggcggt ttcccatcta accgaatcca tgaaccgata
     2701 ccgggaaggg aagggagaca agcccggccg cgtgttccgt ccacacgttg cggacgtact
     2761 caagttctgc cggcgagccg atggcggaaa gcagaaagac gacctggtag aaacctgcat
     2821 tcggttaaac accacgcacg ttgccatgca gcgtacgaag aaggccaaga acggccgcct
     2881 ggtgacggta tccgagggtg aagccttgat tagccgctac aagatcgtaa agagcgaaac
     2941 cgggcggccg gagtacatcg agatcgagct agctgattgg atgtaccgcg agatcacaga
     3001 aggcaagaac ccggacgtgc tgacggttca ccccgattac tttttgatcg atcccggcat
     3061 cggccgtttt ctctaccgcc tggcacgccg cgccgcaggc aaggcagaag ccagatggtt
     3121 gttcaagacg atctacgaac gcagtggcag cgccggagag ttcaagaagt tctgtttcac
     3181 cgtgcgcaag ctgatcgggt caaatgacct gccggagtac gatttgaagg aggaggcggg
     3241 gcaggctggc ccgatcctag tcatgcgcta ccgcaacctg atcgagggcg aagcatccgc
     3301 cggttcctaa tgtacggagc agatgctagg gcaaattgcc ctagcagggg aaaaaggtcg
     3361 aaaaggtctc tttcctgtgg atagcacgta cattgggaac ccaaagccgt acattgggaa
     3421 ccggaacccg tacattggga acccaaagcc gtacattggg aaccggtcac acatgtaagt
     3481 gactgatata aaagagaaaa aaggcgattt ttccgcctaa aactctttaa aacttattaa
     3541 aactcttaaa acccgcctgg cctgtgcata actgtctggc cagcgcacag ccgaagagct
     3601 gcaaaaagcg cctacccttc ggtcgctgcg ctccctacgc cccgccgctt cgcgtcggcc
     3661 tatcgcggcc gctggccgct caaaaatggc tggcctacgg ccaggcaatc taccagggcg
     3721 cggacaagcc gcgccgtcgc cactcgaccg ccggcgctga ggtctgcctc gtgaagaagg
     3781 tgttgctgac tcataccagg caatggcaac aacgttgcgc aaactattaa ctggcgaact
     3841 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     3901 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     3961 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4021 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4081 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4141 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4201 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4261 cttaataaga tgatcttctt gagatcgttt tggtctgcgc gtaatctctt gctctgaaaa
     4321 cgaaaaaacc gccttgcagg gcggtttttc gaaggttctc tgagctacca actctttgaa
     4381 ccgaggtaac tggcttggag gagcgcagtc accaaaactt gtcctttcag tttagcctta
     4441 accggcgcat gacttcaaga ctaactcctc taaatcaatt accagtggct gctgccagtg
     4501 gtgcttttgc atgtctttcc gggttggact caagacgata gttaccggat aaggcgcagc
     4561 ggtcggactg aacggggggt tcgtgcatac agtccagctt ggagcgaact gcctacccgg
     4621 aactgagtgt caggcgtgga atgagacaaa cgcggccata acagcggaat gacaccggta
     4681 aaccgaaagg caggaacagg agagcgcacg agggagccgc cagggggaaa cgcctggtat
     4741 ctttatagtc ctgtcgggtt tcgccaccac tgatttgagc gtcagatttc gtgatgcttg
     4801 tcaggggggc ggagcctatg gaaaaacggc tttgccgcgg ccctctcact tccctgttaa
     4861 gtatcttcct ggcatcttcc aggaaatctc cgccccgttc gtaagccatt tccgctcgcc
     4921 gcagtcgaac gaccgagcgt agcgagtcag tgagcgagga agcggaatat atcctgtatc
     4981 acatattctg ctgacgcacc ggtgcagcct tttttctcct gccacatgaa gcacttcact
     5041 gacaccctca tcagtgccaa catagtaagc cagtatacac tccgctagcg ctgaggtctg
     5101 cctcgtgaag aaggtgttgc tgactcatac caggtagtcc gaggagcttt atgcttgtaa
     5161 accgttttgt gaaaaaattt ttaaaataaa aaaggggacc tctagggtcc ccaattaatt
     5221 agtaatataa tctattaaag gtcattcaaa aggtcatcca ccggatcaat tcccctgctc
     5281 gcgcaggctg ggtgccaagc tctcgggtaa catcaaggcc cgatccttgg agcccttgcc
     5341 ctcccgcacg atgatcgtgc cgtgatcgaa atccagatcc ttgacccgca gttgcaaacc
     5401 ctcactgatc cgctagtccg aggcctcgag atctatcgat gcatgccatg gtacccggga
     5461 gctcgaattc gaagcttctg cagacgcgtc gacgtcatat ggatccgata tcgccgtggc
     5521 ggcttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc
     5581 ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag
     5641 accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac
     5701 agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc
     5761 atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga
     5821 gcattacgct gacttgacgg gacggcggct ttgttgaata aatcgaactt ttgctgagtt
     5881 gaaggatcag atcacgcatc ttcccgacaa cgcagaccgt tccgtggcaa agcaaaagtt
     5941 caaaatcacc aactggtcca cctacaacaa agctctcatc aaccgtggct ccctcacttt
     6001 ctggctggat gatggggcga ttcaggcctg gtatgagtca gcaacacctt cttcacgagg
     6061 cagacctcag cgccagaagg ccgccagaga ggccgagcgc ggccgtgagg cttggacgct
     6121 agggcagggc atgaaaaagc ccgtagcggg ctgctacggg cgtctgacgc ggtggaaagg
     6181 gggaggggat gttgtctaca tggctctgct gtagtgagtg ggttgcgctc cggcagcggt
     6241 cctgatcaat cgtcaccctt tctcggtcct tcaacgttcc tgacaacgag cctccttttc
     6301 gccaatccat cgacaatcac cgcgagtccc tgctcgaacg ctgcgtccgg accggcttcg
     6361 tcgaaggcgt ctatcgcggc ccgcaacagc ggcgagagcg gagcctgttc aacggtgccg
     6421 ccgcgctcgc cggcatcgct gtcgccggcc tgctcctcaa gcacggcccc aacagtgaag
     6481 tagctgattg tcatcagcgc attgacggcg tccccggccg aaaaacccgc ctcgcagagg
     6541 aagcgaagct gcgcgtcggc cgtttccatc tgcggtgcgc ccggtcgcgt gccggcatgg
     6601 atgcgcgcgc catcgcggta ggcgagcagc gcctgcctga agctgcgggc attcccgatc
     6661 agaaatgagc gccagtcgtc gtcggctctc ggcaccgaat gcgtatgatt ctccgccagc
     6721 atggcttcgg ccagtgcgtc gagcagcgcc cgcttgttcc tgaagtgcca gtaaagcgcc
     6781 ggctgctgaa cccccaaccg ttccgccagt ttgcgtgtcg tcagaccgtc tacgccgacc
     6841 tcgttcaaca ggtccagggc ggcacggatc actgtattcg gctgcaactt tgtcatgctt
     6901 gacactttat cactgataaa cataatatgt ccaccaactt atcagtgata aagaatccgc
     6961 gcgttcaatc ggaccagcgg aggctggtcc ggaggccaga cgtgaaaccc aacatacccc
     7021 tgatcgtaat tctgagcact gtcgcgctcg acgctgtcgg catcggcctg attatgccgg
     7081 tgctgccggg cctcctgcgc gatctggttc actcgaacga cgtcaccgcc cactatggca
     7141 ttctgctggc gctgtatgcg ttggtgcaat ttgcctgcgc acctgtgctg ggcgcgctgt
     7201 cggatcgttt cgggcggcgg ccaatcttgc tcgtctcgct ggccggcgcc actgtcgact
     7261 acgccatcat ggcgacagcg cctttccttt gggttctcta tatcgggcgg atcgtggccg
     7321 gcatcaccgg ggcgactggg gcggtagccg gcgcttatat tgccgatatc actgatggcg
     7381 atgagcgcgc gcggcacttc ggcttcatga gcgcctgttt cgggttcggg atggtcgcgg
     7441 gacctgtgct cggtgggctg atgggcggtt tctcccccca cgctccgttc ttcgccgcgg
     7501 cagccttgaa cggcctcaat ttcctgacgg gctgtttcct tttgccggag tcgcacaaag
     7561 gcgaacgccg gccgttacgc cgggaggctc tcaacccgct cgcttcgttc cggtgggccc
     7621 ggggcatgac cgtcgtcgcc gccctgatgg cggtcttctt catcatgcaa cttgtcggac
     7681 aggtgccggc cgcgctttgg gtcattttcg gcgaggatcg ctttcactgg gacgcgacca
     7741 cgatcggcat ttcgcttgcc gcatttggca ttctgcattc actcgcccag gcaatgatca
     7801 ccggccctgt agccgcccgg ctcggcgaaa ggcgggcact catgctcgga atgattgccg
     7861 acggcacagg ctacatcctg cttgccttcg cgacacgggg atggatggcg ttcccgatca
     7921 tggtcctgct tgcttcgggt ggcatcggaa tgccggcgct gcaagcaatg ttgtccaggc
     7981 aggtggatga ggaacgtcag gggcagctgc aaggctcact ggcggcgctc accagcctga
     8041 cctcgatcgt cggacccctc ctcttcacgg cgatctatgc ggcttctata acaacgtgga
     8101 acgggtgggc atggattgca ggcgctgccc tctacttgct ctgcctgccg gcgctgcgtc
     8161 gcgggctttg gagcggcgca gggcaacgag ccgatcgctg atcgtggaaa cgataggggc
     8221 cgctctagaa ctagctagaa cta
//
