LOCUS       40924_30565        5303 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pME6040, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5303)
  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 5303)
  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 5303)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5303)
  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..5303
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             195..821
                     /codon_start=1
                     /label=pVS1 StaA
                     /note="stability protein from the Pseudomonas plasmid pVS1
                     (Heeb et al., 2000)"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             845..1060
                     /codon_start=1
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="ORF3; not required for stable maintenance"
                     /protein_id="AAD21674.1"
                     /translation="MSKSTNTLSAGRPSARSSKAATLASLADTPAMKRVNFQLPAEDHT
                     KLKMYAVRQGKTITELLSEYIAQLPE"
     misc_feature    1091..1103
                     /label=KorB box
                     /note="KorB box"
     CDS             1253..2323
                     /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      2392..2586
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     rep_origin      2981..3526
                     /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    3768..3789
                     /label=p15A origin of transfer
                     /note="p15A origin of transfer"
     misc_feature    3837..3947
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             4229..5041
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
ORIGIN
        1 accgtgcggc tgcatgaaat cctggccggt ttgtctgatg ccaagctggc ggcctggccg
       61 gccagcttgg ccgctgaaga aaccgagcgc cgccgtctaa aaaggtgatg tgtatttgag
      121 taaaacagct tgcgtcatgc ggtcgctgcg tatatgatgc gatgagtaaa taaacaaata
      181 cgcaagggga acgcatgaag gttatcgctg tacttaacca gaaaggcggg tcaggcaaga
      241 cgaccatcgc aacccatcta gcccgcgccc tgcaactcgc cggggccgat gttctgttag
      301 tcgattccga tccccagggc agtgcccgcg attgggcggc cgtgcgggaa gatcaaccgc
      361 taaccgttgt cggcatcgac cgcccgacga ttgaccgcga cgtgaaggcc atcggccggc
      421 gcgacttcgt agtgatcgac ggagcgcccc aggcggcgga cttggctgtg tccgcgatca
      481 aggcagccga cttcgtgctg attccggtgc agccaagccc ttacgacata tgggccaccg
      541 ccgacctggt ggagctggtt aagcagcgca ttgaggtcac ggatggaagg ctacaagcgg
      601 cctttgtcgt gtcgcgggcg atcaaaggca cgcgcatcgg cggtgaggtt gccgaggcgc
      661 tggccgggta cgagctgccc attcttgagt cccgtatcac gcagcgcgtg agctacccag
      721 gcactgccgc cgccggcaca accgttcttg aatcagaacc cgagggcgac gctgcccgcg
      781 aggtccaggc gctggccgct gaaattaaat caaaactcat ttgagttaat gaggtaaaga
      841 gaaaatgagc aaaagcacaa acacgctaag tgccggccgt ccgagcgcac gcagcagcaa
      901 ggctgcaacg ttggccagcc tggcagacac gccagccatg aagcgggtca actttcagtt
      961 gccggcggag gatcacacca agctgaagat gtacgcggta cgccaaggca agaccattac
     1021 cgagctgcta tctgaataca tcgcgcagct accagagtaa atgagcaaat gaataaatga
     1081 gtagatgaat tttagcggct aaaggaggcg gcatggaaaa tcaagaacaa ccaggcaccg
     1141 acgccgtgga atgccccatg tgtggaggaa cgggcggttg gccaggcgta agcggctggg
     1201 ttgtctgccg gccctgcaat ggcactggaa cccccaagcc cgaggaatcg gcgtgagcgg
     1261 tcgcaaacca tccggcccgg tacaaatcgg cgcggcgctg ggtgatgacc tggtggagaa
     1321 gttgaaggcc gcgcaggccg cccagcggca acgcatcgag gcagaagcac gccccggtga
     1381 atcgtggcaa gcggccgctg atcgaatccg caaagaatcc cggcaaccgc cggcagccgg
     1441 tgcgccgtcg attaggaagc cgcccaaggg cgacgagcaa ccagattttt tcgttccgat
     1501 gctctatgac gtgggcaccc gcgatagtcg cagcatcatg gacgtggccg ttttccgtct
     1561 gtcgaagcgt gaccgacgag ctggcgaggt gatccgctac gagcttccag acgggcacgt
     1621 agaggtttcc gcagggccgg ccggcatggc cagtgtgtgg gattacgacc tggtactgat
     1681 ggcggtttcc catctaaccg aatccatgaa ccgataccgg gaagggaagg gagacaagcc
     1741 cggccgcgtg ttccgtccac acgttgcgga cgtactcaag ttctgccggc gagccgatgg
     1801 cggaaagcag aaagacgacc tggtagaaac ctgcattcgg ttaaacacca cgcacgttgc
     1861 catgcagcgt acgaagaagg ccaagaacgg ccgcctggtg acggtatccg agggtgaagc
     1921 cttgattagc cgctacaaga tcgtaaagag cgaaaccggg cggccggagt acatcgagat
     1981 cgagctagct gattggatgt accgcgagat cacagaaggc aagaacccgg acgtgctgac
     2041 ggttcacccc gattactttt tgatcgatcc cggcatcggc cgttttctct accgcctggc
     2101 acgccgcgcc gcaggcaagg cagaagccag atggttgttc aagacgatct acgaacgcag
     2161 tggcagcgcc ggagagttca agaagttctg tttcaccgtg cgcaagctga tcgggtcaaa
     2221 tgacctgccg gagtacgatt tgaaggagga ggcggggcag gctggcccga tcctagtcat
     2281 gcgctaccgc aacctgatcg agggcgaagc atccgccggt tcctaatgta cggagcagat
     2341 gctagggcaa attgccctag caggggaaaa aggtcgaaaa ggtctctttc ctgtggatag
     2401 cacgtacatt gggaacccaa agccgtacat tgggaaccgg aacccgtaca ttgggaaccc
     2461 aaagccgtac attgggaacc ggtcacacat gtaagtgact gatataaaag agaaaaaagg
     2521 cgatttttcc gcctaaaact ctttaaaact tattaaaact cttaaaaccc gcctggcctg
     2581 tgcataactg tctggccagc gcacagccga agagctgcaa aaagcgccta cccttcggtc
     2641 gctgcgctcc ctacgccccg ccgcttcgcg tcggcctatc gcggccgctg gccgctcaaa
     2701 aatggctggc ctacggccag gcaatctacc agggcgcgga caagccgcgc cgtcgccact
     2761 cgaccgccgg cgcccacatc aaggcaccgc gtgcctcact gattaagcat tggtaactgt
     2821 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     2881 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     2941 cgttccactg agcgtcagac cccttaataa gatgatcttc ttgagatcgt tttggtctgc
     3001 gcgtaatctc ttgctctgaa aacgaaaaaa ccgccttgca gggcggtttt tcgaaggttc
     3061 tctgagctac caactctttg aaccgaggta actggcttgg aggagcgcag tcaccaaaac
     3121 ttgtcctttc agtttagcct taaccggcgc atgacttcaa gactaactcc tctaaatcaa
     3181 ttaccagtgg ctgctgccag tggtgctttt gcatgtcttt ccgggttgga ctcaagacga
     3241 tagttaccgg ataaggcgca gcggtcggac tgaacggggg gttcgtgcat acagtccagc
     3301 ttggagcgaa ctgcctaccc ggaactgagt gtcaggcgtg gaatgagaca aacgcggcca
     3361 taacagcgga atgacaccgg taaaccgaaa ggcaggaaca ggagagcgca cgagggagcc
     3421 gccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc actgatttga
     3481 gcgtcagatt tcgtgatgct tgtcaggggg gcggagccta tggaaaaacg gctttgccgc
     3541 ggccctctca cttccctgtt aagtatcttc ctggcatctt ccaggaaatc tccgccccgt
     3601 tcgtaagcca tttccgctcg ccgcagtcga acgaccgagc gtagcgagtc agtgagcgag
     3661 gaagcggaat atatcctgta tcacatattc tgctgacgca ccggtgcagc cttttttctc
     3721 ctgccacatg aagcacttca ctgacaccct catcagtgcc aacatagtaa gccagtatac
     3781 actccgctag cgctgaggtc tgcctcgtga agaaggtgtt gctgactcat accaggctag
     3841 agcggccgcc acggcgatat cggatccata tgacgtcgac gcgtctgcag aagcttcgaa
     3901 ttcgagctcc cgggtaccat ggcatgcatc gatagatctc gaggcctgaa tcgccccatc
     3961 atccagccag aaagtgaggg agccacggtt gatgagagct ttgttgtagg tggaccagtt
     4021 ggtgattttg aacttttgct ttgccacgga acggtctgcg ttgtcgggaa gatgcgtgat
     4081 ctgatccttc aactcagcaa aagttcgatt tattcaacaa agccacgttg tgtctcaaaa
     4141 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
     4201 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcttgctc
     4261 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     4321 taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg atgcgccaga
     4381 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     4441 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     4501 tgatgatgca tggttactca ccactgcgat ccccgggaaa acagcattcc aggtattaga
     4561 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     4621 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     4681 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     4741 tggctggcct gttgaacaag tctggaaaga aatgcataag cttttgccat tctcaccgga
     4801 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     4861 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     4921 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     4981 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     5041 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     5101 cggcggcttt gttgaataaa tcgaactttt gctgagttga aggatcagat cacgcatctt
     5161 cccgacaacg cagaccgttc cgtggcaaag caaaagttca aaatcaccaa ctggtccacc
     5221 tacaacaaag ctctcatcaa ccgtggctcc ctcactttct ggctggatga tggggcgatt
     5281 caggcctcgg actagcatat gga
//
