LOCUS       40924_33307        8620 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pNIT6012 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8620)
  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 8620)
  AUTHORS   Itoh Y, Nishijyo T.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-MAY-2000) Yoshifumi Itoh, National Food Research 
            Institute, Division of Applied Microbiology; kannondai 2-1-2, 
            Tsukuba, Ibaraki 305-8642, Japan (E-mail:yosifumi@nfri.affrc.go.jp, 
            Tel:81-298-38-8075, Fax:81-298-38-7996)
REFERENCE   3  (bases 1 to 8620)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8620)
  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 
            (22-MAY-2000) Yoshifumi Itoh, National Food Research Institute, 
            Division of Applied Microbiology"; volume: " kannondai 2-1-2, 
            Tsukuba, Ibaraki 305-8642, Japan (E-mail:yosifumi@nfri.affrc.go.jp, 
            Tel:81-298-38-8075, Fax"; pages: "81-298-38-7996"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8620
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             195..881
                     /codon_start=1
                     /product="pVS1 resolvase"
                     /function="unknown"
                     /label=pVS1 resolvase
                     /note="not required for stable maintenance; orf1"
                     /protein_id="BAA96344.1"
                     /translation="MNKSAAAGLLGYARVSTDDQDLTNQRAELHAAGCTKLFSEKITGT
                     RRDRPELARMLDHLRPGDVVTVTRLDRLARSTRDLLDIAERIQEAGAGLRSLAEPWADT
                     TTPAGRMVLTVFAGIAEFERSLIIDRTRSGREAAKARGVKFGPRPTLTPAQIAHARELI
                     DQEGRTVKEAAALLGVHRSTLYRALERSEEVTPTEARRRGAFREDALTEADALAAAENE
                     RQEEQA"
     CDS             878..1093
                     /codon_start=1
                     /product="hypothetical protein"
                     /function="unknown"
                     /label=hypothetical protein
                     /note="not required for stable maintenance; orf2"
                     /protein_id="BAA96345.1"
                     /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"
                     /function="unknown"
                     /label=hypothetical protein
                     /note="not required for stable mainenance; orf3"
                     /protein_id="BAA96347.1"
                     /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
                     ALISRYKIVKSETGRPEYIEIELVDWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     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    5450..5502
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             complement(6292..6939)
                     /codon_start=1
                     /label=TetR
                     /note="tetracycline resistance regulatory protein"
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     CDS             7045..8241
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="VKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     oriT            8495..8604
                     /label=oriT
                     /note="incP origin of transfer"
ORIGIN
        1 agatccaacc 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 tagttgattg 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 accaggcctg aatcgcccca tcatccagcc
     5161 agaaagtgag ggagccacgg ttgatgagag ctttgttgta ggtggaccag ttggtgattt
     5221 tgaacttttg ctttgccacg gaacggtctg cgttgtcggg aagatgcgtg atctgatcct
     5281 tcaactcagc aaaagttcga tttattcaac aaagccacgt tgtgtctcaa aatctctgat
     5341 gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa
     5401 acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcttgc tcgagatcta
     5461 tcgatgcatg ccatggtacc cgggagctcg aattcgaagc ttttgccatt ctcaccggat
     5521 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
     5581 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
     5641 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
     5701 ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc
     5761 taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac ttgacgggac
     5821 ggcggctttg ttgaataaat cgaacttttg ctgagttgaa ggatcagatc acgcatcttc
     5881 ccgacaacgc agaccgttcc gtggcaaagc aaaagttcaa aatcaccaac tggtccacct
     5941 acaacaaagc tctcatcaac cgtggctccc tcactttctg gctggatgat ggggcgattc
     6001 aggcctggta tgagtcagca acaccttctt cacgaggcag acctcagcgc caggcgcacg
     6061 aggccggcgc gcggacccgc gccacggccc acgagcggac gccgcagcag gagcgccaga
     6121 aggccgccag agaggccgag cgcggccgtg aggcttggac gctagggcag ggcatgaaaa
     6181 agcccgtagc gggctgctac gggcgtctga cgcggtggaa agggggaggg gatgttgtct
     6241 acatggctct gctgtagtga gtgggttgcg ctccggcagc ggtcctgatc aatcgtcacc
     6301 ctttctcggt ccttcaacgt tcctgacaac gagcctcctt ttcgccaatc catcgacaat
     6361 caccgcgagt ccctgctcga acgctgcgtc cggaccggct tcgtcgaagg cgtctatcgc
     6421 ggcccgcaac agcggcgaga gcggagcctg ttcaacggtg ccgccgcgct cgccggcatc
     6481 gctgtcgccg gcctgctcct caagcacggc cccaacagtg aagtagctga ttgtcatcag
     6541 cgcattgacg gcgtccccgg ccgaaaaacc cgcctcgcag aggaagcgaa gctgcgcgtc
     6601 ggccgtttcc atctgcggtg cgcccggtcg cgtgccggca tggatgcgcg cgccatcgcg
     6661 gtaggcgagc agcgcctgcc tgaagctgcg ggcattcccg atcagaaatg agcgccagtc
     6721 gtcgtcggct ctcggcaccg aatgcgtatg attctccgcc agcatggctt cggccagtgc
     6781 gtcgagcagc gcccgcttgt tcctgaagtg ccagtaaagc gccggctgct gaacccccaa
     6841 ccgttccgcc agtttgcgtg tcgtcagacc gtctacgccg acctcgttca acaggtccag
     6901 ggcggcacgg atcactgtat tcggctgcaa ctttgtcatg cttgacactt tatcactgat
     6961 aaacataata tgtccaccaa cttatcagtg ataaagaatc cgcgcgttca atcggaccag
     7021 cggaggctgg tccggaggcc agacgtgaaa cccaacatac ccctgatcgt aattctgagc
     7081 actgtcgcgc tcgacgctgt cggcatcggc ctgattatgc cggtgctgcc gggcctcctg
     7141 cgcgatctgg ttcactcgaa cgacgtcacc gcccactatg gcattctgct ggcgctgtat
     7201 gcgttggtgc aatttgcctg cgcacctgtg ctgggcgcgc tgtcggatcg tttcgggcgg
     7261 cggccaatct tgctcgtctc gctggccggc gccactgtcg actacgccat catggcgaca
     7321 gcgcctttcc tttgggttct ctatatcggg cggatcgtgg ccggcatcac cggggcgact
     7381 ggggcggtag ccggcgctta tattgccgat atcactgatg gcgatgagcg cgcgcggcac
     7441 ttcggcttca tgagcgcctg tttcgggttc gggatggtcg cgggacctgt gctcggtggg
     7501 ctgatgggcg gtttctcccc ccacgctccg ttcttcgccg cggcagcctt gaacggcctc
     7561 aatttcctga cgggctgttt ccttttgccg gagtcgcaca aaggcgaacg ccggccgtta
     7621 cgccgggagg ctctcaaccc gctcgcttcg ttccggtggg cccggggcat gaccgtcgtc
     7681 gccgccctga tggcggtctt cttcatcatg caacttgtcg gacaggtgcc ggccgcgctt
     7741 tgggtcattt tcggcgagga tcgctttcac tgggacgcga ccacgatcgg catttcgctt
     7801 gccgcatttg gcattctgca ttcactcgcc caggcaatga tcaccggccc tgtagccgcc
     7861 cggctcggcg aaaggcgggc actcatgctc ggaatgattg ccgacggcac aggctacatc
     7921 ctgcttgcct tcgcgacacg gggatggatg gcgttcccga tcatggtcct gcttgcttcg
     7981 ggtggcatcg gaatgccggc gctgcaagca atgttgtcca ggcaggtgga tgaggaacgt
     8041 caggggcagc tgcaaggctc actggcggcg ctcaccagcc tgacctcgat cgtcggaccc
     8101 ctcctcttca cggcgatcta tgcggcttct ataacaacgt ggaacgggtg ggcatggatt
     8161 gcaggcgctg ccctctactt gctctgcctg ccggcgctgc gtcgcgggct ttggagcggc
     8221 gcagggcaac gagccgatcg ctgatcgtgg aaacgatagg gacgtcatat ggatctcgcc
     8281 tgtcccctca gttcagtaat ttcctgcatt tgcctgtttc cagtcggtag atattccaca
     8341 aaacagcagg gaagcagcgc ttttccgctg cataaccctg cttcggggtc attatagcga
     8401 ttttttcggt atatccatcc tttttcgcac gatatacagg attttgccaa agggttcgtg
     8461 tagactttcc ttggtgtatc caacggcgtc agccgggcag gataggtgaa gtaggcccac
     8521 ccgcgagcgg gtgttccttc ttcactgtcc cttattcgca cctggcggtg ctcaacggga
     8581 atcctgctct gcgaggctgg ccggctaccg ccggcgtaac
//
