LOCUS       40924_33267        8253 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pNIC6012 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8253)
  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 8253)
  AUTHORS   Itoh Y, Nishijyo T.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-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:81-298-38-7997)
REFERENCE   3  (bases 1 to 8253)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8253)
  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 
            (23-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: "81-298-38-7997"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8253
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             195..881
                     /codon_start=1
                     /product="pVS1 resolvase"
                     /label=pVS1 resolvase
                     /note="not required for stable maintenance; orf1"
                     /protein_id="BAA96323.1"
                     /translation="MNKSAAAGLLGYARVSTDDQDLTNQRAELHAAGCTKLFSEKITGT
                     RRDRPELARMLDHLRPGDVVTVTRLDRLARSTRDLLDIAERIQEAGAGLRSLAEPWADT
                     TTPAGRMVLTVFAGIAEFERSLIIDRTRSGREAAKARGVKFGPRPTLTPAQIAHARELI
                     DQEGRTVKEAAALLGVHRSTLYRALERSEEVTPTEARRRGAFREDALTEADALAAAENE
                     RQEEQA"
     CDS             878..1093
                     /codon_start=1
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="not required for stable maintenance; orf2"
                     /protein_id="BAA96324.1"
                     /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 maintenance; orf3"
                     /protein_id="BAA96326.1"
                     /translation="MSKSTNTLSAGRPSARSSKAATLASLADTPAMKRVNFQLPAEDHT
                     KLKMYAVRQGKTITELLSEYIAQLPE"
     misc_feature    2076..2088
                     /label=KroB box
                     /note="KroB 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=multi cloning site
                     /note="multi cloning site"
     CDS             complement(6673..7530)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDKLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7531..7635)
                     /label=AmpR promoter
     oriT            8128..8237
                     /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 ttatgcattt ctttccagac
     5521 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
     5581 attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt
     5641 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
     5701 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt
     5761 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     5821 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
     5881 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc
     5941 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     6001 ggaatttaat cgcggcctcg agcaagacgt ttcccgttga atatggctca taacacccct
     6061 tgtattactg tttatgtaag cagacagttt tattgttcat gatgatatat ttttatcttg
     6121 tgcaatgtaa catcagagat tttgagacac aacgtggctt tgttgaataa atcgaacttt
     6181 tgctgagttg aaggatcaga tcacgcatct tcccgacaac gcagaccgtt ccgtggcaaa
     6241 gcaaaagttc aaaatcacca actggtccac ctacaacaaa gctctcatca accgtggctc
     6301 cctcactttc tggctggatg atggggcgat tcaggcctgg tatgagtcag caacaccttc
     6361 ttcacgaggc agacctcagc gcccgcgagc gagccgccga gctggccggc cagctcaccg
     6421 cacaccaggg acaggcggcc gcagttgctg gcacgcctgg caccggccga aaccaaaaca
     6481 cccagggcga agaagtaggg cggtggattg gtctggcggt agtggggtct gacgctcagt
     6541 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6601 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactg
     6661 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6721 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6781 atctggcccc agtgctgcaa tgataccgcg agatccacgc tcaccggctc cagatttatc
     6841 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6901 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6961 tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat
     7021 ggcttcattc agctccggtt cccaacgatc aaggcgggtt acatgatccc ccatgttgtg
     7081 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7141 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7201 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7261 accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcaccacata gcagaacttt
     7321 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7381 gttgagatcc agctcgatgt aacccactcg tgcacccaac ttatcttcag catcttttac
     7441 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7501 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7561 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7621 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     7681 tatcatgaca ttaacctata aaaataagcg tatcacgagg ccctttcgtc ttcaagaatt
     7741 ttataaaccg tggagcgggc aatactgagc tgatgagcaa tttccgttgc accagtgccc
     7801 ttctgatgaa gcgtcagcac gacgttcctg tccacggtac gcctgcggcc aaatttgatt
     7861 cctttcagct ttgcttcctg tcggccctca ttcgtgcgct ctaggatctc gcctgtcccc
     7921 tcagttcagt aatttcctgc atttgcctgt ttccagtcgg tagatattcc acaaaacagc
     7981 agggaagcag cgcttttccg ctgcataacc ctgcttcggg gtcattatag cgattttttc
     8041 ggtatatcca tcctttttcg cacgatatac aggattttgc caaagggttc gtgtagactt
     8101 tccttggtgt atccaacggc gtcagccggg caggataggt gaagtaggcc cacccgcgag
     8161 cgggtgttcc ttcttcactg tcccttattc gcacctggcg gtgctcaacg ggaatcctgc
     8221 tctgcgaggc tggccggcta ccgccggcgt aac
//
