LOCUS       Exported                8620 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pNIT6012 DNA, complete sequence.
ACCESSION   AB043476
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, 
            OGara 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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8620
                     /organism="Shuttle vector pNIT6012"
                     /lab_host="Escherichia coli, Pseudomonas aeruginosa"
                     /mol_type="other DNA"
                     /db_xref="taxon:126709"
     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..1809
                     /codon_start=1
                     /gene="staA"
                     /product="pVS1 partitioning protein StaA"
                     /label=staA
                     /note="similar to IncC2"
                     /experiment="experimental evidence, no additional details 
                     recorded"
                     /protein_id="BAA96346.1"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     gene            1180..1809
                     /gene="staA"
                     /label=staA
     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..3311
                     /codon_start=1
                     /gene="repA"
                     /product="pVS1 replication protein RepA"
                     /function="pVS1 replication protein"
                     /label=repA
                     /note="high copy mutant RepA"
                     /protein_id="BAA96348.1"
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELVDWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     gene            2238..3311
                     /gene="repA"
                     /label=repA
     misc_difference 2974
                     /gene="repA"
                     /replace="c"
                     /note="present in pME290, pME6010, pNIC6010 and pNIT6010; 
                     results in decreased copy number"
     rep_origin      3377..3571
                     /label=pVS1 origin of replication
                     /note="pVS1 origin of replication"
     misc_feature    3377..3384
                     /label=DnaA box
                     /note="DnaA box"
     repeat_region   3388..3471
                     /label=21 bp
                     /note="21 bp"
     misc_feature    3472..3563
                     /label=AT-rich region
                     /note="AT-rich region"
     misc_feature    3564..3571
                     /label=DnaA box
                     /note="DnaA box"
     rep_origin      4264..5093
                     /label=p15A origin of replication
                     /note="p15A origin of replication"
     rep_origin      4281..4826
                     /direction=RIGHT
                     /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(6289..6966)
                     /codon_start=1
                     /gene="tetR"
                     /product="RK2 tetracycline repressor protein TetR"
                     /label=tetR
                     /protein_id="BAA96349.1"
                     /translation="MFISDKVSSMTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGV
                     QQPALYWHFRNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRD
                     GARIHAGTRPGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSD
                     AGERGGTVEQAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRK
                     GDD"
     gene            complement(6289..6966)
                     /gene="tetR"
                     /label=tetR
     CDS             complement(6289..6939)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     CDS             6970..8244
                     /codon_start=1
                     /gene="tetA"
                     /product="RK2 tetracycline resistance protein TetA"
                     /label=tetA
                     /protein_id="BAA96350.1"
                     /translation="MSTNLSVIKNPRVQSDQRRLVRRPDVKPNIPLIVILSTVALDAVG
                     IGLIMPVLPGLLRDLVHSNDVTAHYGILLALYALVQFACAPVLGALSDRFGRRPILLVS
                     LAGATVDYAIMATAPFLWVLYIGRIVAGITGATGAVAGAYIADITDGDERARHFGFMSA
                     CFGFGMVAGPVLGGLMGGFSPHAPFFAAAALNGLNFLTGCFLLPESHKGERRPLRREAL
                     NPLASFRWARGMTVVAALMAVFFIMQLVGQVPAALWVIFGEDRFHWDATTIGISLAAFG
                     ILHSLAQAMITGPVAARLGERRALMLGMIADGTGYILLAFATRGWMAFPIMVLLASGGI
                     GMPALQAMLSRQVDEERQGQLQGSLAALTSLTSIVGPLLFTAIYAASITTWNGWAWIAG
                     AALYLLCLPALRRGLWSGAGQRADR"
     gene            6970..8244
                     /gene="tetA"
                     /label=tetA
     CDS             7045..8244
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     misc_feature    8247..8620
                     /label=RP4 mob region
                     /note="RP4 mob region"
     oriT            8495..8604
                     /note="incP origin of transfer"