LOCUS       Exported                8253 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pNIC6010 DNA, complete sequence.
ACCESSION   AB043580
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, 
            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 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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8253
                     /organism="Shuttle vector pNIC6010"
                     /lab_host="Escherichia coli, Pseudomonas aeruginosa"
                     /mol_type="other DNA"
                     /db_xref="taxon:126840"
     CDS             195..881
                     /codon_start=1
                     /product="pVS1 resolvase"
                     /label=pVS1 resolvase
                     /note="not required for stable maintenance; orf1"
                     /protein_id="BAA96311.1"
                     /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="BAA96312.1"
                     /translation="MKPHQDGQDEPFFITEEIEAEMIAAGYVFEPPAHVSTVRLHEILA
                     GLSDAKLAAWPASLAAEETERRRLKR"
     CDS             1180..1809
                     /codon_start=1
                     /gene="staA"
                     /product="pVS1 partitioning protein StaA"
                     /label=staA
                     /note="similar to IncC2; stable maintenance of pVS1 
                     replicon"
                     /protein_id="BAA96313.1"
                     /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 maintenance; orf3"
                     /protein_id="BAA96314.1"
                     /translation="MSKSTNTLSAGRPSARSSKAATLASLADTPAMKRVNFQLPAEDHT
                     KLKMYAVRQGKTITELLSEYIAQLPE"
     misc_feature    2076..2088
                     /label=KroB box
                     /note="KroB box"
     CDS             2238..3311
                     /codon_start=1
                     /gene="repA"
                     /product="pVS1 replication protein RepA"
                     /label=repA
                     /protein_id="BAA96315.1"
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     gene            2238..3311
                     /gene="repA"
                     /label=repA
     misc_difference 2974
                     /gene="repA"
                     /replace="t"
                     /note="resent in pME294, pME6012, pNIC6012 and pNIT6012; 
                     results in increased copy number"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     misc_difference 3015
                     /gene="repA"
                     /replace="a"
                     /note="present in pME292, pME6011, pNIC6011 and pNIT6011; 
                     results in decreased copy number"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     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 repeats
                     /note="21 bp repeats"
     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=multi cloning site
                     /note="multi cloning site"
     CDS             complement(6670..7530)
                     /codon_start=1
                     /gene="bla"
                     /product="beta lactamase"
                     /function="ampicillin/carbenycillin resistance"
                     /label=bla
                     /note="derived from Tn801"
                     /protein_id="BAA96316.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDKLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(6670..7530)
                     /gene="bla"
                     /label=bla
     promoter        complement(7531..7635)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7907..8253
                     /label=mob region from RP4
                     /note="mob region from RP4"
     oriT            8128..8237
                     /note="incP origin of transfer"