pNIT6011 vector (V004306)

Basic Vector Information

Vector Name:
pNIT6011
Antibiotic Resistance:
Tetracycline
Length:
8620 bp
Type:
Shuttle vector
Replication origin:
p15A ori
Source/Author:
Heeb S, Itoh Y, Nishijyo T, Schnider U, Keel C, Wade J, Walsh U, O'Gara F, Haas D.

pNIT6011 vector Map

pNIT60118620 bp4008001200160020002400280032003600400044004800520056006000640068007200760080008400pVS1 resolvasepVS1 StaAhypothetical proteinKorB boxpVS1 RepApVS1 oriVp15A orip15A origin of transfermultiple cloning siteTetRTcRoriT

pNIT6011 vector Sequence

LOCUS       40924_33302        8620 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pNIT6011 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="BAA96337.1"
                     /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="BAA96338.1"
                     /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="BAA96340.1"
                     /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
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPNVLTVHPDYFLIDPGIGRFLYR
                     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"

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