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 Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
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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