Basic Vector Information
- Vector Name:
- pNIC6012
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8253 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.
pNIC6012 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 ℃.
pNIC6012 vector Sequence
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"
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