Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V004776 | pME6031 | In stock, instant shipping |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
The pME6031 is a broad-host vector for studies requiring multi-color tagging and visualization of plant-associated, Gram-negative bacterial strains.
- Vector Name:
- pME6031
- Antibiotic Resistance:
- Tetracycline
- Length:
- 8310 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.
- Growth Strain(s):
- DH5a
- Growth Temperature:
- 37℃
pME6031 vector Map
Plasmid 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 ℃.
6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it
General Plasmid Transform Protocol
1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.
2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.
3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.
4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.
5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.
References
- Wilton R, Ahrendt AJ, Shinde S, et al. A New Suite of Plasmid Vectors for Fluorescence-Based Imaging of Root Colonizing Pseudomonads. Front Plant Sci. 2018;8:2242. Published 2018 Feb 1. doi:10.3389/fpls.2017.02242
pME6031 vector Sequence
LOCUS 40924_30555 8310 bp DNA circular SYN 18-DEC-2018
DEFINITION Shuttle vector pME6031, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8310)
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 8310)
AUTHORS Heeb S.
TITLE Direct Submission
JOURNAL Submitted (30-DEC-1998) Laboratoire de Biologie Microbienne,
University of Lausanne, Batiment de Biologie, Lausanne CH-1015,
Switzerland
REFERENCE 3 (bases 1 to 8310)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 8310)
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
(30-DEC-1998) Laboratoire de Biologie Microbienne, University of
Lausanne, Batiment de Biologie, Lausanne CH-1015, Switzerland"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8310
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 195..881
/codon_start=1
/product="pVS1 resolvase"
/label=pVS1 resolvase
/note="ORF1; not required for stable maintenance"
/protein_id="AAD19683.1"
/translation="MNKSAAAGLLGYARVSTDDQDLTNQRAELHAAGCTKLFSEKITGT
RRDRPELARMLDHLRPGDVVTVTRLDRLARSTRDLLDIAERIQEAGAGLRSLAEPWADT
TTPAGRMVLTVFAGIAEFERSLIIDRTRSGREAAKARGVKFGPRPTLTPAQIAHARELI
DQEGRTVKEAAALLGVHRSTLYRALERSEEVTPTEARRRGAFREDALTEADALAAAENE
RQEEQA"
CDS 878..1093
/codon_start=1
/product="hypothetical protein"
/label=hypothetical protein
/note="ORF2; not required for stable maintenance"
/protein_id="AAD19684.1"
/translation="MKPHQDGQDEPFFITEEIEAEMIAAGYVFEPPAHVSTVRLHEILA
GLSDAKLAAWPASLAAEETERRRLKR"
CDS 1180..1806
/label=pVS1 StaA
/note="stability protein from the Pseudomonas plasmid pVS1
(Heeb et al., 2000)"
CDS 1830..2045
/codon_start=1
/product="hypothetical protein"
/label=hypothetical protein
/note="ORF3; not required for stable maintenance"
/protein_id="AAD19686.1"
/translation="MSKSTNTLSAGRPSARSSKAATLASLADTPAMKRVNFQLPAEDHT
KLKMYAVRQGKTITELLSEYIAQLPE"
misc_feature 2075..2088
/label=KorB box
/note="KorB box"
CDS 2238..3308
/label=pVS1 RepA
/note="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
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_difference 5136..5414
/label=part of terminator absent in pME6030
/note="part of terminator absent in pME6030"
regulatory 5148..5414
/label=T4 transcription terminator
/note="T4 transcription terminator"
/regulatory_class="terminator"
misc_feature 5416..5526
/label=multiple cloning site
/note="multiple cloning site"
CDS complement(6318..6965)
/label=TetR
/note="tetracycline resistance regulatory protein"
CDS 7071..8267
/label=TcR
/note="tetracycline efflux protein"