Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V005425 | pGWB17 | In stock, 1 week for quality controls |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pGWB17
- Antibiotic Resistance:
- Kanamycin
- Length:
- 17407 bp
- Type:
- Plant binary expression vectors, Gateway vectors
- Replication origin:
- oriV
- Host:
- Plants
- Selection Marker:
- Neomycin, Hygromycin
- Promoter:
- CaMV35S(long)
- Fusion Tag:
- Myc
pGWB17 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.
pGWB17 vector Sequence
LOCUS 40924_22829 17407 bp DNA circular SYN 13-JAN-2022
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 17407)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 17407)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..17407
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(148..516)
/label=traJ
/note="oriT-recognizing protein"
oriT complement(549..658)
/direction=LEFT
/label=oriT
/note="incP origin of transfer"
CDS complement(1342..1989)
/label=TetR
/note="tetracycline resistance regulatory protein"
misc_feature complement(2454..2478)
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
promoter 2634..2817
/label=NOS promoter
/note="nopaline synthase promoter"
CDS 2841..3629
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase from Tn5"
terminator 4022..4274
/note="NOS terminator"
/note="nopaline synthase terminator and poly(A) signal"
protein_bind 4823..4844
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4859..4889
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4897..4913
/label=lac repressor encoded by lacI binding site
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 4921..4937
/label=M13 rev
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
promoter 5461..5806
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
protein_bind 5828..5952
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter 5989..6019
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 6073..6729
/label=CmR
/note="chloramphenicol acetyltransferase"
rep_origin complement(6784..7408)
/direction=LEFT
/label=oriV
/note="incP origin of replication"
protein_bind complement(7420..7544)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
CDS 7570..7599
/label=Myc
/note="Myc (human c-Myc proto-oncogene) epitope tag"
CDS 7606..7635
/codon_start=1
/product="Myc (human c-Myc oncogene) epitope tag"
/label=Myc
/note="Myc"
/translation="EQKLISEEDL"
CDS 7651..7680
/codon_start=1
/product="Myc (human c-Myc oncogene) epitope tag"
/label=Myc
/note="Myc"
/translation="EQKLISEEDL"
CDS 7687..7716
/codon_start=1
/product="Myc (human c-Myc oncogene) epitope tag"
/label=Myc
/note="Myc"
/translation="EQKLISEEDL"
terminator 7747..7999
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
terminator complement(8428..8680)
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
CDS complement(9151..10173)
/label=HygR
/note="aminoglycoside phosphotransferase from E. coli"
promoter complement(10183..10527)
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
primer_bind complement(10637..10653)
/label=M13 fwd
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
misc_feature complement(11271..11295)
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
CDS complement(12894..14039)
/label=trfA
/note="trans-acting replication protein that binds to and
activates oriV"
CDS complement(14341..15132)
/label=KanR
/note="aminoglycoside phosphotransferase"
mobile_element 15324..16091
/label=IS1
/note="prokaryotic transposable element"
rep_origin complement(16792..17402)
/direction=LEFT
/label=oriV
/note="origin of replication for the bacterial F plasmid"