Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V010906 | pBINPLUS | In stock, instant shipping |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pBINPLUS
- Antibiotic Resistance:
- Kanamycin
- Length:
- 12396 bp
- Type:
- Plant Vectors
- Replication origin:
- oriV
- Host:
- Plants
- Source/Author:
- van Engelen FA, Molthoff JW, Conner AJ, Nap JP, Pereira A, Stiekema
- Selection Marker:
- Neomycin/G418(Geneticin)
- Copy Number:
- High copy number
- Promoter:
- NOS
- 5' Primer:
- M13 fwd
- 3' Primer:
- M13 rev
- Growth Strain(s):
- stbl3
- Growth Temperature:
- 37℃
pBINPLUS 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.
pBINPLUS vector Sequence
LOCUS pBINPLUS. 12396 bp DNA circular SYN 01-JAN-1980
DEFINITION Improved binary Agrobacterium vector for plant transformation.
ACCESSION .
VERSION .
KEYWORDS pBINPLUS
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12396)
AUTHORS van Engelen FA, Molthoff JW, Conner AJ, Nap JP, Pereira A, Stiekema
WJ.
TITLE pBINPLUS: an improved plant transformation vector based on pBIN19.
JOURNAL Transgenic Res. 1995;4:288-90.
PUBMED 7655517
REFERENCE 2 (bases 1 to 12396)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 12396)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Transgenic
Res."; date: "1995"; volume: "4"; pages: "288-90"
COMMENT SGRef: number: 2; type: "Journal Article"
COMMENT This sequence was reconstructed using the original paper. The
orientation of the truncated pBR322 origin is uncertain.
FEATURES Location/Qualifiers
source 1..12396
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 6..616
/label=oriV
/note="origin of replication for the bacterial F plasmid"
rep_origin 1758..2110
/direction=RIGHT
/label=pBR322 ori (truncated)
/note="pBR322 ori (truncated)"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication (truncated)"
CDS 2871..3662
/label=KanR
/note="aminoglycoside phosphotransferase"
CDS 3964..5109
/label=trfA
/note="trans-acting replication protein that binds to and
activates oriV"
misc_feature 6708..6732
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
promoter 7205..7388
/label=NOS promoter
/note="nopaline synthase promoter"
CDS 7409..8200
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
terminator 8593..8845
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
protein_bind 9394..9415
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 9430..9460
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 9468..9484
/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 9492..9508
/label=M13 rev
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
misc_feature complement(9534..9590)
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(9605..9621)
/label=M13 fwd
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 9919..9943
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
rep_origin 10000..10624
/label=oriV
/note="incP origin of replication"
oriT 11739..11848
/label=oriT
/note="incP origin of transfer"
CDS 11881..12249
/label=traJ
/note="oriT-recognizing protein"