Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V010908 | pBI221 | 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:
- pBI221
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5667 bp
- Type:
- Plant Vectors
- Replication origin:
- ori
- Host:
- Plants
- Source/Author:
- Chen PY, Wang CK, Soong SC, To KY.
- Copy Number:
- High copy number
- Promoter:
- CaMV35S(long)
- 5' Primer:
- M13 fwd
- 3' Primer:
- M13 rev
pBI221 vector 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.
pBI221 vector Sequence
LOCUS 40924_6402 5667 bp DNA circular SYN 17-DEC-2018
DEFINITION Transient expression vector pBI221, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5667)
AUTHORS Chen PY, Wang CK, Soong SC, To KY.
TITLE Complete sequence of the binary vector pBI121 and its application in
cloning T-DNA insertion from transgenic plants
JOURNAL Mol. Breed. 11, 287-293 (2003)
REFERENCE 2 (bases 1 to 5667)
AUTHORS Chen PY, Wang CK, To KY.
TITLE Direct Submission
JOURNAL Submitted (12-APR-2002) Institute of Bioagricultural Sciences,
Academia Sinica, Taipei 11529, Taiwan
REFERENCE 3 (bases 1 to 5667)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 5667)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Mol. Breed.
11, 287-293 (2003)"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(12-APR-2002) Institute of Bioagricultural Sciences, Academia
Sinica, Taipei 11529, Taiwan"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5667
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 512..857
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
CDS 896..2704
/codon_start=1
/label=GUS
/note="beta-glucuronidase"
/translation="MLRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
terminator 2778..3030
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
primer_bind complement(3039..3055)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 3529..3633
/label=AmpR promoter
CDS 3634..4491
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 4665..5253
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 5541..5562
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5577..5607
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5615..5631
/label=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 5639..5655
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"