Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V010909 | pBI121 | 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 pBI121 is a binary Agrobacterium vector equipped with a GUS reporter gene designed for facilitating plant transformation.
- Vector Name:
- pBI121
- Antibiotic Resistance:
- Kanamycin
- Length:
- 14758 bp
- Type:
- Plant Vectors
- Replication origin:
- oriV
- Host:
- Plants
- Source/Author:
- Chen PY, Wang CK, Soong SC, To KY.
- Selection Marker:
- Neomycin/G418(Geneticin)
- Copy Number:
- High copy number
- Promoter:
- CaMV35S(long)
- 5' Primer:
- M13 fwd
- 3' Primer:
- M13 rev
- Growth Strain(s):
- stbl3
- Growth Temperature:
- 37℃
pBI121 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
- Srinivas K, Muralikrishna N, Kumar KB, Raghu E, Mahender A, Kiranmayee K, Yashodahara V, Sadanandam A. Biolistic transformation of Scoparia dulcis L. Physiol Mol Biol Plants. 2016 Jan;22(1):61-8. doi: 10.1007/s12298-016-0338-2. Epub 2016 Jan 20. PMID: 27186019; PMCID: PMC4840147.
- Beihaghi M, Marashi H, Bagheri A, Sankian M. Transient expression of CCL21as recombinant protein in tomato. Biotechnol Rep (Amst). 2017 Nov 26;17:10-15. doi: 10.1016/j.btre.2017.11.007. PMID: 29276695; PMCID: PMC5730375.
pBI121 vector Sequence
LOCUS Exported 14758 bp DNA circular SYN 03-SEP-2024
DEFINITION Binary vector pBI121, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14758)
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 14758)
AUTHORS To KY.
TITLE Direct Submission
JOURNAL Submitted (20-FEB-2002) Institute of BioAgricultural Sciences,
Academia Sinica, Taipei 11529, Taiwan
REFERENCE 3 (bases 1 to 14758)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 14758)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 14758)
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
(20-FEB-2002) Institute of BioAgricultural Sciences, Academia
Sinica, Taipei 11529, Taiwan"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT SGRef: number: 4; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..14758
/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"
rep_origin complement(790..1168)
/direction=LEFT
/note="ColE1 ori; similar to sequence in GenBank Accession
Number V00268"
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"
rep_origin complement(4135..4760)
/direction=LEFT
/label=oriV
/note="incP origin of replication"
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 operator
/bound_moiety="lac repressor encoded by lacI"
/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="common sequencing primer, one of multiple similar
variants"
promoter 5461..5806
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
CDS 5845..7653
/label=GUS
/note="beta-glucuronidase"
regulatory 7727..7979
/label=NOS
/note="NOS"
/regulatory_class="terminator"
terminator 7727..7979
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
primer_bind complement(7988..8004)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature complement(8622..8646)
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
misc_feature complement(9156..10198)
/label=similar to tetA in GenBank Accession Number L13842
/note="similar to tetA in GenBank Accession Number L13842"
CDS complement(10245..11390)
/label=trfA
/note="trans-acting replication protein that binds to and
activates oriV"
CDS complement(11692..12483)
/label=KanR
/note="aminoglycoside phosphotransferase"
mobile_element 12675..13442
/label=IS1
/note="prokaryotic transposable element"
misc_feature complement(13444..13794)
/note="similarity to NPT III gene in GenBank Accession
Number V01547"
misc_feature complement(13795..14066)
/label=similar to kilA in GenBank Accession Number M62846
/note="similar to kilA in GenBank Accession Number M62846"
rep_origin complement(14143..14753)
/direction=LEFT
/label=oriV
/note="origin of replication for the bacterial F plasmid"