Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V010892 | pCAMBIA1302 | 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 pCAMBIA1302 is a Agrobacterium binary vector for plant transformation, with hygromycin- and kanamycin-resistance and GFP genes.
- Vector Name:
- pCAMBIA1302
- Antibiotic Resistance:
- Kanamycin
- Length:
- 10549 bp
- Type:
- Plant Vectors
- Replication origin:
- ori
- Host:
- Plants
- Source/Author:
- Cambia
- Copy Number:
- High copy number
- Promoter:
- CaMV 35S (enhanced)
- 5' Primer:
- M13 fwd
- 3' Primer:
- M13 rev
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pCAMBIA1302 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
- Lim HS, Ko TS, Lambert KN, Kim HG, Korban SS, Hartman GL, Domier LL. Soybean mosaic virus helper component-protease enhances somatic embryo production and stabilizes transgene expression in soybean. Plant Physiol Biochem. 2005 Oct-Nov;43(10-11):1014-21. doi: 10.1016/j.plaphy.2005.08.012. Epub 2005 Oct 19. PMID: 16316753.
- Rizwan HM, Yang Q, Yousef AF, Zhang X, Sharif Y, Kaijie J, Shi M, Li H, Munir N, Yang X, Wei X, Oelmüller R, Cheng C, Chen F. Establishment of a Novel and Efficient Agrobacterium-Mediated in Planta Transformation System for Passion Fruit (Passiflora edulis). Plants (Basel). 2021 Nov 15;10(11):2459. doi: 10.3390/plants10112459. PMID: 34834821; PMCID: PMC8621743.
- Gupta T, Singhal T, Dhir S, Chandel V, Rishi N, Raj SK, Srivastava A. Demonstration of seed inoculation of dimer infectious clones of mungbean yellow mosaic India virus-soybean isolate. 3 Biotech. 2023 Nov;13(11):360. doi: 10.1007/s13205-023-03778-7. Epub 2023 Oct 11. PMID: 37840874; PMCID: PMC10567615.
pCAMBIA1302 vector Sequence
LOCUS Exported 10549 bp DNA circular SYN 03-SEP-2024
DEFINITION Agrobacterium binary vector for plant transformation, with
hygromycin- and kanamycin-resistance and GFP genes.
ACCESSION AF234298
VERSION .
KEYWORDS pCAMBIA1302
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10549)
AUTHORS Cambia
TITLE Direct Submission
REFERENCE 2 (bases 1 to 10549)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT The GenBank record was corrected by inserting a G at position 3443.
FEATURES Location/Qualifiers
source 1..10549
/lab_host="Plant Cells"
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 17..727
/codon_start=1
/gene="mgfp5"
/product="GFP with folding enhancement mutations"
/label=mgfp5
/note="mgfp5"
/note="suitable for expression in plants due to removal of
a cryptic intron"
/translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
ICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKAN
FKTRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
VTAAGITHGMDELYK"
CDS 734..751
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis affinity tag
/note="6xHis"
/translation="HHHHHH"
terminator 783..1035
/note="NOS terminator"
/note="nopaline synthase terminator and poly(A) signal"
misc_feature 1057..1081
/label=RB T-DNA repeat
/note="RB T-DNA repeat"
/note="right border repeat from nopaline C58 T-DNA"
CDS 2382..3011
/codon_start=1
/product="stability protein from Pseudomonas plasmid pVS1"
/label=stability protein from Pseudomonas plasmid pVS1
/note="pVS1 StaA"
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
CDS 3440..4513
/codon_start=1
/product="replication protein from Pseudomonas plasmid
pVS1"
/label=replication protein from Pseudomonas plasmid
pVS1
/note="pVS1 RepA"
/translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
LVMRYRNLIEGEASAGS"
rep_origin 4579..4773
/label=pVS1 oriV
/note="pVS1 oriV"
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
misc_feature 5117..5257
/label=bom
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(5443..6031)
/direction=LEFT
/label=ori
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6118..6912)
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/label=aphA-3
/note="KanR"
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
misc_feature 7337..7361
/label=LB T-DNA repeat
/note="LB T-DNA repeat"
/note="left border repeat from nopaline C58 T-DNA"
polyA_signal 7439..7613
/label=CaMV poly(A) signal
/note="CaMV poly(A) signal"
/note="cauliflower mosaic virus polyadenylation signal"
CDS complement(7654..8679)
/codon_start=1
/product="hygromycin B phosphotransferase"
/label=hygromycin B phosphotransferase
/note="HygR"
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
K"
promoter complement(8746..9422)
/note="CaMV 35S promoter (enhanced)"
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
promoter 9649..9679
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 9687..9703
/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 9711..9727
/label=M13 rev
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
CDS 9723..9955
/codon_start=1
/gene="lacZ (truncated)"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ (truncated)
/note="lacZ-alpha"
/translation="MT*LRIRARYPGIL*SRPAGMQAWHWPSFYNVVTGKTLALPNLIA
LQHIPLSPAGVIAKRPAPIALPNSCAA*MANA"
misc_feature 9736..9792
/label=MCS
/note="MCS"
/note="pUC18/19 multiple cloning site"
primer_bind complement(9796..9812)
/label=M13 fwd
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
promoter 10189..10534
/note="CaMV 35S promoter"
/note="strong constitutive promoter from cauliflower mosaic
virus"