Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V008762 | pCAMBIA1304 | 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 pCAMBIA1304 is a agrobacterium binary vector for plant transformation, with hygromycin- and kanamycin-resistance and GFP-GUS genes.
- Vector Name:
- pCAMBIA1304
- Antibiotic Resistance:
- Kanamycin
- Length:
- 12361 bp
- Type:
- Binary vector
- Replication origin:
- ori
- Source/Author:
- Hajdukiewicz P, Svab Z, Maliga P.
- Promoter:
- CaMV35S(enhanced)
- Growth Strain(s):
- stbl3
- Growth Temperature:
- 37℃
pCAMBIA1304 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
- Rahimifard Hamedani P, Solouki M, Ehsani P, Emamjomeh A, Ofoghi H. Expression of BMP2-Hydrophobin fusion protein in the tobacco plant and molecular dynamic evaluation of its simulated model. Plant Biotechnol Rep. 2021;15(3):309-316.
pCAMBIA1304 vector Sequence
LOCUS 40924_8721 12361 bp DNA circular SYN 17-DEC-2018
DEFINITION Binary vector pCAMBIA-1304, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12361)
AUTHORS Hajdukiewicz P, Svab Z, Maliga P.
TITLE The small, versatile pPZP family of Agrobacterium binary vectors for
plant transformation
JOURNAL Plant Mol. Biol. 25 (6), 989-994 (1994)
PUBMED 7919218
REFERENCE 2 (bases 1 to 12361)
AUTHORS Roberts C, Rajagopal S, Smith LM, Nguyen TA, Yang W, Nugrohu S, Ravi
KS, Vijayachandra K, Harcourt RL, Dransfield L, Desamero N, Slamet
I, Hadjukiewicz P, Svab Z, Maliga P, Mayer JE, Keese PK, Kilian A,
Jefferson RA.
TITLE A comprehensive set of modular vectors for advanced manipulations
and efficient transformation of plants
JOURNAL Unpublished
REFERENCE 3 (bases 1 to 12361)
AUTHORS Roberts C, Rajagopal S, Smith LM, Nguyen TA, Yang W, Nugrohu S, Ravi
KS, Vijayachandra K, Harcourt RL, Dransfield L, Desamero N, Slamet
I, Hadjukiewicz P, Svab Z, Maliga P, Mayer JE, Keese PK, Kilian A,
Jefferson RA.
TITLE Direct Submission
JOURNAL Submitted (15-FEB-2000) CAMBIA, Clunies Ross St, Black Mountain /
GPO Box 3200, Canberra, ACT 2601, Australia
REFERENCE 4 (bases 1 to 12361)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 12361)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plant Mol.
Biol."; date: "1994"; volume: "25"; issue: "6"; pages: "989-994"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
(15-FEB-2000) CAMBIA, Clunies Ross St, Black Mountain / GPO Box
3200, Canberra, ACT 2601, Australia"
COMMENT SGRef: number: 4; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..12361
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 17..727
/codon_start=1
/label=mgfp5
/note="GFP with folding enhancement mutations"
/translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
ICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKAN
FKTRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
VTAAGITHGMDELYK"
CDS 734..2539
/codon_start=1
/label=GUS
/note="beta-glucuronidase"
/translation="LRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAIA
VPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEVM
EHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHDF
FNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQV
VATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGQQFL
INHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDWA
DEHGIVVIDETAAVGFQLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARDK
NHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISDL
FDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMYT
DMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRKP
KSAAFLLQKRWTGMNFGEKPQQGGKQ"
CDS 2546..2563
/codon_start=1
/label=6xHis
/note="6xHis affinity tag"
/translation="HHHHHH"
terminator 2595..2847
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
misc_feature 2869..2893
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
CDS 4193..4819
/codon_start=1
/label=pVS1 StaA
/note="stability protein from the Pseudomonas plasmid pVS1
(Heeb et al., 2000)"
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
CDS 5256..6320
/codon_start=1
/label=pVS1 RepA
/note="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
MRYRNLIEGEASAGS"
rep_origin 6389..6583
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
misc_feature 6927..7067
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(7253..7841)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7931..8722)
/codon_start=1
/label=KanR
/note="aminoglycoside phosphotransferase"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
misc_feature 9147..9171
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
polyA_signal complement(9249..9423)
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
CDS complement(9466..10488)
/codon_start=1
/label=HygR
/note="aminoglycoside phosphotransferase from E. coli"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
K"
promoter complement(10556..11233)
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
protein_bind 11424..11445
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 11460..11490
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 11498..11514
/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 11522..11538
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 11548..11604
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(11608..11624)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 12001..12346
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"