Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V010850 | pGreen 0029 | In stock, instant shipping |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
The pGreen0029 is a compact Agrobacterium binary vector with kanamycin-resistance genes for bacterial and plant transformation.
- Vector Name:
- pGreen 0029
- Antibiotic Resistance:
- Kanamycin
- Length:
- 4629 bp
- Type:
- Plant Vectors
- Replication origin:
- pSa ori
- Host:
- Plants
- Source/Author:
- Hellens RP, Edwards EA, Leyland NR, Bean S, Mullineaux PM.
- Selection Marker:
- Neomycin/G418(Geneticin)
- Copy Number:
- High copy number
- Promoter:
- NOS
- 5' Primer:
- M13 fwd
- 3' Primer:
- M13 rev
pGreen 0029 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
- Bukhari SA, Mustafa G, Bashir S, et al. Genetic transformation of Sr22 gene in a high yielding susceptible cultivar of commercial wheat (Triticum aestivum L.). 3 Biotech. 2020;10(5):197. doi:10.1007/s13205-020-02185-6
pGreen 0029 vector Sequence
LOCUS Exported 4629 bp DNA circular SYN 21-JUL-2025
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4629)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..4629
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 95..116
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 131..161
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 169..185
/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)."
CDS 205..570
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ-alpha
/translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
TDRPSQQLRSLNGEWKL"
promoter 230..248
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
misc_feature 261..368
/label=MCS
/note="pBluescript multiple cloning site"
primer_bind 285..301
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(335..351)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(377..395)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 668..847
/label=NOS promoter
/note="nopaline synthase promoter"
CDS 881..1675
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
terminator 1715..1967
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
misc_feature 1991..2013
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA
(truncated)"
rep_origin complement(2080..2668)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2769..3584)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
rep_origin 3875..4310
/label=pSa ori
/note="origin of replication from bacterial plasmid pSa"
misc_feature 4461..4485
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"