Price Information
Cat No. | Plasmid Name | Availability | Add to cart |
---|---|---|---|
V007258 | pART27 | 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
pART27 is a plant expression vector.
- Vector Name:
- pART27
- Antibiotic Resistance:
- Spectinomycin
- Length:
- 11612 bp
- Type:
- Plant Expression Vectors
- Replication origin:
- oriV
- Host:
- Plants
- Selection Marker:
- nptII
- Promoter:
- NOS
pART27 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
- Benny A, Alex S, Soni KB, Anith KN, Kiran AG, Viji MM. Improved transformation of Agrobacterium assisted by silver nanoparticles. BioTechnologia (Pozn). 2022 Sep 29;103(3):311-317.
pART27 vector Sequence
LOCUS Exported 11612 bp DNA circular SYN 02-SEP-2024 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11612) TITLE Direct Submission REFERENCE 2 (bases 1 to 11612) TITLE Direct Submission REFERENCE 3 (bases 1 to 11612) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" COMMENT SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..11612 /mol_type="other DNA" /organism="synthetic DNA construct" misc_feature complement(480..504) /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" protein_bind 759..780 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 795..825 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 833..849 /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 857..873 /label=M13 rev /note="M13 rev" /note="common sequencing primer, one of multiple similar variants" promoter 891..909 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" promoter complement(1029..1047) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1054..1070) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 1228..1411 /label=NOS promoter /note="nopaline synthase promoter" CDS 1412..2230 /label=KanR /note="fusion between an N-terminal peptide of nopaline synthase and Tn5 aminoglycoside phosphotransferase" terminator 2858..3110 /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" misc_feature complement(3173..3197) /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" oriT 3752..3861 /label=oriT /note="incP origin of transfer" mobile_element 3921..4688 /label=IS1 /note="prokaryotic transposable element" CDS 4688..5038 /label=traJ /note="oriT-recognizing protein" CDS 5313..6458 /label=trfA /note="trans-acting replication protein that binds to and activates oriV" rep_origin complement(7821..8409) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS 9506..10294 /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=aadA /note="SmR" /note="confers resistance to spectinomycin and streptomycin" /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEDRLASRADQLEEFVHYVKGEITKVVGK" rep_origin complement(10899..11612) /direction=LEFT /label=oriV /note="incP origin of replication"