pGreen 0029 vector (V010850)

Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V010850	pGreen 0029	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 pGreen0029 is a compact Agrobacterium binary vector with kanamycin-resistance genes for bacterial and plant transformation.

Vector Name:: pGreen 0029

Antibiotic Resistance:: Kanamycin

Length:: 4632 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 Vector Map

View Map in NovoBuilder

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.

pGreen 0029 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_22578        4632 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Compact Agrobacterium binary vector with a kanamycin-resistance 
            genes for bacterial and plant transformation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4632)
  AUTHORS   Hellens RP, Edwards EA, Leyland NR, Bean S, Mullineaux PM.
  TITLE     pGreen: a versatile and flexible binary Ti vector for 
            Agrobacterium-mediated plant transformation.
  JOURNAL   Plant Mol. Biol. 2000;42:819-32.
  PUBMED    10890530
REFERENCE   2  (bases 1 to 4632)
  AUTHORS   pGreen website
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 4632)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant Mol. 
            Biol."; date: "2000"; volume: "42"; pages: "819-32"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     This plasmid must be transformed into an Agrobacterium strain that 
            also carries a vector such as pSoup.
FEATURES             Location/Qualifiers
     source          1..4632
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(555..734)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     981..997
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1004..1022
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    complement(1031..1138)
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(1151..1169)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1190..1206)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1214..1230)
                     /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)."
     promoter        complement(1238..1269)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1284..1305)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    1544..1568
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(1659..2247)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2351..3163)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYGLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      3454..3889
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    4022..4044
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA
                     (truncated)"
     terminator      complement(4068..4319)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(join(4362..4632,1..521))
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERGRTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTQGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"