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pGreenII 0579 vector (V010845)

Price Information

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V010845 pGreenII 0579 In stock, instant shipping

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Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

The pGreenII 0579 is a agrobacterium binary vector with kanamycin- resistance and luciferase genes.

Vector Name:
pGreenII 0579
Antibiotic Resistance:
Kanamycin
Length:
5663 bp
Type:
Plant Vectors
Replication origin:
pSa ori
Host:
Plants
Source/Author:
Hellens RP, Edwards EA, Leyland NR, Bean S, Mullineaux PM.
Copy Number:
High copy number
Promoter:
CaMV 35S
5' Primer:
M13 fwd
3' Primer:
M13 rev
Growth Strain(s):
Top10
Growth Temperature:
37℃

pGreenII 0579 vector Map

Copy Sequence View Map
pGreenII 05795663 bp60012001800240030003600420048005400pSa oriLB T-DNA repeatCaMV poly(A) signalluciferaseCaMV 35S promoterM13 fwdT7 promoterMCST3 promoterM13 revlac operatorlac promoterCAP binding siteRB T-DNA repeatoriKanR

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.

pGreenII 0579 vector Sequence

LOCUS       pGreenII_0579.        5663 bp DNA     circular SYN 06-JAN-2024
DEFINITION  Agrobacterium binary vector with kanamycin- resistance and 
            luciferase genes.
ACCESSION   .
VERSION     .
KEYWORDS    pGreenII 0579
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5663)
  AUTHORS   pGreen website
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5663)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5663
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      63..498
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    633..655
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA
                     (truncated)"
     polyA_signal    complement(663..839)
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(958..2607)
                     /label=luciferase
                     /note="firefly luciferase"
     promoter        complement(2618..2963)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     3185..3201
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3208..3226
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    complement(3235..3341)
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(3354..3372)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3393..3409)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3417..3433)
                     /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(3441..3471)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3486..3507)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    3746..3770
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(3861..4449)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4623..5435)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"