pGreenII 0800-LUC vector (V010545)

Price Information

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

Basic Vector Information

Helper plasmid eg: pSoup could faciliate its replication in Agrobacterium

Vector Name:
pGreenII 0800-LUC
Antibiotic Resistance:
Kanamycin
Length:
6382 bp
Type:
Plant Expression Vectors, Promoter Reporter Vector
Replication origin:
pSa ori
Host:
Plants
Selection Marker:
Luc
Promoter:
CaMV 35S
Growth Strain(s):
Stbl3
Growth Temperature:
37℃

pGreenII 0800-LUC vector Vector Map

pGreenII 0800-LUC6382 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300LB T-DNA repeatCaMV 35S promoterRlucCaMV poly(A) signalM13 fwdT7 promoterMCSluciferaseCaMV poly(A) signalRB T-DNA repeatoriKanRpSa ori

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 0800-LUC vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_22636        6382 bp DNA     circular SYN 13-JAN-2022
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6382)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6382)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6382
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    542..564
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA
                     (truncated)"
     promoter        626..971
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             982..1914
                     /label=Rluc
                     /note="luciferase from the anthozoan coelenterate Renilla 
                     reniformis (sea pansy)"
     polyA_signal    1974..2150
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     2314..2330
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2337..2355
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    complement(2364..2469)
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     CDS             2479..4128
                     /label=luciferase
                     /note="firefly luciferase"
     polyA_signal    4188..4364
                     /label=CaMV poly(A) signal
                     /note="CaMV poly(A) signal"
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    4374..4398
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4489..5077)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5251..6063)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      6354..6382
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"