pJL-TRBO-G vector (V006590)

Price Information

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V006590 pJL-TRBO-G In stock, instant shipping

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

Basic Vector Information

Recommended culture condition: DH5α competent cells,kanamycin 50ug/ml,culture temperature 30℃(not 37℃)

Vector Name:
pJL-TRBO-G
Antibiotic Resistance:
Kanamycin
Length:
11329 bp
Type:
Plant Expression
Replication origin:
oriV
Selection Marker:
Gentamicin ; Rifampicin
Copy Number:
Low Copy

pJL-TRBO-G vector Map

pJL-TRBO-G11329 bp500100015002000250030003500400045005000550060006500700075008000850090009500100001050011000minimal CaMV 35S promoterTMV OmegaMovement proteinCycle 3 GFPRB T-DNA repeattrfAKanRoriVLB T-DNA repeatKS primer

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.

pJL-TRBO-G vector Sequence

LOCUS       V006590                11329 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V006590
VERSION     V006590
KEYWORDS    pJL-TRBO-G
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 11329)
  AUTHORS   Lindbo JA
  TITLE     TRBO: a high-efficiency tobacco mosaic virus RNA-based
            overexpression vector.
  JOURNAL   Plant Physiol. 2007 Dec;145(4):1232-40. Epub 2007 Aug 24.
   PUBMED   17720752
REFERENCE   2  (bases 1 to 11329)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 11329)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant
            Physiol."; date: "2007-12"; volume: "145(4)"; pages: "1232-40. Epub
            2007 Aug 24"
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..11329
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..46
                     /label="minimal CaMV 35S promoter"
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     misc_feature    58..113
                     /label="TMV Omega"
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     CDS             4949..5752
                     /gene="MP"
                     /label="Movement protein"
                     /note="Movement protein from Tobacco mosaic virus (strain
                     vulgare). Accession#: P03583"
     CDS             5811..6527
                     /label="Cycle 3 GFP"
                     /note="Cycle 3 GFP (Crameri et al., 1996)"
     misc_feature    7137..7161
                     /label="RB T-DNA repeat"
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(7319..8464)
                     /label="trfA"
                     /note="trans-acting replication protein that binds to and
                     activates oriV"
     CDS             complement(8766..9557)
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(9766..10401)
                     /direction=LEFT
                     /label="oriV"
                     /note="incP origin of replication"
     misc_feature    10481..10505
                     /label="LB T-DNA repeat"
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     10586..10602
                     /label="KS primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"