pGTMV-T159K vector (V014196)

Price Information

Cat No. Plasmid Name Availability Add to cart
V014196 pGTMV-T159K In stock, 1 week for quality controls

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

Basic Vector Information

Vector Name:
pGTMV-T159K
Antibiotic Resistance:
Kanamycin
Length:
9669 bp
Type:
Protein expression
Replication origin:
pSa ori
Host:
Plants
Copy Number:
Low
Promoter:
35s
Growth Strain(s):
DH5a
Growth Temperature:
30℃

pGTMV-T159K vector Map

pGTMV-T159K9669 bp4008001200160020002400280032003600400044004800520056006000640068007200760080008400880092009600TMV OmegaMovement proteinCapsid proteinRB T-DNA repeatoriKanRpSa oriLB T-DNA repeat

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.

pGTMV-T159K vector Sequence

LOCUS       V014196                 9669 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014196
VERSION     V014196
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9669)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9669
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    101..156
                     /label="TMV Omega"
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     CDS             4992..5795
                     /gene="MP"
                     /label="Movement protein"
                     /note="Movement protein from Tobacco mosaic virus (strain
                     vulgare). Accession#: P03583"
     CDS             5801..6274
                     /gene="CP"
                     /label="Capsid protein"
                     /note="Capsid protein from Tobacco mosaic virus (strain
                     vulgare). Accession#: P69687"
     misc_feature    6774..6798
                     /label="RB T-DNA repeat"
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(6917..7505)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7679..8491)
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     rep_origin      8782..9217
                     /label="pSa ori"
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    9368..9392
                     /label="LB T-DNA repeat"
                     /note="left border repeat from nopaline C58 T-DNA"