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pFPV25.1-mRFP1 vector (V014138)

Price Information

Cat No. Plasmid Name Availability Add to cart
V014138 pFPV25.1-mRFP1 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:
pFPV25.1-mRFP1
Antibiotic Resistance:
Ampicillin
Length:
5356 bp
Type:
Protein expression
Replication origin:
ori
Host:
E. coli
Promoter:
rpsM
Growth Strain(s):
DH5a
Growth Temperature:
37℃

pFPV25.1-mRFP1 vector Vector Map

pFPV25.1-mRFP15356 bp6001200180024003000360042004800RSF1010 oriTmRFP1Large ribosomal subunit protein bL36AAmpR promoterAmpRori

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.

pFPV25.1-mRFP1 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       V014138                 5356 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014138
VERSION     V014138
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5356)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5356
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            complement(1303..1390)
                     /direction=LEFT
                     /label="RSF1010 oriT"
                     /note="origin of transfer of the broad-host-range plasmid
                     RSF1010 (Scholz et al., 1989)"
     CDS             complement(2023..2697)
                     /label="mRFP1"
                     /note="monomeric derivative of DsRed (Campbell et al.,
                     2002)"
     CDS             complement(3009..3122)
                     /gene="rpmJ"
                     /label="Large ribosomal subunit protein bL36A"
                     /note="Large ribosomal subunit protein bL36A from
                     Escherichia coli (strain K12). Accession#: P0A7Q6"
     promoter        3451..3555
                     /label="AmpR promoter"
     CDS             3556..4413
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      4587..5175
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"