pBV220-mCherry-Linker-NDH2 vector (V013643)

Price Information

Cat No. Plasmid Name Availability Add to cart
V013643 pBV220-mCherry-Linker-NDH2 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:
pBV220-mCherry-Linker-NDH2
Antibiotic Resistance:
Ampicillin
Length:
5661 bp
Type:
Protein expression
Replication origin:
ori
Host:
E. coli
Promoter:
PL,PR
Growth Strain(s):
DH5a
Growth Temperature:
30℃

pBV220-mCherry-Linker-NDH2 vector Map

pBV220-mCherry-Linker-NDH25661 bp60012001800240030003600420048005400mCherryType II NADH:quinone oxidoreductaserrnB T1 terminatorrrnB T2 terminatorAmpR promoterAmpRorilambda repressor

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.

pBV220-mCherry-Linker-NDH2 vector Sequence

LOCUS       V013643                 5661 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V013643
VERSION     V013643
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5661)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5661
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             434..1141
                     /label="mCherry"
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     CDS             1145..2401
                     /gene="ndh"
                     /label="Type II NADH:quinone oxidoreductase"
                     /note="Type II NADH:quinone oxidoreductase from Escherichia
                     coli (strain K12). Accession#: P00393"
     terminator      2660..2746
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2838..2865
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        2883..2974
                     /label="AmpR promoter"
     CDS             2975..3832
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      4006..4594
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(4951..5661)
                     /label="lambda repressor"
                     /note="phage lambda repressor"