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pBE2R(cm) vector (V015254)

Price Information

Cat No. Plasmid Name Availability Add to cart
V015254 pBE2R(cm) 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:
pBE2R(cm)
Antibiotic Resistance:
Ampicillin
Length:
7700 bp
Type:
Protein expression, Secretory expression
Replication origin:
ori
Host:
Bacillus subtilis
Selection Marker:
Chl
Promoter:
p43
Growth Strain(s):
Stbl3
Growth Temperature:
30℃

pBE2R(cm) vector Map

Copy Sequence View Map
pBE2R(cm)7700 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600690072007500AmpRoriChloramphenicol acetyltransferaserepB6xHisSP6 promoterAmpR promoter

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.

pBE2R(cm) vector Sequence

LOCUS       V015254                 7700 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V015254
VERSION     V015254
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7700)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7700
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      891..1479
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(3531..4178)
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     CDS             complement(4504..5505)
                     /label="repB"
                     /note="RepB replication protein"
     CDS             6624..6641
                     /label="6xHis"
                     /note="6xHis affinity tag"
     promoter        complement(6652..6670)
                     /label="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     promoter        7455..7559
                     /label="AmpR promoter"
     CDS             join(7560..7700,1..717)
                     /label="AmpR"
                     /note="beta-lactamase"