pKSV7 vector (V014085)

Price Information

Cat No. Plasmid Name Availability Add to cart
V014085 pKSV7 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:
pKSV7
Antibiotic Resistance:
Ampicillin
Length:
7139 bp
Replication origin:
ori
Host:
E. coli
Growth Strain(s):
DH5a
Growth Temperature:
37℃

pKSV7 vector Vector Map

pKSV77139 bp30060090012001500180021002400270030003300360039004200450048005100540057006000630066006900CAP binding sitelac promoterlac operatorM13 revMCSM13 fwdAmpR promoterAmpRoriChloramphenicol acetyltransferaserRNA adenine N-6-methyltransferase

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.

pKSV7 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       V014085                 7139 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014085
VERSION     V014085
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7139)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7139
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    55..76
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        91..121
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    129..145
                     /label="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     153..169
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    complement(182..238)
                     /label="MCS"
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(239..255)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        729..833
                     /label="AmpR promoter"
     CDS             834..1691
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      1865..2453
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(2989..3636)
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     CDS             4167..4898
                     /gene="ermC"
                     /label="rRNA adenine N-6-methyltransferase"
                     /note="rRNA adenine N-6-methyltransferase from
                     Staphylococcus aureus. Accession#: P02979"