pBV220-OmpA-JI996(Salmonella)-PD1 vector (V014351)

Price Information

Cat No. Plasmid Name Availability Add to cart
V014351 pBV220-OmpA-JI996(Salmonella)-PD1 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-OmpA-JI996(Salmonella)-PD1
Antibiotic Resistance:
Ampicillin
Length:
5224 bp
Type:
Protein expression
Replication origin:
ori
Host:
E. coli
Promoter:
PL,PR
Growth Strain(s):
DH5a
Growth Temperature:
30℃

pBV220-OmpA-JI996(Salmonella)-PD1 vector Vector Map

pBV220-OmpA-JI996(Salmonella)-PD15224 bp6001200180024003000360042004800M13 revOmpA signal peptideProgrammed cell death protein 1rrnB T1 terminatorrrnB T2 terminatorAmpR promoterAmpRorilambda repressor (ts)

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-OmpA-JI996(Salmonella)-PD1 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       V014351                 5224 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014351
VERSION     V014351
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5224)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5224
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     396..412
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     sig_peptide     458..520
                     /label="OmpA signal peptide"
                     /note="signal peptide from the E. coli outer membrane
                     protein OmpA"
     CDS             1082..1945
                     /gene="Pdcd1"
                     /label="Programmed cell death protein 1"
                     /note="Programmed cell death protein 1 from Mus musculus.
                     Accession#: Q02242"
     terminator      2177..2263
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2355..2382
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        2401..2492
                     /label="AmpR promoter"
     CDS             2493..3350
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      3524..4112
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(4467..5177)
                     /label="lambda repressor (ts)"
                     /note="temperature-sensitive variant of the phage lambda
                     repressor"