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Price Information

Cat No. Plasmid Name Availability Buy one, get one free! (?)
V012457 pJET1.2 In stock, instant shipping

Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

pJET1.2, as a blunt-end cloning vector, is used to ligate PCR products of target genes and subsequently transformed into Escherichia coli DH5α. This provides a qualified gene clone template for subsequent subcloning into expression vectors.

Vector Name:
pJET1.2
Antibiotic Resistance:
Ampicillin
Length:
3005 bp
Type:
Cloning Vectors
Replication origin:
ori
Source/Author:
Thermo Scientific (Fermentas)
Copy Number:
High copy number
Promoter:
lac UV5
Growth Strain(s):
DH10B

pJET1.2 vector Map

Copy Sequence View Map
pJET1.23005 bp6001200180024003000T7 promoterMCSlac operatorlac UV5 promoterCAP binding siteoriAmpRAmpR 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.

References

  • Nawawi O, Abdullah MP, Yusuf CYL. A streamlined strategy for self-production of a commercial positive selection vector, the pJET1.2/blunt cloning vector, using common laboratory E. coli strains. 3 Biotech. 2023 Jul;13(7):224. doi: 10.1007/s13205-023-03647-3. Epub 2023 Jun 6. PMID: 37292140; PMCID: PMC10244300.

pJET1.2 vector Sequence

LOCUS       Exported                3005 bp DNA     circular SYN 03-SEP-2024
DEFINITION  Positive selection cloning vector with a lethal insert that allows 
            for efficient recovery of blunt-ended PCR products.
ACCESSION   .
VERSION     .
KEYWORDS    pJET1.2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3005)
  AUTHORS   Thermo Scientific (Fermentas)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 3005)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 3005)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     Linearize at Eco32I (EcoRV) to insert a blunt PCR product.
FEATURES             Location/Qualifiers
     source          1..3005
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          372..402
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        305..323
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    328..453
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site with a central Eco32I (EcoRV)
                     site"
     protein_bind    complement(813..829)
                     /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)."
     promoter        complement(837..867)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     protein_bind    complement(882..903)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1194..1782)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1956..2813)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(2814..2918)
                     /label=AmpR promoter