pJG4-5 vector (V010260)

COA reports Sequencing Results MSDS Download GenBank File(.gb)

Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V010260	pJG4-5	In stock, 1 week for quality controls	Buy one, get one free!

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

Basic Vector Information

Vector Name:: pJG4-5

Antibiotic Resistance:: Ampicillin

Length:: 6449 bp

Type:: Yeast Plasmids

Replication origin:: ori

Source/Author:: Gyuris J, Golemis E, Chertkov H, Brent R.

Copy Number:: High copy number

Promoter:: GAL1

pJG4-5 vector Map

Copy Sequence View Map

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.

pJG4-5 vector Sequence

LOCUS       pJG4-5.        6449 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Yeast two-hybrid vector with a GAL1 promoter, for fusing the B42 
            transcriptional activator to a partner protein. Also known as 
            pB42AD.
ACCESSION   .
VERSION     .
KEYWORDS    pJG4-5
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6449)
  AUTHORS   Gyuris J, Golemis E, Chertkov H, Brent R.
  TITLE     Cdi1, a human G1 and S phase protein phosphatase that associates 
            with Cdk2.
  JOURNAL   Cell 1993;75:791-803.
  PUBMED    8242750
REFERENCE   2  (bases 1 to 6449)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6449)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell"; 
            date: "1993"; volume: "75"; pages: "791-803"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6449
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        81..522
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     CDS             534..536
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             546..566
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     CDS             573..809
                     /label=B42 transcriptional activator
                     /note="yeast transcriptional activator cretaed from E. coli
                     genomic DNA fragments (Ma and Ptashne, 1987)"
     CDS             816..842
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
     misc_feature    849..866
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     terminator      1004..1191
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     rep_origin      1713..3055
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(3419..4090)
                     /label=TRP1
                     /note="phosphoribosylanthranilate isomerase, required for 
                     tryptophan biosynthesis"
     promoter        complement(4091..4192)
                     /label=TRP1 promoter
     promoter        4295..4399
                     /label=AmpR promoter
     CDS             4400..5257
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      5431..6019
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6307..6328
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6343..6373
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6381..6397
                     /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     6405..6421
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"