pUAST vector (V012370)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V012370	pUAST	In stock, instant shipping	Buy one, get one free!

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

Basic Vector Information

Vector Name:: pUAST

Antibiotic Resistance:: Ampicillin

Length:: 8897 bp

Type:: Cloning Vectors

Replication origin:: ori

Source/Author:: Brand AH, Perrimon N.

Copy Number:: High copy number

Promoter:: hsp70

pUAST 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.

pUAST vector Sequence

LOCUS       pUAST.        8897 bp DNA     circular SYN 01-JAN-1980
DEFINITION  P element-based vector for Gal4-regulated expression of genes in 
            Drosophila.
ACCESSION   .
VERSION     .
KEYWORDS    pUAST
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8897)
  AUTHORS   Brand AH, Perrimon N.
  TITLE     Targeted gene expression as a means of altering cell fates and 
            generating dominant phenotypes.
  JOURNAL   Development 1993;118:401-15.
  PUBMED    8223268
REFERENCE   2  (bases 1 to 8897)
  AUTHORS   Brand Lab
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8897)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Development"; date: "1993"; volume: "118"; pages: "401-15"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     http://www.gurdon.cam.ac.uk/~brandlab/3.html
FEATURES             Location/Qualifiers
     source          1..8897
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..46
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     intron          124..189
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             319..339
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     polyA_signal    611..745
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     gene            763..4899
                     /label=mini-white
                     /note="This modified version of the white gene lacks part
                     of the first intron."
     misc_feature    complement(4910..5495)
                     /label=P element 5' end
     misc_feature    5496..5724
                     /label=white linker sequence
                     /note="white linker sequence"
     rep_origin      complement(5730..6318)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6492..7349)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7350..7454)
                     /label=AmpR promoter
     misc_feature    7755..8255
                     /label=white linker sequence
                     /note="white linker sequence"
     misc_feature    complement(8256..8488)
                     /label=P element 3' end
                     /note="P element 3' end"
     protein_bind    8533..8627
                     /label=5X UAS
                     /note="five tandem copies of the 'ScaI site' 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     promoter        8646..8884
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"