pUAST vector (Cat. No.: V012370)

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Basic Information

Note: pUAST is a P-element-based Drosophila expression vector. It contains five GAL4 binding sites (UAS) for inducible gene expression, a multiple cloning site, P-element ends for genomic integration, the mini-white gene as a selectable marker, and an ampicillin resistance gene for bacterial selection.

Name:: pUAST

Antibiotic Resistance:: Ampicillin

Length:: 8898 bp

Type:: Cloning Vectors

Replication origin:: ori

Source/Author:: Brand AH, Perrimon N.

Copy Number:: High copy number

Promoter:: hsp70

$ 198.2

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

Resources

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

Dragh MA, Al-Allak ZS, Allami ZZ. Cloning and functional analysis of the TERE1 gene using the Gal4-UaS system in S2 cells: A streamlined approach for human gene functional genomics. J Genet Eng Biotechnol. 2025 Sep;23(3):100525. doi: 10.1016/j.jgeb.2025.100525. Epub 2025 Jun 28. PMID: 40854644; PMCID: PMC12269968.

pUAST vector (Cat. No.: V012370) Sequence

LOCUS       pUAST                   8898 bp    DNA     circular SYN 26-DEC-2025
DEFINITION  P element-based vector for Gal4-regulated expression of genes in 
            Drosophila.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8898)
  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 8898)
  AUTHORS   Brand Lab
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8898)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8898)
  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     SGRef: number: 3; type: "Journal Article"
COMMENT     http://www.gurdon.cam.ac.uk/~brandlab/3.html
FEATURES             Location/Qualifiers
     source          1..8898
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          759..763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          1644..1650
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          8488..8502
                     /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            758..4882
                     /label=mini-white
                     /note="This modified version of the white gene lacks part
                     of the first intron."
     gene            759..763
                     /label=mini-white
                     /note="This modified version of the white gene lacks part
                     of the first intron."
     CDS             join(1242..1313,1715..1988,2063..2717,2779..3094,3298..3429,
                     3500..4114)
                     /codon_start=1
                     /gene="white"
                     /product="Drosophila white gene eye color pigment"
                     /label=mini-white
                     /note="This is a modified version of the white gene lacking
                     part of the first intron."
                     /translation="MGQEDQELLIRGGSKHPSAEHLNNGDSGAASQSCINQGFGQAKNY
                     GTLRPPSPPEDSGSGSGQLAENLTYAWHNMDIFGAVNQPGSGWRQLVNRTRGLFCNERH
                     IPAPRKHLLKNVCGVAYPGELLAVMGSSGAGKTTLLNALAFRSPQGIQVSPSGMRLLNG
                     QPVDAKEMQARCAYVQQDDLFIGSLTAREHLIFQAMVRMPRHLTYRQRVARVDQVIQEL
                     SLSKCQHTIIGVPGRVKGLSGGERKRLAFASEALTDPPLLICDEPTSGLDSFTAHSVVQ
                     VLKKLSQKGKTVILTIHQPSSELFELFDKILLMAEGRVAFLGTPSEAVDFFSYVGAQCP
                     TNYNPADFYVQVLAVVPGREIESRDRIAKICDNFAISKVARDMEQLLATKNLEKPLEQP
                     ENGYTYKATWFMQFRAVLWRSWLSVLKEPLLVKVRLIQTTMVAILIGLIFLGQQLTQVG
                     VMNINGAIFLFLTNMTFQNVFATINVFTSELPVFMREARSRLYRCDTYFLGKTIAELPL
                     FLTVPLVFTAIAYPMIGLRAGVLHFFNCLALVTLVANVSTSFGYLISCASSSTSMALSV
                     GPPVIIPFLLFGGFFLNSGSVPVYLKWLSYLSWFRYANEGLLINQWADVEPGEISCTSS
                     NTTCPSSGKVILETLNFSAADLPLDYVGLAILIVSFRVLAYLALRLRARRKE"
     gene            1644..1650
                     /label=mini-white
                     /note="This modified version of the white gene lacks part
                     of the first intron."
     misc_feature    complement(4893..5478)
                     /label=P element 5' end
     misc_feature    5479..5707
                     /label=white linker sequence
                     /note="white linker sequence"
     rep_origin      complement(5713..6301)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(6472..7263,7264..7332))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             complement(6475..7332)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7333..7437)
                     /label=AmpR promoter
     misc_feature    7738..8239
                     /label=white linker sequence
                     /note="white linker sequence"
     misc_feature    complement(8240..8472)
                     /label=P element 3' end
                     /note="P element 3' end"
     protein_bind    8534..8628
                     /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        8647..8885
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"