Basic Vector Information
pUASTattB vector Vector Map
Plasmid Resuspension 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 ℃.
pUASTattB vector Sequence
LOCUS 40924_44774 8489 bp DNA circular SYN 18-DEC-2018 DEFINITION Cloning and transformation vector pUASTattB, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8489) AUTHORS Bischof J, Maeda RK, Hediger M, Karch F, Basler K. TITLE An optimized transgenesis system for Drosophila using germ-line-specific phiC31 integrases JOURNAL Proc. Natl. Acad. Sci. U.S.A. 104 (9), 3312-3317 (2007) PUBMED 17360644 REFERENCE 2 (bases 1 to 8489) AUTHORS Bischof J, Maeda RK, Hediger M, Karch F, Basler K. TITLE Direct Submission JOURNAL Submitted (12-JAN-2007) Institute of Molecular Biology, University of Zurich, Winterthurerstrasse 190, Zurich 8057, Switzerland REFERENCE 3 (bases 1 to 8489) TITLE Direct Submission REFERENCE 4 (bases 1 to 8489) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Proc. Natl. Acad. Sci. U.S.A."; date: "2007"; volume: "104"; issue: "9"; pages: "3312-3317" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (12-JAN-2007) Institute of Molecular Biology, University of Zurich, Winterthurerstrasse 190, Zurich 8057, Switzerland" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..8489 /mol_type="other DNA" /organism="synthetic DNA construct" rep_origin 4..459 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind 600..616 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 623..641 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" gene 666..4802 /label=mini-white /note="This modified version of the white gene lacks part of the first intron." protein_bind 4809..4842 /label=loxP /note="Cre-mediated recombination occurs in the 8-bp core sequence (ATGTATGC) (Shaw et al., 2021)." protein_bind 4875..4969 /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 4988..5226 /label=hsp70 promoter /note="Drosophila melanogaster hsp70Bb promoter" misc_feature 5231..5285 /label=multiple cloning site /note="multiple cloning site" intron 5363..5428 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" CDS 5558..5578 /codon_start=1 /label=SV40 NLS /note="nuclear localization signal of SV40 (simian virus 40) large T antigen" /translation="PKKKRKV" polyA_signal 5850..5984 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" protein_bind 6013..6082 /label=attB /note="attB site for the phi-C31 integrase (Groth et al., 2000)" promoter complement(6301..6319) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(6340..6356) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(6364..6380) /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(6388..6418) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(6433..6454) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(6742..7330) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(7504..8361) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(8362..8466) /label=AmpR promoter
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