Basic Vector Information
pVALIUM1 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 ℃.
pVALIUM1 vector Sequence
LOCUS 40924_45873 6742 bp DNA circular SYN 18-DEC-2018 DEFINITION RNAi cloning vector pVALIUM1, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6742) AUTHORS Ni J-Q., Liu L-P., Perkins LA, Perrimon N. TITLE Vectors for transgenic flies JOURNAL Unpublished REFERENCE 2 (bases 1 to 6742) AUTHORS Ni J-Q., Liu L-P., Perkins LA, Perrimon N. TITLE Direct Submission JOURNAL Submitted (02-FEB-2010) Genetics, Harvard Medical School, 77 Avenue Louis Pasteur, Boston, MA 02115, USA REFERENCE 3 (bases 1 to 6742) AUTHORS Hu Y, Perkins LA. TITLE Direct Submission JOURNAL Submitted (20-JUN-2012) Genetics, Harvard Medical School, 77 Avenue Louis Pasteur, Boston, MA 02115, USA REFERENCE 4 (bases 1 to 6742) TITLE Direct Submission REFERENCE 5 (bases 1 to 6742) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Unpublished" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (02-FEB-2010) Genetics, Harvard Medical School, 77 Avenue Louis Pasteur, Boston, MA 02115, USA" COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted (20-JUN-2012) Genetics, Harvard Medical School, 77 Avenue Louis Pasteur, Boston, MA 02115, USA" COMMENT SGRef: number: 4; type: "Journal Article" COMMENT On Jun 20, 2012 this sequence version replaced GU931383.1. FEATURES Location/Qualifiers source 1..6742 /mol_type="other DNA" /organism="synthetic DNA construct" rep_origin complement(6..461) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind 603..619 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 626..644 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" misc_feature 664..2543 /label=vermilion /note="vermilion" CDS complement(join(834..951,1028..1121,1213..1819,1874..2007, 2068..2228,2302..2327)) /codon_start=1 /product="vermillion" /label=vermillion /note="selectable marker" /protein_id="ADE08345.1" /translation="MSCPYAGNGNDHDDSAVPLTTEVGKIYGEYLMLDKLLDAQCMLSE EDKRPVHDEHLFIITHQAYELWFKQIIFEFDSIRDMLDAEVIDETKTLEIVKRLNRVVL ILKLLVDQVPILETMTPLDFMDFRKYLAPASGFQSLQFRLIENKLGVLTEQRVRYNQKY SDVFSDEEARNSIRNSEKDPSLLELVQRWLERTPGLEESGFNFWAKFQESVDRFLEAQV QSAMEEPVEKAKNYRLMDIEKRREVYRSIFDPAVHDALVRRGDRRFSHRALQGAIMITF YRDEPRFSQPHQLLTLLMDIDSLITKWRYNHVIMVQRMIGSQQLGTGGSSGYQYLRSTL SDRYKVFLDLFNLSTFLIPREAIPPLDETIRKKLINKSV" protein_bind 2640..2709 /label=attB /note="attB site for the phi-C31 integrase (Groth et al., 2000)" protein_bind 2947..2980 /label=loxP /note="Cre-mediated recombination occurs in the 8-bp core sequence (ATGTATGC) (Shaw et al., 2021)." protein_bind 2993..3087 /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)" misc_recomb 3103..3136 /label=LoxP /note="LoxP" protein_bind complement(3103..3136) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 3143..3252 /label=5XUAS /note="5XUAS" protein_bind 3149..3243 /label=5X UAS /bound_moiety="GAL4" /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 3267..3505 /label=hsp70 promoter /note="Drosophila melanogaster hsp70Bb promoter" misc_feature 3519..3524 /label=multiple cloning site /note="multiple cloning site" intron 3547..3620 /note="white intron" misc_feature 3620..3625 /label=multiple cloning site /note="multiple cloning site" intron 3669..3815 /note="ftz intron" intron 3915..3980 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" CDS 4110..4130 /label=SV40 NLS /note="nuclear localization signal of SV40 (simian virus 40) large T antigen" polyA_signal 4402..4536 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" promoter complement(4557..4575) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(4596..4612) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(4620..4636) /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(4644..4674) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(4689..4710) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(4998..5586) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(5760..6617) /label=AmpR /note="beta-lactamase" promoter complement(6618..6722) /label=AmpR promoter
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