Basic Vector Information
pVALIUM10-roe 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 ℃.
pVALIUM10-roe vector Sequence
LOCUS 40924_45883 9240 bp DNA circular SYN 18-DEC-2018
DEFINITION Overexpression vector pVALIUM10-roe, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9240)
AUTHORS Ni J-Q., Liu L-P., Perkins LA, Perrimon N.
TITLE Vectors for transgenic flies
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 9240)
AUTHORS Ni J-Q., Liu L-P., Perkins LA, Perrimon N.
TITLE Direct Submission
JOURNAL Submitted (22-FEB-2010) Genetics, Harvard Medical School, 77 Avenue
Louis Pasteur, Boston, MA 02115, USA
REFERENCE 3 (bases 1 to 9240)
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 9240)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 9240)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(22-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 GU931386.1.
FEATURES Location/Qualifiers
source 1..9240
/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=vermillion
/note="vermillion"
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="ADE08348.1"
/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..3377
/label=gypsy insulator
/note="chromatin insulator from Drosophila"
protein_bind 3395..3428
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
protein_bind 3441..3535
/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 3551..3584
/label=LoxP
/note="LoxP"
protein_bind complement(3551..3584)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
misc_feature 3591..3700
/label=5XUAS
/note="5XUAS"
protein_bind 3597..3691
/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 3715..3953
/label=hsp70 promoter
/note="Drosophila melanogaster hsp70Bb promoter"
protein_bind 3998..4122
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter 4147..4177
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 4231..4887
/label=CmR
/note="chloramphenicol acetyltransferase"
CDS 5232..5534
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
protein_bind complement(5578..5702)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
intron 5763..5909
/note="ftz intron"
intron 6009..6074
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 6204..6224
/label=SV40 NLS
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
polyA_signal 6496..6630
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
protein_bind 6643..7028
/label=gypsy insulator
/note="chromatin insulator from Drosophila"
promoter complement(7088..7106)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(7127..7143)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(7151..7167)
/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(7175..7205)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(7220..7241)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(7529..8117)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(8291..9148)
/label=AmpR
/note="beta-lactamase"
promoter complement(9149..9240)
/label=AmpR promoter
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