Basic Vector Information
- Vector Name:
- pUASg-HA.attB
- Antibiotic Resistance:
- Ampicillin
- Length:
- 10448 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Bischof J, Bjorklund M, Furger E, Schertel C, Taipale J, Basler K.
- Promoter:
- hsp70
pUASg-HA.attB 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 ℃.
pUASg-HA.attB vector Sequence
LOCUS 40924_44694 10448 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pUASg-HA.attB, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10448)
AUTHORS Bischof J, Bjorklund M, Furger E, Schertel C, Taipale J, Basler K.
TITLE A versatile platform for creating a comprehensive UAS-ORFeome
library in Drosophila
JOURNAL Development 140 (11), 2434-2442 (2013)
PUBMED 23637332
REFERENCE 2 (bases 1 to 10448)
AUTHORS Bischof J, Bjorklund M, Furger E, Schertel C, Taipale J, Basler K.
TITLE Direct Submission
JOURNAL Submitted (13-APR-2013) Institute of Molecular Life Sciences,
University of Zurich, Winterthurerstrasse 190, Zurich 8057,
Switzerland
REFERENCE 3 (bases 1 to 10448)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 10448)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Development"; date: "2013"; volume: "140"; issue: "11"; pages:
"2434-2442"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(13-APR-2013) Institute of Molecular Life Sciences, University of
Zurich, Winterthurerstrasse 190, Zurich 8057, Switzerland"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..10448
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin complement(3..458)
/direction=LEFT
/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 4842..4936
/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 4955..5193
/label=hsp70 promoter
/note="Drosophila melanogaster hsp70Bb promoter"
protein_bind 5230..5354
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter 5379..5409
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 5463..6140
/label=CmR
/note="chloramphenicol acetyltransferase"
CDS 6463..6765
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
protein_bind complement(6809..6933)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
regulatory 6941..6950
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 6980..7069
/label=3xHA
/note="three tandem HA epitope tags"
regulatory 7108..7920
/label=tubulin alpha 1 terminator
/note="tubulin alpha 1 terminator"
/regulatory_class="terminator"
protein_bind 7972..8041
/label=attB
/note="attB site for the phi-C31 integrase (Groth et al.,
2000)"
promoter complement(8260..8278)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(8299..8315)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 8323..8339
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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(8347..8377)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(8392..8413)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(8701..9289)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(9463..10320)
/label=AmpR
/note="beta-lactamase"
promoter complement(10321..10425)
/gene="bla"
/label=AmpR promoter
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