Basic Vector Information
- Vector Name:
- pPIGA3Fluc
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8127 bp
- Type:
- PiggyBac donor vector
- Replication origin:
- ori
- Source/Author:
- Mitsutake H, Kobayashi J.
- Promoter:
- DmHsp70
pPIGA3Fluc 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 ℃.
pPIGA3Fluc vector Sequence
LOCUS 40924_34624 8127 bp DNA circular SYN 18-DEC-2018
DEFINITION PiggyBac donor vector pPIGA3Fluc DNA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8127)
AUTHORS Mitsutake H, Kobayashi J.
TITLE piggyBac-mediated stable transformation of cultured Bombyx mori
cells using in vitro synthesized transposase mRNA
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 8127)
AUTHORS Mitsutake H, Kobayashi J.
TITLE Direct Submission
JOURNAL Submitted (06-OCT-2010) Contact:Jun Kobayashi Yamaguchi University,
Faculty of Agriculture; Yoshida 1677-1, Yamaguchi, Yamaguchi
753-8515, Japan URL
:http://www.agr.yamaguchi-u.ac.jp/~member/kobayashi/pe.html
REFERENCE 3 (bases 1 to 8127)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 8127)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(06-OCT-2010) Contact:Jun Kobayashi Yamaguchi University, Faculty of
Agriculture"; volume: " Yoshida 1677-1, Yamaguchi, Yamaguchi
753-8515, Japan URL :http"; pages:
"//www.agr.yamaguchi-u.ac.jp/~member/kobayashi/pe.htm"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8127
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 379..395
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
repeat_region 556..591
/label=piggyBac 5' inverted terminal repeat
/note="piggyBac 5' inverted terminal repeat"
promoter 1313..1765
/label=hsp70 promoter
/note="Drosophila melanogaster hsp70Bb promoter"
CDS 1864..2460
/label=PuroR
/note="puromycin N-acetyltransferase"
misc_feature 2517..2822
/gene="PGK"
/label=PGK polyadenylation sequence
/note="PGK polyadenylation sequence"
gene 2517..2822
/gene="PGK"
/label=PGK
gene 2840..3495
/gene="A3"
/label=A3
regulatory 2840..2966
/gene="A3"
/label=actin A3
/note="actin A3"
/regulatory_class="promoter"
exon 2967..3035
/gene="A3"
/note="actin A3"
intron 3036..3495
/gene="A3"
/note="actin A3"
CDS 3525..5174
/label=luciferase
/note="firefly luciferase"
misc_feature 5395..5418
/note="piggyBac transposase polyadenylation sequence;
transposase"
repeat_region 5520..5582
/label=piggyBac 3' inverted terminal repeat
/note="piggyBac 3' inverted terminal repeat"
primer_bind complement(5906..5922)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(5930..5946)
/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(5954..5984)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(5999..6020)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(6308..6896)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7070..7927)
/label=AmpR
/note="beta-lactamase"
promoter complement(7928..8032)
/label=AmpR promoter
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