Basic Vector Information
- Vector Name:
- pPIGA3
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9375 bp
- Type:
- PiggyBac donor vector
- Replication origin:
- ori
- Source/Author:
- Mitsutake H, Kobayashi J.
- Promoter:
- DmHsp70
pPIGA3 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 ℃.
pPIGA3 vector Sequence
LOCUS 40924_34619 9375 bp DNA circular SYN 18-DEC-2018
DEFINITION PiggyBac donor vector pPIGA3 DNA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9375)
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JOURNAL Unpublished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 9375)
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 9375)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 9375)
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"Unpublished"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..9375
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 96..200
/label=AmpR promoter
CDS 201..1058
/label=AmpR
/note="beta-lactamase"
rep_origin 1232..1820
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 2108..2129
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2144..2174
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2182..2198
/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)."
primer_bind 2206..2222
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
repeat_region 2375..2410
/label=piggyBac 5' inverted terminal repeat
/note="piggyBac 5' inverted terminal repeat"
promoter 3131..3583
/label=hsp70 promoter
/note="Drosophila melanogaster hsp70Bb promoter"
CDS 3682..4278
/label=PuroR
/note="puromycin N-acetyltransferase"
misc_feature 4335..4640
/gene="PGK"
/label=PGK polyadenylation sequence
/note="PGK polyadenylation sequence"
gene 4335..4640
/gene="PGK"
/label=PGK
gene 4658..5313
/gene="A3"
/label=A3
regulatory 4658..4784
/gene="A3"
/label=actin A3
/note="actin A3"
/regulatory_class="promoter"
exon 4785..4853
/gene="A3"
/note="actin A3"
intron 4854..5313
/gene="A3"
/note="actin A3"
CDS 5343..6059
/label=EGFP
/note="enhanced GFP"
polyA_signal 6185..6306
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin complement(6313..6768)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 6795..6899
/gene="bla"
/label=AmpR promoter
promoter 6942..7239
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
misc_feature 8105..8128
/note="piggyBac transposase polyadenylation sequence;
transposase"
repeat_region 8230..8292
/label=piggyBac 3' inverted terminal repeat
/note="piggyBac 3' inverted terminal repeat"
primer_bind complement(8981..8997)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
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