Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V012836 | Opie2-PuroR | In stock, 1 week for quality controls |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
Expresses PuroR and dsRed.
- Vector Name:
- Opie2-PuroR
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8646 bp
- Type:
- Insect Cell Vectors
- Replication origin:
- ori
- Source/Author:
- Omar Akbari
- Promoter:
- IE1
- Fusion Tag:
- DsRed1
- Growth Strain(s):
- DH5alpha
- Growth Temperature:
- 37℃
Opie2-PuroR vector Map
Plasmid 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 ℃.
6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it
General Plasmid Transform Protocol
1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.
2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.
3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.
4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.
5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.
Opie2-PuroR vector Sequence
LOCUS Opie2-PuroR. 8646 bp DNA circular SYN 15-JAN-2024
DEFINITION Expresses PuroR and dsRed..
ACCESSION .
VERSION .
KEYWORDS Opie2-PuroR
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8646)
AUTHORS Kandul NP, Liu J, Hsu AD, Hay BA, Akbari OS
TITLE A drug-inducible sex-separation technique for insects.
JOURNAL Nat Commun. 2020 Apr 30;11(1):2106. doi: 10.1038/s41467-020-16020-2.
PUBMED 32355156
REFERENCE 2 (bases 1 to 8646)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Commun."; date: "2020-04-30"; pages: "
10.1038/s41467-020-16020-2"
FEATURES Location/Qualifiers
source 1..8646
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 60..77
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 194..215
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 230..260
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 268..284
/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 292..308
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
repeat_region 489..523
/label=piggyBac left (5') inverted repeat
/note="piggyBac transposon-specific inverted terminal
repeat sequence (ITR)"
polyA_signal complement(1212..1333)
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
CDS complement(1457..2134)
/label=DsRed1
/note="wild-type DsRed"
promoter complement(2145..2736)
/label=IE1 promoter
/note="promoter of the ie1 gene from the baculovirus
Autographa californica"
enhancer complement(2740..3222)
/label=hr5 enhancer
/note="baculovirus early transcription enhancer"
CDS 3785..4381
/label=PuroR
/note="puromycin N-acetyltransferase"
polyA_signal 4402..4536
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
repeat_region 5572..5634
/label=piggyBac right (3') inverted repeat
/note="piggyBac transposon-specific inverted terminal
repeat sequence (ITR)"
primer_bind complement(6338..6354)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6563..6582)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6682..6704
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(6742..6760)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 6828..6932
/label=AmpR promoter
CDS 6933..7790
/label=AmpR
/note="beta-lactamase"
rep_origin 7964..8552
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"