Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V017511 | PiggyBac PlayBack | In stock, instant shipping |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
To all for PiggyBac gene insertion into Genome
- Vector Name:
- PiggyBac PlayBack
- Antibiotic Resistance:
- Ampicillin
- Length:
- 3237 bp
- Copy Number:
- High copy number
- Growth Strain(s):
- DH10B
- Growth Temperature:
- 37℃
PiggyBac PlayBack 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.
PiggyBac PlayBack vector Sequence
LOCUS Exported 3237 bp DNA circular SYN 03-DEC-2024
DEFINITION To all for PiggyBac gene insertion into Genome.
ACCESSION .
VERSION .
KEYWORDS PiggyBac PlayBack
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3237)
AUTHORS Foran G, Hallam RD, Megaly M, Turgambayeva A, Necakov A
TITLE PlayBack cloning: simple, reversible, cost-effective cloning for the
combinatorial assembly of complex expression constructs.
JOURNAL Biotechniques. 2023 Oct;75(4):168-178. doi: 10.2144/btn-2023-0042.
Epub 2023 Oct 10.
PUBMED 37815818
REFERENCE 2 (bases 1 to 3237)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.2144/btn-2023-0042"; journalName: "Biotechniques"; date:
"2023-10"; volume: "75"; issue: "4"; pages: "168-178"
FEATURES Location/Qualifiers
source 1..3237
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(29..51)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 151..170
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
repeat_region 194..228
/label=piggyBac left (5') inverted repeat
/note="piggyBac transposon-specific inverted terminal
repeat sequence (ITR)"
repeat_region 931..993
/label=piggyBac right (3') inverted repeat
/note="piggyBac transposon-specific inverted terminal
repeat sequence (ITR)"
primer_bind complement(1016..1032)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1016..1032)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(1029..1051)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 1040..1056
/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(1064..1094)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1109..1130
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(1418..2006)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1498..1517)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2177..3037)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 2800..2819
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3038..3142)
/gene="bla"
/label=AmpR promoter
primer_bind 3210..3228
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"