Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V014710 | PiggyBac-EF1a-MCS-EF1a-Puro | In stock, instant shipping |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
Core Function: A mammalian expression plasmid designed for stable genomic integration of target genes using the PiggyBac transposon system. Key Components: 1. Promoter: Uses the EF-1α (Elongation Factor 1-alpha) promoter for constitutive, medium-to-high expression in most mammalian cell types 2. Selection Marker: Puromycin resistance gene (PuroR) under the SV40 poly(A) signal, enabling antibiotic selection of stably integrated cells 3. Backbone: Includes an Ampicillin resistance gene (AmpR) and pUC origin for bacterial propagation
- Vector Name:
- PiggyBac-EF1a-MCS-EF1a-Puro
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6449 bp
- Type:
- Protein expression, Transposition
- Replication origin:
- ori
- Host:
- Mammalian cells
- Selection Marker:
- Puro
- Promoter:
- EF-1α
- Growth Temperature:
- 37℃
PiggyBac-EF1a-MCS-EF1a-Puro 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-EF1a-MCS-EF1a-Puro vector Sequence
LOCUS Exported 6449 bp DNA circular SYN 24-OCT-2025
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6449)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 6449)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6449
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..105
/label=AmpR promoter
rep_origin complement(131..586)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 728..744
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
LTR 870..1104
/label=3' ITR
/note="piggyBac 3' inverted terminal repeat"
promoter 1442..1645
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 1786..1997
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
LTR 2010..2278
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from human
T-cell leukemia virus (HTLV) type 1"
CDS 2313..2909
/codon_start=1
/label=PuroR
/note="puromycin N-acetyltransferase"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
polyA_signal 3020..3152
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
LTR 3629..3941
/label=5' ITR
/note="piggyBac 5' inverted terminal repeat"
primer_bind complement(4428..4444)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 4452..4468
/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 4476..4506
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4521..4542
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4830..5418)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5592..6449)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"