Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V010380 | pRetroX-TetOne-Puro | 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
- Vector Name:
- pRetroX-TetOne-Puro
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9463 bp
- Type:
- Viral Expression & Packaging Vectors
- Replication origin:
- ori
- Source/Author:
- Clontech
- Copy Number:
- High copy number
- Promoter:
- hPGK
pRetroX-TetOne-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.
pRetroX-TetOne-Puro vector Sequence
LOCUS pRetroX-TetOne-P 9463 bp DNA circular SYN 01-JAN-1980
DEFINITION All-in-one retroviral vector with a puromycin resistance marker, for
strong inducible expression of genes using the Tet-On(R) system.
ACCESSION .
VERSION .
KEYWORDS pRetroX-TetOne-Puro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9463)
AUTHORS Clontech
TITLE Direct Submission
REFERENCE 2 (bases 1 to 9463)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..9463
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 164..543
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
misc_feature 974..1331
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
CDS 1405..1812
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
misc_feature 1822..2193
/label=pol region
/note="Moloney murine leukemia virus (MMLV) pol region
containing the splice acceptor site"
polyA_signal 2298..2432
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
misc_feature 2607..2638
/label=MCS
/note="MCS"
/note="multiple cloning site"
promoter complement(2639..3006)
/label=TRE3GS promoter
/note="3rd-generation Tet-responsive promoter that can be
activated by binding of Tet-On(R) 3G, modified to eliminate
binding sites for endogenous mammalian transcription
factors"
promoter 3023..3533
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
CDS 3552..4295
/label=Tet-On(R) 3G
/note="modified rtTA protein that binds tightly to
promoters containing the tet operator in the presence of
doxycycline"
promoter 4316..4645
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
CDS 4654..5250
/label=PuroR
/note="puromycin N-acetyltransferase"
misc_feature 5271..5674
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 5835..6047
/label=3'-LTR del in U3
/note="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
Moloney murine leukemia virus"
polyA_signal 6690..6811
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(6834..6850)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 7325..7429
/label=AmpR promoter
CDS 7430..8287
/label=AmpR
/note="beta-lactamase"
rep_origin 8461..9049
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 9337..9358
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 9373..9403
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 9411..9427
/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 9435..9451
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"