Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V000916 | pQFlag-USP7 WT 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
- Vector Name:
- pQFlag-USP7 WT puroR
- Antibiotic Resistance:
- Ampicillin
- Length:
- 10495 bp
- Type:
- Mammalian Expression, Retroviral
- Replication origin:
- ori
- Selection Marker:
- Puromycin
- Promoter:
- CMV
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- CMV Forward
- 3' Primer:
- pCDH-Rev
pQFlag-USP7 WT 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.
pQFlag-USP7 WT puroR vector Sequence
LOCUS V000916 10495 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V000916
VERSION V000916
KEYWORDS pQFlag-USP7 WT puroR
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 10495)
AUTHORS Maertens GN, El Messaoudi-Aubert S, Elderkin S, Hiom K, Peters G
TITLE Ubiquitin-specific proteases 7 and 11 modulate Polycomb regulation
of the INK4a tumour suppressor.
JOURNAL EMBO J. 2010 Aug 4;29(15):2553-65. doi: 10.1038/emboj.2010.129. Epub
2010 Jul 2.
PUBMED 20601937
REFERENCE 2 (bases 1 to 10495)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 10495)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1038/emboj.2010.129"; journalName: "EMBO J"; date: "2010-08-4-
4"; volume: "29"; issue: "15"; pages: "2553-65"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..10495
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 5..308
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 309..512
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 513..688
/label="5' LTR (truncated)"
/note="truncated long terminal repeat from Moloney murine
sarcoma virus"
misc_feature 751..950
/label="MMLV Psi"
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
CDS 1151..1567
/label="gag (truncated)"
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
enhancer 1604..1907
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 1908..2111
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 2108..2132
/label="LNCX"
/note="Human CMV promoter, forward primer"
CDS 2268..2291
/label="FLAG"
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS 2298..5603
/gene="USP7"
/label="Ubiquitin carboxyl-terminal hydrolase 7"
/note="Ubiquitin carboxyl-terminal hydrolase 7 from Homo
sapiens. Accession#: Q93009"
misc_feature 5622..6208
/label="IRES2"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 6228..6824
/label="PuroR"
/note="puromycin N-acetyltransferase"
LTR 7201..7626
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
Moloney murine leukemia virus"
primer_bind complement(7729..7751)
/label="pGEX 3'"
/note="pGEX vectors, reverse primer"
promoter 7892..8221
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
primer_bind 8243..8262
/label="pRS-marker"
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(8343..8360)
/label="L4440"
/note="L4440 vector, forward primer"
rep_origin complement(8514..9102)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(9276..10133)
/label="AmpR"
/note="beta-lactamase"
promoter complement(10134..10238)
/label="AmpR promoter"
primer_bind 10306..10324
/label="pBRforEco"
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"