Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V011112 | pHom-Mem1 | 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
Proteins fused with FKBP12F36V (DmrB), a mutant of the 12-kDa FK506-binding protein (FKBP12), can be rapidly degraded in cultured cells and mice in the presence of cell-permeable heterobifunctional degraders called dTAGs.
- Vector Name:
- pHom-Mem1
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5726 bp
- Type:
- Mammalian Expression Vectors
- Replication origin:
- ori
- Source/Author:
- Clontech
- Copy Number:
- High copy number
- Promoter:
- SV40
- Growth Strain(s):
- DH10B
- Growth Temperature:
- 37℃
pHom-Mem1 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.
References
- Yesbolatova A, Kanemaki MT. TAGing for destruction. Nat Chem Biol. 2018 May;14(5):414-415. doi: 10.1038/s41589-018-0024-5. PMID: 29581583.
pHom-Mem1 vector Sequence
LOCUS pHom-Mem1. 5726 bp DNA circular SYN 01-JAN-1980
DEFINITION Mammalian vector encoding FKBP homodimerizer domains, for creating
membrane-associated fusion proteins that can be dimerized with a
drug. Also known as pC4M-FV2E.
ACCESSION .
VERSION .
KEYWORDS pHom-Mem1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5726)
AUTHORS Clontech
TITLE Direct Submission
REFERENCE 2 (bases 1 to 5726)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT Clone into the XbaI site to fuse the DmrB domains to the C-terminus
of the partner protein, or into the SpeI site to fuse the DmrB
domains to the N-terminus of the partner protein.
FEATURES Location/Qualifiers
source 1..5726
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 19..322
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 323..526
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
5'UTR 616..655
/label=HSV TK 5'-UTR
/note="5' untranslated region from the herpes simplex virus
thymidine kinase gene"
CDS 668..709
/label=myr
/note="N-myristoylation signal from Src kinase (Pellman et
al., 1985; Kaplan et al., 1988)"
CDS 716..1036
/label=DmrB
/note="F36V mutant of FK506-binding protein FKBP12"
CDS 1043..1363
/codon_start=1
/product="F36V mutant of FK506-binding protein FKBP12"
/label=F36V mutant of FK506-binding protein FKBP12
/note="DmrB"
/note="binds synthetic ligands that do not bind wild-type
FKBP"
/translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
KLE"
CDS 1370..1396
/label=HA
/note="HA (human influenza hemagglutinin) epitope tag"
intron 1428..2000
/label=beta-globin intron
/note="intron from rabbit beta-globin gene"
polyA_signal 2197..2252
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
rep_origin complement(2605..2740)
/direction=LEFT
/label=SV40 ori
/note="SV40 origin of replication"
rep_origin complement(3017..3605)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3776..4648)
/codon_start=1
/product="beta-lactamase"
/label=beta-lactamase
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFQVEVAMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDL
VEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTR
LDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAE
IGASLIKHW"
promoter complement(4649..4753)
/label=AmpR promoter
rep_origin complement(5085..5540)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 5685..5701
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 5708..5726
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"