Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V016715 | GW1-PercevalHR | In stock, instant shipping |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- GW1-PercevalHR
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7557 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells
- Promoter:
- CMV
- Growth Strain(s):
- DH10B
- Growth Temperature:
- 37℃
GW1-PercevalHR 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.
GW1-PercevalHR vector Sequence
LOCUS Exported 7557 bp DNA circular SYN 28-MAR-2025
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS GW1-PercevalHR
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7557)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..7557
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 309..688
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 689..892
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
5'UTR 893..1872
/label=5'UTR of IE
/note="5'UTR of Human cytomegalovirus (HCMV) major
immediate-early protein (IE) gene"
CDS 2060..3799
/codon_start=1
/label=PercevalHR
/note="a fluorescent sensor of ATP-to-ADP ratio"
/translation="MKKVESIIRPEKLEIVKKALSDAGYVGMTVSEVKGSGVQGGIFER
YRGRVYSAGYNSDNVYITADKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVL
LPDNHYLSFQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGSGGMVSKGEELFT
GVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGL
QCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKG
IDFKEDGNILGHKLEYNGTIVDLIPKVKIELVVKEEDVDNVIDIICENARTGNPGDGKI
FVIPVERVVRVRTKEEGASGGGSGGGGASGMKKVEAIIRPEKLEIVKKALSDAGYVGMT
VSEVKGRGAGGGDLIPKVKIELVVKEEDVDNVIDIICENARTGNPGDGKIFVIPVERIV
RVRTKEEGASGGGGGSGGASGMKKVEAIIRPEKLEIVKKALNDDGYVGMTVSEVKGRGA
GGGDLIPKVKIELVVKEEDVDNIIDIICENARTGNPGDGKIFVIPVERVVRPRTKEEGK
EAL"
CDS 2252..2503
/codon_start=1
/product="C-terminal fragment of mVenus for use in
bimolecular fluorescence complementation (BiFC) (Kodama and
Hu, 2010)"
/label=VC155
/translation="DKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNH
YLSFQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
polyA_signal 3921..4055
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4078..4094)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 4102..4118
/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(4126..4156)
/label=lac promoter
/note="promoter for the E. coli lac operon"
rep_origin complement(4479..5067)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5238..6098)
/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"
rep_origin complement(6155..6668)
/direction=LEFT
/label=M13 ori
/note="M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 7284..7300
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"