Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V001733 | DsRed-rab7 WT | 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:
- DsRed-rab7 WT
- Antibiotic Resistance:
- Kanamycin
- Length:
- 6060 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Selection Marker:
- Neomycin (select with G418)
- Copy Number:
- High Copy
- Promoter:
- CMV
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- DsRed1-C
- 3' Primer:
- SV40pA-R
DsRed-rab7 WT 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.
DsRed-rab7 WT vector Sequence
LOCUS V001733 6060 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V001733
VERSION V001733
KEYWORDS DsRed-rab7 WT
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 6060)
AUTHORS Choudhury A, Dominguez M, Puri V, Sharma DK, Narita K, Wheatley CL,
Marks DL, Pagano RE
TITLE Rab proteins mediate Golgi transport of caveola-internalized
glycosphingolipids and correct lipid trafficking in Niemann-Pick C
cells.
JOURNAL J Clin Invest. 2002 Jun . 109(12):1541-50.
PUBMED 12070301
REFERENCE 2 (bases 1 to 6060)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 6060)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Clin
Invest. 2002 Jun . 109(12):1541-50."
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6060
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 68..371
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 372..575
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 620..1297
/label="DsRed1"
/note="wild-type DsRed"
misc_feature 1301..1357
/label="MCS"
/note="multiple cloning site"
CDS 1361..1387
/label="HA"
/note="HA (human influenza hemagglutinin) epitope tag"
CDS 1388..1414
/label="HA"
/note="HA (human influenza hemagglutinin) epitope tag"
CDS 1418..2035
/gene="RAB7A"
/label="Ras-related protein Rab-7a"
/note="Ras-related protein Rab-7a from Homo sapiens.
Accession#: P51149"
CDS 2048..2074
/label="HA"
/note="HA (human influenza hemagglutinin) epitope tag"
CDS 2075..2101
/label="HA"
/note="HA (human influenza hemagglutinin) epitope tag"
CDS 2105..2722
/gene="RAB7A"
/label="Ras-related protein Rab-7a"
/note="Ras-related protein Rab-7a from Homo sapiens.
Accession#: P51149"
polyA_signal 2855..2976
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin complement(2983..3438)
/direction=LEFT
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 3465..3569
/label="AmpR promoter"
promoter 3571..3928
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
CDS 3963..4754
/label="NeoR/KanR"
/note="aminoglycoside phosphotransferase"
primer_bind complement(4945..4964)
/label="TK-pA-R"
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4989..5036
/label="HSV TK poly(A) signal"
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 5365..5953
/direction=RIGHT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"