Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V000087 | pRK5-EGFP-Tau | 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:
- pRK5-EGFP-Tau
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6519 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Promoter:
- SV40
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- SP6
- 3' Primer:
- SV40pA-R
pRK5-EGFP-Tau vector 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.
pRK5-EGFP-Tau vector Sequence
LOCUS V000087 6519 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V000087
VERSION V000087
KEYWORDS pRK5-EGFP-Tau
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 6519)
AUTHORS Hoover BR, Reed MN, Su J, Penrod RD, Kotilinek LA, Grant MK,
Pitstick R, Carlson GA, Lanier LM, Yuan LL, Ashe KH, Liao D
TITLE Tau mislocalization to dendritic spines mediates synaptic
dysfunction independently of neurodegeneration.
JOURNAL Neuron. 2010 Dec 22;68(6):1067-81. doi:
10.1016/j.neuron.2010.11.030.
PUBMED 21172610
REFERENCE 2 (bases 1 to 6519)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 6519)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.neuron.2010.11"; journalName: "Neuron"; date:
"2010-12-22- 22"; volume: "68"; issue: "6"; pages: "1067-81"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6519
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 94..111
/label="L4440"
/note="L4440 vector, forward primer"
protein_bind 228..249
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 264..294
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 302..318
/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 326..342
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
enhancer 371..750
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 751..954
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 951..975
/label="LNCX"
/note="Human CMV promoter, forward primer"
promoter 1185..1203
/label="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 1216..1235
/label="pMT2-F"
/note="Synthetic intron, forward primer"
CDS 1285..2001
/label="EGFP"
/note="enhanced GFP"
CDS 2008..3153
/gene="MAPT"
/label="Microtubule-associated protein tau"
/note="Microtubule-associated protein tau from Papio
hamadryas. Accession#: Q9MYX8"
regulatory 3176..3185
/label="Kozak sequence"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
polyA_signal 3189..3323
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
promoter 3392..3749
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
primer_bind complement(3769..3785)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 3998..4453
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4470..4489)
/label="pRS-marker"
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4589..4611
/label="pGEX 3'"
/note="pGEX vectors, reverse primer"
primer_bind complement(4649..4667)
/label="pBRforEco"
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4735..4839
/label="AmpR promoter"
CDS 4840..5697
/label="AmpR"
/note="beta-lactamase"
rep_origin 5871..6459
/direction=RIGHT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"