Price Information
Cat No. | Plasmid Name | Availability | Add to cart |
---|---|---|---|
V014580 | pAAV-hSyn-eNpHR3.0-mCherry | 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:
- pAAV-hSyn-eNpHR3.0-mCherry
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6239 bp
- Type:
- Gene expression
- Replication origin:
- ori
- Host:
- Mammalian cells, Adeno-associated virus
- Promoter:
- SYN1
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pAAV-hSyn-eNpHR3.0-mCherry 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.
pAAV-hSyn-eNpHR3.0-mCherry vector Sequence
LOCUS V014580 6239 bp DNA circular SYN 01-JAN-1980 DEFINITION Exported. ACCESSION V014580 VERSION V014580 KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct . REFERENCE 1 (bases 1 to 6239) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..6239 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 226..673 /label="hSyn promoter" /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" CDS 713..1585 /gene="hop" /label="Halorhodopsin" /note="Halorhodopsin from Natronomonas pharaonis. Accession#: P15647" CDS 1655..2362 /label="mCherry" /note="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" misc_feature 2417..3005 /label="WPRE" /note="woodchuck hepatitis virus posttranscriptional regulatory element" polyA_signal 3037..3513 /label="hGH poly(A) signal" /note="human growth hormone polyadenylation signal" repeat_region 3553..3693 /label="AAV2 ITR" /note="inverted terminal repeat of adeno-associated virus serotype 2" rep_origin 3768..4223 /label="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 4505..4609 /label="AmpR promoter" CDS 4610..5467 /label="AmpR" /note="beta-lactamase" rep_origin 5641..6229 /direction=RIGHT /label="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"