Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V000865 | pscAAV-CAG-GFP | 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
The pscAAV-CAG-GFP is a recombinant AAV vector packaging self-complementary GFP under the CAG promoter
- Vector Name:
- pscAAV-CAG-GFP
- Antibiotic Resistance:
- Ampicillin
- Length:
- 4801 bp
- Type:
- Mammalian Expression, AAV
- Replication origin:
- ori
- Copy Number:
- High Copy
- Promoter:
- chicken β-actin
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- GTTATTGTGCTGTCTCATC
- 3' Primer:
- CCACACCTCCCCCTGAAC
- Growth Strain(s):
- DH10b
- Growth Temperature:
- 37℃
pscAAV-CAG-GFP 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
- Pekrun K, De Alencastro G, Luo QJ, Liu J, Kim Y, Nygaard S, Galivo F, Zhang F, Song R, Tiffany MR, Xu J, Hebrok M, Grompe M, Kay MA. Using a barcoded AAV capsid library to select for clinically relevant gene therapy vectors. JCI Insight. 2019 Nov 14;4(22):e131610.
pscAAV-CAG-GFP vector Sequence
LOCUS 40924_38848 4801 bp DNA circular SYN 13-MAY-2021
DEFINITION recombinant AAV vector packaging self-complementary GFP under the
CAG promoter.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4801)
AUTHORS Pekrun K, De Alencastro G, Luo QJ, Liu J, Kim Y, Nygaard S, Galivo
F, Zhang F, Song R, Tiffany MR, Xu J, Hebrok M, Grompe M, Kay MA
TITLE Using a barcoded AAV capsid library to select for clinically
relevant gene therapy vectors.
JOURNAL JCI Insight. 2019 Nov 14;4(22). pii: 131610. doi:
10.1172/jci.insight.131610.
PUBMED 31723052
REFERENCE 2 (bases 1 to 4801)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 4801)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "JCI
Insight."; date: "2019-11-14"; pages: " 10.1172/jci.insight.131610"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4801
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 417..696
/label=chicken beta-actin promoter
primer_bind 975..997
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1059..1078
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
CDS 1124..1840
/codon_start=1
/label=EGFP
/note="enhanced GFP"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind 1887..1906
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(2156..2172)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2381..2400)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2500..2522
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(2560..2578)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 2646..2750
/label=AmpR promoter
CDS 2751..3608
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 3782..4370
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4524..4541
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 4658..4679
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4694..4724
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4732..4748
/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 4756..4772
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"