Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V012181 | pVQ CMV NanoV1-2a-EGFP ferritin | 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:
- pVQ CMV NanoV1-2a-EGFP ferritin
- Antibiotic Resistance:
- Ampicillin
- Length:
- 11340 bp
- Type:
- Mammalian Expression, Adenoviral
- Replication origin:
- ori
- Promoter:
- CMV
- 5' Primer:
- GACTACAAGGACGACGACGA
pVQ CMV NanoV1-2a-EGFP ferritin 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.
pVQ CMV NanoV1-2a-EGFP ferritin vector Sequence
LOCUS V012181 11340 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V012181
VERSION V012181
KEYWORDS pVQ CMV NanoV1-2a-EGFP ferritin
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 11340)
AUTHORS Stanley SA, Kelly L, Latcha KN, Schmidt SF, Yu X, Nectow AR, Sauer
J, Dyke JP, Dordick JS, Friedman JM
TITLE Bidirectional electromagnetic control of the hypothalamus regulates
feeding and metabolism.
JOURNAL Nature. 2016 Mar 31;531(7596):647-50. doi: 10.1038/nature17183. Epub
2016 Mar 23.
PUBMED 27007848
REFERENCE 2 (bases 1 to 11340)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 11340)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1038/nature17183"; journalName: "Nature"; date: "2016-03-31-
31"; volume: "531"; issue: "7596"; pages: "647-50"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..11340
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 389..411
/label="pGEX 3'"
/note="pGEX vectors, reverse primer"
polyA_signal 554..688
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
CDS complement(1328..1351)
/label="FLAG"
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
RBS 1506..1514
/label="Shine-Dalgarno sequence"
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
CDS complement(1928..2644)
/label="EGFP"
/note="enhanced GFP"
CDS complement(2660..2713)
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label="T2A"
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
CDS complement(2714..5227)
/gene="Trpv1"
/label="Transient receptor potential cation channel
subfamily V member 1"
/note="Transient receptor potential cation channel
subfamily V member 1 from Rattus norvegicus. Accession#:
O35433"
CDS complement(5258..5599)
/codon_start=1
/product="GFP-binding fragment of a single-chain camelid
antibody (Rothbauer et al., 2008)"
/label="GFP nanobody"
/translation="VQLVESGGALVQPGGSLRLSCAASGFPVNRYSMRWYRQAPGKERE
WVAGMSSAGDRSSYEDSVKGRFTISRDDARNTVYLQMNSLKPEDTAVYYSNVNVGFEYW
GQGTQVTVSS"
regulatory 5605..5614
/label="Kozak sequence"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
protein_bind complement(5643..5667)
/label="attB1"
/note="recombination site for the Gateway(R) BP reaction"
protein_bind complement(5694..5727)
/label="loxP"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
promoter complement(5775..5978)
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
enhancer complement(5979..6282)
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
protein_bind complement(6372..6392)
/label="attB4"
/note="core recombination site for the Gateway(R) BP
reaction"
primer_bind complement(6418..6434)
/label="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6419..6435)
/label="pBluescriptKS"
/note="For pBluescript vector"
misc_signal complement(6476..6626)
/label="Ad5 Psi"
/note="packaging signal for adenovirus serotype 5"
repeat_region 6714..6816
/label="ITR"
/note="inverted terminal repeat of human adenovirus
serotype 5"
primer_bind complement(6844..6862)
/label="pBRforEco"
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 6930..7034
/label="AmpR promoter"
CDS 7035..7892
/label="AmpR"
/note="beta-lactamase"
rep_origin 8066..8654
/direction=RIGHT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS 9507..10841
/gene="IVa2"
/label="Packaging protein 1"
/note="Packaging protein 1 from Human adenovirus C serotype
5. Accession#: P03271"
CDS complement(10907..11326)
/gene="IX"
/label="Hexon-interlacing protein"
/note="Hexon-interlacing protein from Human adenovirus C
serotype 5. Accession#: P03281"