Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V000678 | FUW mCherry-GFP-LC3 | In stock (lyophilized plasmid) |
Buy one, get one free! |
Two vials of lyophilized plasmid will be delivered, each vial is about 5µg.
Basic Vector Information
The FUW mCherry-GFP-LC3 contains GFP and mCherry fluorescence. It can to observe free autophagosomes and autophagosomes fused to lysosomes (autolysosomes; mCherry fluorescence only because GFP is acid sensitive).
- Vector Name:
- FUW mCherry-GFP-LC3
- Antibiotic Resistance:
- Ampicillin
- Length:
- 11138 bp
- Type:
- Lentiviral
- Replication origin:
- ori
- Promoter:
- UbC
- Cloning Method:
- TOPO Cloning
- 5' Primer:
- CGCGGATCCGGTCGCCACCATGGTGAGCAAGGGCGAG
- 3' Primer:
- CGCGGCGCGCCGCTGGGTCTAGATGCATGC
- Growth Strain(s):
- stbl3
- Growth Temperature:
- 37℃
FUW mCherry-GFP-LC3 vector Vector Map
Plasmid Resuspension 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 ℃.
References
- Leeman DS, Hebestreit K, Ruetz T, Webb AE, McKay A, Pollina EA, Dulken BW, Zhao X, Yeo RW, Ho TT, Mahmoudi S, Devarajan K, Passegué E, Rando TA, Frydman J, Brunet A. Lysosome activation clears aggregates and enhances quiescent neural stem cell activation during aging. Science. 2018 Mar 16;359(6381):1277-1283.
FUW mCherry-GFP-LC3 vector Sequence
LOCUS FUW_mCherry-GFP- 11138 bp DNA circular SYN 13-MAY-2021
DEFINITION To visualize free autophagosomes (GFP and mCherry fluorescence) and
autophagosomes that have fused with the lysosome (autolyosomes;
mCherry fluorescence only, due to acid sensitivity of GFP).
ACCESSION .
VERSION .
KEYWORDS FUW mCherry-GFP-LC3.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11138)
AUTHORS Leeman DS, Hebestreit K, Ruetz T, Webb AE, McKay A, Pollina EA,
Dulken BW, Zhao X, Yeo RW, Ho TT, Mahmoudi S, Devarajan K, Passegue
E, Rando TA, Frydman J, Brunet A
TITLE Lysosome activation clears aggregates and enhances quiescent neural
stem cell activation during aging.
JOURNAL Science. 2018 Mar 16;359(6381):1277-1283. doi:
10.1126/science.aag3048. Epub 2018 Mar 15.
PUBMED 29590078
REFERENCE 2 (bases 1 to 11138)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 11138)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1126/science.aag3048"; journalName: "Science"; date:
"2018-03-16- 16"; volume: "359"; issue: "6381"; pages: "1277-1283"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..11138
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 7..386
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 387..589
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 604..784
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 831..956
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1449..1682
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1867..1911
/label=gp41 peptide
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"
CDS 2060..2101
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
misc_feature 2209..2326
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2391..3602
/label=UbC promoter
/note="human ubiquitin C promoter"
CDS 3647..4354
/label=mCherry
/note="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
protein_bind 4382..4406
/label=attB1
/note="recombination site for the Gateway(R) BP reaction"
CDS 4421..5137
/label=EGFP
/note="enhanced GFP"
CDS 5165..5539
/codon_start=1
/gene="MAP1LC3B"
/product="human LC3 protein"
/label=LC3B
/note="Atg8 family member found on autophagosomes"
/translation="MPSEKTFKQRRTFEQRVEDVRLIREQHPTKIPVIIERYKGEKQLP
VLDKTKFLVPDHVNMSELIKIIRRRLQLNANQAFFLLVNGHSMVSVSTPISEVYESEKD
EDGFLYMVYASQETFGMKLSV"
misc_feature 5599..6187
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(6190..6206)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6191..6207)
/label=pBluescriptKS
/note="For pBluescript vector"
LTR 6712..6892
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
polyA_signal 6924..7148
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 7194..7622
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 7636..7965
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
promoter 8013..8060
/label=EM7 promoter
/note="synthetic bacterial promoter"
CDS 8079..8450
/label=BleoR
/note="antibiotic-binding protein"
polyA_signal 8583..8716
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(8753..8769)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8753..8769)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(8766..8788)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 8777..8793
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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)."
promoter complement(8801..8831)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(8846..8867)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(8984..9001)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(9155..9743)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(9917..10774)
/label=AmpR
/note="beta-lactamase"
promoter complement(10775..10879)
/label=AmpR promoter
primer_bind complement(10954..10973)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"