Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V012406 | pcDNA4TO-mito-mCherry-24xGCN4_v1 | 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:
- pcDNA4TO-mito-mCherry-24xGCN4_v1
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8025 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Copy Number:
- High Copy
- Promoter:
- SV40
pcDNA4TO-mito-mCherry-24xGCN4_v1 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.
pcDNA4TO-mito-mCherry-24xGCN4_v1 vector Sequence
LOCUS 40924_10241 8025 bp DNA circular SYN 13-MAY-2021
DEFINITION A fusion protein of the SunTag 24xGCN4 v1 peptide array to a
mitochondrial targeting domain and mCherry.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8025)
AUTHORS Tanenbaum ME, Gilbert LA, Qi LS, Weissman JS, Vale RD
TITLE A Protein-Tagging System for Signal Amplification in Gene Expression
and Fluorescence Imaging.
JOURNAL Cell. 2014 Oct 8. pii: S0092-8674(14)01227-6. doi:
10.1016/j.cell.2014.09.039.
PUBMED 25307933
REFERENCE 2 (bases 1 to 8025)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 8025)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2014
Oct 8. pii: S0092-8674(14)01227-6. doi: 10.1016/j.cell.2014.09.039."
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8025
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 719..769
/label=CMVd2 promoter
/note="truncated human cytomegalovirus (CMV) immediate
early promoter that yields moderately reduced expression
relative to the full-length promoter"
protein_bind 771..789
/label=tet operator
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 792..810
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind 820..844
/label=LNCX
/note="Human CMV promoter, forward primer"
CDS 1163..1867
/codon_start=1
/label=mCherry
/note="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
GRHSTGGMDELYK"
CDS 1955..2020
/codon_start=1
/product="GCN4 peptide that allows binding to an scFv-GFP
fusion protein (Tanenbaum et al., 2014)"
/label=GCN4_v1
/translation="LLPKNYHLENEVARLKKLVGER"
CDS 2036..2101
/codon_start=1
/product="GCN4 peptide that allows binding to an scFv-GFP
fusion protein (Tanenbaum et al., 2014)"
/label=GCN4_v1
/translation="LLPKNYHLENEVARLKKLVGER"
CDS 2117..2182
/codon_start=1
/product="GCN4 peptide that allows binding to an scFv-GFP
fusion protein (Tanenbaum et al., 2014)"
/label=GCN4_v1
/translation="LLPKNYHLENEVARLKKLVGER"
CDS 2198..2263
/codon_start=1
/product="GCN4 peptide that allows binding to an scFv-GFP
fusion protein (Tanenbaum et al., 2014)"
/label=GCN4_v1
/translation="LLPKNYHLENEVARLKKLVGER"
CDS 2279..2344
/codon_start=1
/product="GCN4 peptide that allows binding to an scFv-GFP
fusion protein (Tanenbaum et al., 2014)"
/label=GCN4_v1
/translation="LLPKNYHLENEVARLKKLVGER"
CDS 2360..2425
/codon_start=1
/product="GCN4 peptide that allows binding to an scFv-GFP
fusion protein (Tanenbaum et al., 2014)"
/label=GCN4_v1
/translation="LLPKNYHLENEVARLKKLVGER"
CDS 2441..2506
/codon_start=1
/product="GCN4 peptide that allows binding to an scFv-GFP
fusion protein (Tanenbaum et al., 2014)"
/label=GCN4_v1
/translation="LLPKNYHLENEVARLKKLVGER"
CDS 2522..2587
/codon_start=1
/product="GCN4 peptide that allows binding to an scFv-GFP
fusion protein (Tanenbaum et al., 2014)"
/label=GCN4_v1
/translation="LLPKNYHLENEVARLKKLVGER"
CDS 2603..2668
/codon_start=1
/product="GCN4 peptide that allows binding to an scFv-GFP
fusion protein (Tanenbaum et al., 2014)"
/label=GCN4_v1
/translation="LLPKNYHLENEVARLKKLVGER"
CDS 2684..2749
/codon_start=1
/product="GCN4 peptide that allows binding to an scFv-GFP
fusion protein (Tanenbaum et al., 2014)"
/label=GCN4_v1
/translation="LLPKNYHLENEVARLKKLVGER"
CDS 2765..2830
/codon_start=1
/product="GCN4 peptide that allows binding to an scFv-GFP
fusion protein (Tanenbaum et al., 2014)"
/label=GCN4_v1
/translation="LLPKNYHLENEVARLKKLVGER"
CDS 2846..2911
/codon_start=1
/product="GCN4 peptide that allows binding to an scFv-GFP
fusion protein (Tanenbaum et al., 2014)"
/label=GCN4_v1
/translation="LLPKNYHLENEVARLKKLVGER"
primer_bind 2927..2949
/label=pGEX 5'
/note="pGEX vectors, Glutathione-S-transferase, forward
primer"
primer_bind 2950..2972
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS 2975..3040
/codon_start=1
/product="GCN4 peptide that allows binding to an scFv-GFP
fusion protein (Tanenbaum et al., 2014)"
/label=GCN4_v1
/translation="LLPKNYHLENEVARLKKLVGER"
CDS 3056..3121
/codon_start=1
/product="GCN4 peptide that allows binding to an scFv-GFP
fusion protein (Tanenbaum et al., 2014)"
/label=GCN4_v1
/translation="LLPKNYHLENEVARLKKLVGER"
CDS 3137..3202
/codon_start=1
/product="GCN4 peptide that allows binding to an scFv-GFP
fusion protein (Tanenbaum et al., 2014)"
/label=GCN4_v1
/translation="LLPKNYHLENEVARLKKLVGER"
CDS 3218..3283
/codon_start=1
/product="GCN4 peptide that allows binding to an scFv-GFP
fusion protein (Tanenbaum et al., 2014)"
/label=GCN4_v1
/translation="LLPKNYHLENEVARLKKLVGER"
CDS 3299..3364
/codon_start=1
/product="GCN4 peptide that allows binding to an scFv-GFP
fusion protein (Tanenbaum et al., 2014)"
/label=GCN4_v1
/translation="LLPKNYHLENEVARLKKLVGER"
CDS 3380..3445
/codon_start=1
/product="GCN4 peptide that allows binding to an scFv-GFP
fusion protein (Tanenbaum et al., 2014)"
/label=GCN4_v1
/translation="LLPKNYHLENEVARLKKLVGER"
CDS 3461..3526
/codon_start=1
/product="GCN4 peptide that allows binding to an scFv-GFP
fusion protein (Tanenbaum et al., 2014)"
/label=GCN4_v1
/translation="LLPKNYHLENEVARLKKLVGER"
CDS 3542..3607
/codon_start=1
/product="GCN4 peptide that allows binding to an scFv-GFP
fusion protein (Tanenbaum et al., 2014)"
/label=GCN4_v1
/translation="LLPKNYHLENEVARLKKLVGER"
CDS 3623..3688
/codon_start=1
/product="GCN4 peptide that allows binding to an scFv-GFP
fusion protein (Tanenbaum et al., 2014)"
/label=GCN4_v1
/translation="LLPKNYHLENEVARLKKLVGER"
CDS 3704..3769
/codon_start=1
/product="GCN4 peptide that allows binding to an scFv-GFP
fusion protein (Tanenbaum et al., 2014)"
/label=GCN4_v1
/translation="LLPKNYHLENEVARLKKLVGER"
CDS 3785..3850
/codon_start=1
/product="GCN4 peptide that allows binding to an scFv-GFP
fusion protein (Tanenbaum et al., 2014)"
/label=GCN4_v1
/translation="LLPKNYHLENEVARLKKLVGER"
CDS 3866..3931
/codon_start=1
/product="GCN4 peptide that allows binding to an scFv-GFP
fusion protein (Tanenbaum et al., 2014)"
/label=GCN4_v1
/translation="LLPKNYHLENEVARLKKLVGER"
polyA_signal 4042..4266
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 4312..4740
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 4754..5083
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
promoter 5131..5178
/label=EM7 promoter
/note="synthetic bacterial promoter"
CDS 5197..5568
/codon_start=1
/label=BleoR
/note="antibiotic-binding protein"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
polyA_signal 5701..5834
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5871..5887)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(5895..5911)
/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)."
promoter complement(5919..5949)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(5964..5985)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6102..6119)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6273..6858)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7032..7889)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(7890..7994)
/label=AmpR promoter