Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V000806 | pcDNA4TO-24xGCN4_v4-kif18b-24xPP7 | 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-24xGCN4_v4-kif18b-24xPP7
- Antibiotic Resistance:
- Ampicillin
- Length:
- 10960 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Selection Marker:
- Zeocin
- Copy Number:
- High Copy
pcDNA4TO-24xGCN4_v4-kif18b-24xPP7 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-24xGCN4_v4-kif18b-24xPP7 vector Sequence
LOCUS 40924_10236 10960 bp DNA circular SYN 13-MAY-2021
DEFINITION Translation reporter.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10960)
AUTHORS Yan X, Hoek TA, Vale RD, Tanenbaum ME
TITLE Dynamics of Translation of Single mRNA Molecules In Vivo.
JOURNAL Cell. 2016 May 5;165(4):976-89. doi: 10.1016/j.cell.2016.04.034.
PUBMED 27153498
REFERENCE 2 (bases 1 to 10960)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 10960)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.cell.2016.04"; journalName: "Cell"; date: "2016-05-5- 5";
volume: "165"; issue: "4"; pages: "976-89"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..10960
/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 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
protein_bind 820..838
/label=tet operator
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 841..859
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind 869..893
/label=LNCX
/note="Human CMV promoter, forward primer"
regulatory 984..993
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 1059..1115
/codon_start=1
/label=GCN4_v4
/note="GCN4 peptide optimized to improve solubility while
preserving binding to an scFv-GFP fusion protein (Tanenbaum
et al., 2014)"
/translation="EELLSKNYHLENEVARLKK"
CDS 1131..1187
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 1203..1259
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 1275..1331
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 1347..1403
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 1419..1475
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 1491..1547
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 1563..1619
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 1635..1691
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 1707..1763
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 1779..1835
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 1851..1907
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 1923..1979
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 1995..2051
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 2067..2123
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 2139..2195
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 2211..2267
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 2283..2339
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 2355..2411
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 2427..2483
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 2499..2555
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 2571..2627
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 2715..2771
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
polyA_signal 6977..7201
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 7247..7675
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 7689..8018
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
promoter 8066..8113
/label=EM7 promoter
/note="synthetic bacterial promoter"
CDS 8132..8503
/codon_start=1
/label=BleoR
/note="antibiotic-binding protein"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
polyA_signal 8636..8769
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(8806..8822)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8806..8822)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(8819..8841)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 8830..8846
/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(8854..8884)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(8899..8920)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(9037..9054)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(9208..9793)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(9967..10824)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(10825..10929)
/label=AmpR promoter