Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V000877 | pCS2 Notch1 Full Length-6MT | 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:
- pCS2 Notch1 Full Length-6MT
- Antibiotic Resistance:
- Ampicillin
- Length:
- 11042 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Copy Number:
- High Copy
- Promoter:
- sCMV
- 5' Primer:
- Sp6
- 3' Primer:
- EBV-rev
pCS2 Notch1 Full Length-6MT 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.
pCS2 Notch1 Full Length-6MT vector Sequence
LOCUS 40924_13660 11042 bp DNA circular SYN 13-MAY-2021
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11042)
AUTHORS Schroeter EH, Kisslinger JA, Kopan R
TITLE Notch-1 signalling requires ligand-induced proteolytic release of
intracellular domain.
JOURNAL Nature. 1998 May 28;393(6683):382-6.
PUBMED 9620803
REFERENCE 2 (bases 1 to 11042)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 11042)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nature.";
date: "1998-05-28"; volume: "393(6683)"; pages: "382-6"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..11042
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter complement(21..999)
/label=CMV IE94 promoter
/note="enhancer/promoter region of simian cytomegalovirus
major immediate early transcription unit IE94"
rep_origin 1076..1531
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 1557..1661
/label=AmpR promoter
CDS 1662..2519
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 2693..3281
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3435..3452
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 3569..3590
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3605..3635
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3643..3659
/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)."
primer_bind 3648..3670
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 3667..3683
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 3667..3683
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
promoter 3704..3722
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
polyA_signal 3835..3969
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
promoter 3974..3991
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS complement(4019..4048)
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS complement(4082..4111)
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS complement(4121..4150)
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS complement(4160..4189)
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS complement(4199..4228)
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS complement(4238..4267)
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
primer_bind complement(4281..4297)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4282..4298)
/label=pBluescriptKS
/note="For pBluescript vector"
regulatory 9018..9027
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
regulatory 10287..10296
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
regulatory 10619..10628
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
promoter complement(11010..11028)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"