Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V003417 | pSCCA5-E8d | 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:
- pSCCA5-E8d
- Antibiotic Resistance:
- Ampicillin
- Length:
- 4575 bp
- Type:
- Expression vector
- Replication origin:
- ori
- Source/Author:
- Arimitsu H, Sasaki K, Iba Y, Kurosawa Y, Shimizu T, Tsuji T.
pSCCA5-E8d 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.
pSCCA5-E8d vector Sequence
LOCUS 40924_38963 4575 bp DNA circular SYN 18-DEC-2018
DEFINITION Expression vector pSCCA5-E8d DNA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4575)
AUTHORS Arimitsu H, Sasaki K, Iba Y, Kurosawa Y, Shimizu T, Tsuji T.
TITLE Isolation of B subunit-specific monoclonal antibody clones that
strongly neutralize the toxicity of Shiga toxin 2
JOURNAL Microbiol. Immunol. 59 (2), 71-81 (2015)
PUBMED 25521016
REFERENCE 2 (bases 1 to 4575)
AUTHORS Arimitsu H, Iba Y.
TITLE Direct Submission
JOURNAL Submitted (06-JUN-2014) Contact:Hideyuki Arimitsu Fujita Health
University, Department of Microbiology; 1-98, Dengakugakubo,
Kutsukake-cho, Toyoake, Aichi 470-1192, Japan URL
:https://fujita-hu.ac.jp/
REFERENCE 3 (bases 1 to 4575)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 4575)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Microbiol.
Immunol."; date: "2015"; volume: "59"; issue: "2"; pages: "71-81"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(06-JUN-2014) Contact:Hideyuki Arimitsu Fujita Health University,
Department of Microbiology"; volume: " 1-98, Dengakugakubo,
Kutsukake-cho, Toyoake, Aichi 470-1192, Japan URL :https"; pages:
"//fujita-hu.ac.jp"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4575
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 40..87
/label=PelB leader sequence
/note="PelB leader sequence"
misc_feature 197..241
/label=Single chain linker
/note="Single chain linker"
CDS 375..692
/codon_start=1
/label=mIg-kappa-CL
/note="Mouse immunoglobulin kappa light chain constant
region"
/translation="ADAAPTVSIFPPSSEQLTSGGASVVCFLNSFYPKDINVKWKIDGS
ERQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRN
EC"
misc_feature 711..1340
/label=cp3
/note="cp3"
CDS 1393..1566
/codon_start=1
/label=ProtA
/note="IgG-binding unit of Staphylococcus aureus protein A"
/translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
AEAKKLNDAQAPK"
CDS 1570..1740
/codon_start=1
/product="IgG-binding unit of Staphylococcus aureus protein
A"
/label=ProtA
/translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
EAKKLNDAQAPK"
primer_bind complement(1770..1786)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin complement(1928..2383)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2410..2514
/label=AmpR promoter
CDS 2515..3372
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 3546..4134
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 4422..4443
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4458..4488
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4496..4512
/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)."
primer_bind 4520..4536
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 4555..4573
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"