Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V010158 | pSecTag2A-E06-scFv | In stock, instant shipping |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
The E06 antibody is a murine monoclonal IgM natural antibody that binds to oxidized phospholipid and reacts with oxidized LDL, oxidized HDL and protein covalently modified by oxidized phospholipid. This antibody has been used to probe atherosclerotic plaque in mice, zebrafish, rabbits, monkeys and humans. It also binds to apoptotic cells, but not viable cells, and can be used in a variety of applications, including ELISA, immunohistochemistry and Western blot analysis, as well as for in vivo imaging with MRI techniques.
- Vector Name:
- pSecTag2A-E06-scFv
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5846 bp
- Type:
- Mammalian Expression Vectors
- Replication origin:
- ori
- Source/Author:
- Dr. Joseph Witztum
- Selection Marker:
- Zeocin
- Copy Number:
- High copy number
- Promoter:
- CMV
- Cloning Method:
- Enzyme Cut
- 5' Primer:
- T7 promoter
- 3' Primer:
- BGH Reverse primer
- Fusion Tag:
- Myc,6*His
- Growth Strain(s):
- JM108
- Growth Temperature:
- 37℃
pSecTag2A-E06-scFv 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.
References
- Que X, Hung MY, Yeang C, Gonen A, Prohaska TA, Sun X, Diehl C, Määttä A, Gaddis DE, Bowden K, Pattison J, MacDonald JG, Ylä-Herttuala S, Mellon PL, Hedrick CC, Ley K, Miller YI, Glass CK, Peterson KL, Binder CJ, Tsimikas S, Witztum JL. Oxidized phospholipids are proinflammatory and proatherogenic in hypercholesterolaemic mice. Nature. 2018 Jun;558(7709):301-306.
pSecTag2A-E06-scFv vector Sequence
LOCUS Exported 5846 bp DNA circular SYN 02-SEP-2024
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS pSecTag2A-E06-scFv
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5846)
AUTHORS Triple Threat
TITLE Direct Submission
REFERENCE 2 (bases 1 to 5846)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 5846)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5846
/mol_type="other DNA"
/organism="synthetic DNA construct"
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"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
sig_peptide 905..967
/label=Ig-kappa leader
/note="leader sequence from mouse immunoglobulin kappa
light
chain"
CDS 968..1342
/codon_start=1
/label=E06 VL
/note="E06 VL"
/translation="AAQPARRAVRSLDIVMTQSPSSLSVSAGKKVTISCTASESLYSSK
HKVHYLAWYQKKPEQSPKLLIYGASNRYIGVPDRFTGSGSGTDFTLTISSVQVEDLTHY
YCAQFYSYPLTFGAGTKLEIK"
CDS 1343..1387
/codon_start=1
/label=G4S x 3
/note="G4S x 3"
/translation="GGGGSGGGGSGGGGS"
CDS 1388..1768
/codon_start=1
/label=E06 VH
/note="E06 VH"
/translation="EVKLVESGGGLVQPGGSLRLSCATSGFTFSDFYMEWVRQAPGKRL
EWIAASRNKANDYTTEYADSVKGRFIVSRDTSQSILYLQMNALRAEDTAIYYCARDYYG
SSYWYFDVWGAGTTVTVSSRGGP"
CDS 1769..1798
/codon_start=1
/product="Myc (human c-Myc oncogene) epitope tag"
/label=Myc
/note="Myc"
/translation="EQKLISEEDL"
CDS 1814..1831
/label=6xHis
/note="6xHis affinity tag"
polyA_signal 1860..2084
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2130..2558
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2572..2901
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
promoter 2949..2996
/label=EM7 promoter
/note="synthetic bacterial promoter"
CDS 3015..3386
/label=BleoR
/note="antibiotic-binding protein"
polyA_signal 3519..3652
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3689..3705)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(3713..3729)
/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(3737..3767)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3782..3803)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4091..4679)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4853..5710)
/label=AmpR
/note="beta-lactamase"
promoter complement(5711..5815)
/label=AmpR promoter