Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V011584 | pTriEx-3 Neo | In stock, 1 week for quality controls |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pTriEx-3 Neo
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6445 bp
- Type:
- Insect Cell Vectors
- Replication origin:
- ori
- Source/Author:
- Novagen (EMD Millipore)
- Selection Marker:
- Neomycin/G418(Geneticin)
- Copy Number:
- High copy number
- Promoter:
- CMV
pTriEx-3 Neo 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.
pTriEx-3 Neo vector Sequence
LOCUS pTriEx-3_Neo. 6445 bp DNA circular SYN 01-JAN-1980
DEFINITION Insect, bacterial, and mammalian vector with a CMV promoter and a
neomycin resistance marker, for expressing proteins with a
C-terminal HSV-8xHis cassette.
ACCESSION .
VERSION .
KEYWORDS pTriEx-3 Neo
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6445)
AUTHORS Novagen (EMD Millipore)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 6445)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6445
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_recomb 181..1008
/label=baculovirus recombination region (lef2/ORF603)
/note="contains ORF603 and part of lef2"
enhancer 1076..1379
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 1381..1580
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
exon 1597..1698
/note="Exon 1"
intron 1699..1862
promoter 1931..1949
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 1950..1974
/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 1990..2099
/label=p10 promoter
/note="baculovirus promoter for expression in insect cells"
RBS 2101..2106
/note="ribosome binding site"
misc_feature 2112..2219
/label=MCS
/note="MCS"
/note="multiple cloning site"
CDS 2114..2116
/codon_start=1
/product="start codon"
/label=start codon
/note="ATG"
/translation="M"
CDS 2225..2257
/label=HSV tag
/note="HSV (herpes simplex virus) epitope tag"
CDS 2264..2287
/label=8xHis
/note="8xHis affinity tag"
misc_feature 2386..2848
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 2884..3684
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase from Tn5"
polyA_signal 3805..3860
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
terminator 3948..3995
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
misc_recomb 4008..4713
/label=baculovirus recombination region (ORF1629)
/note="contains part of ORF1629"
protein_bind complement(4722..4755)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
rep_origin complement(4923..5510)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5526..6383)
/label=AmpR
/note="beta-lactamase"