Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V011590 | pTriEx-1.1 Neo | 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
- Vector Name:
- pTriEx-1.1 Neo
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6662 bp
- Type:
- Insect Cell Vectors
- Replication origin:
- ori
- Source/Author:
- Novagen (EMD Millipore)
- Selection Marker:
- Neomycin/G418(Geneticin)
- Copy Number:
- High copy number
- Promoter:
- chicken β-actin
- Growth Strain(s):
- DH10b
- Growth Temperature:
- 37℃
pTriEx-1.1 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-1.1 Neo vector Sequence
LOCUS Exported 6662 bp DNA circular SYN 03-SEP-2024
DEFINITION Insect, bacterial, and mammalian vector with a neomycin resistance
marker, for expressing proteins with a C-terminal HSV-8xHis
cassette.
ACCESSION .
VERSION .
KEYWORDS pTriEx-1.1 Neo
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6662)
AUTHORS Novagen (EMD Millipore)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 6662)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 6662)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6662
/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 1140..1443
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 1449..1724
/label=chicken beta-actin promoter
exon 1725..1817
/note="Exon 1"
intron 1818..2079
promoter 2148..2166
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 2167..2191
/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 2207..2316
/label=p10 promoter
/note="baculovirus promoter for expression in insect cells"
RBS 2318..2323
/note="ribosome binding site"
misc_feature 2329..2436
/label=MCS
/note="MCS"
/note="multiple cloning site"
CDS 2331..2333
/codon_start=1
/product="start codon"
/label=start codon
/note="ATG"
/translation="M"
CDS 2442..2474
/label=HSV tag
/note="HSV (herpes simplex virus) epitope tag"
CDS 2481..2504
/label=8xHis
/note="8xHis affinity tag"
misc_feature 2603..3065
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 3101..3901
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase from Tn5"
polyA_signal 4022..4077
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
terminator 4165..4212
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
misc_recomb 4225..4930
/label=baculovirus recombination region (ORF1629)
/note="contains part of ORF1629"
protein_bind complement(4939..4972)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
rep_origin complement(5140..5727)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5743..6600)
/label=AmpR
/note="beta-lactamase"