Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V000601 | pcDNA4/TO/Myc-His /LacZ | 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
A Tetracycline-Regulated Expression System without Viral Transactivators The T-REx System yields higher levels of induced expression than any other regulated mammalian expression system. It utilizes the complete CMV promoter and adds control elements from the bacterial tetracycline resistance operon to effectively repress and derepress transcription from one of the strongest mammalian promoter sequences known (1,2).Specific Activation The T-REx System uses a repressor mechanism that blocks transcription from the powerful CMV promoter in the absence of tetracycline. Because the T-REx System elements do not use viral transactivators, you can achieve high-level expression from the complete CMV promoter without secondary, non-specific activation of host genes.The T-REx Mechanism The T-REx transcriptional control elements are illustrated in Figure 1. Two tetracycline operator sequences (TetO2) have been inserted between the TATA box of the CMV promoter and the
- Vector Name:
- pcDNA4/TO/Myc-His /LacZ
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8198 bp
- Type:
- Tetracycline Regulatory System
- Replication origin:
- ori
- Selection Marker:
- Zeocin
- Promoter:
- EM7
- Cloning Method:
- Enzyme digestion and ligation
- Fusion Tag:
- His Tag (6x), c-Myc Epitope Tag
pcDNA4/TO/Myc-His /LacZ 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.
pcDNA4/TO/Myc-His /LacZ vector Sequence
LOCUS 40924_10231 8198 bp DNA circular SYN 13-JAN-2022
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8198)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 8198)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8198
/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"
protein_bind 820..838
/label=tet operator
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 841..859
/gene="tetO"
/label=tetracycline repressor TetR binding site
/bound_moiety="tetracycline repressor TetR"
/note="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
CDS 1051..4095
/label=lacZ
/note="beta-galactosidase"
CDS 4120..4149
/label=Myc
/note="Myc (human c-Myc proto-oncogene) epitope tag"
CDS 4165..4182
/label=6xHis
/note="6xHis affinity tag"
polyA_signal 4211..4435
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 4481..4909
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 4923..5253
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
promoter 5301..5348
/label=EM7 promoter
/note="synthetic bacterial promoter"
CDS 5367..5738
/label=BleoR
/note="antibiotic-binding protein"
polyA_signal 5871..6004
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(6041..6057)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(6065..6081)
/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(6089..6119)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(6134..6155)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(6443..7031)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7205..8062)
/label=AmpR
/note="beta-lactamase"
promoter complement(8063..8167)
/label=AmpR promoter