Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V006530 | pEPI-1 | 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 pEPI-1 vector contains two mammalian transcription units and a total of 305 CpG islands, which are located predominantly within the vector elements necessary for bacterial propagation and known to be counterproductive for persistent long-term transgene expression. The episomal replication of vector pEPI-1 depends on a transcription unit starting from the constitutively expressed Cytomegalovirus immediate early promoter (CMV-IEP) and directed into a 2000 bp long matrix attachment region sequence (MARS) derived from the human β-interferon gene.
- Vector Name:
- pEPI-1
- Antibiotic Resistance:
- Kanamycin
- Length:
- 6944 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Zhang X, Wang XY, Jia YL, Guo X, Wang YF, Wang TY.
- Growth Strain(s):
- DH10b
- Growth Temperature:
- 37℃
pEPI-1 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
- Piechaczek C, Fetzer C, Baiker A, Bode J, Lipps HJ. A vector based on the SV40 origin of replication and chromosomal S/MARs replicates episomally in CHO cells. Nucleic Acids Res. 1999;27(2):426-428. doi:10.1093/nar/27.2.426
- Wang XY, Zhang X, Wang TY, Jia YL, Xu DH, Yi DD. Shortened nuclear matrix attachment regions are sufficient for replication and maintenance of episomes in mammalian cells. Mol Biol Cell. 2019;30(22):2761-2770. doi:10.1091/mbc.E19-02-0108
pEPI-1 vector Sequence
LOCUS Exported 6944 bp DNA circular SYN 29-AUG-2024
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS pEPI-1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6944)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 6944)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6944
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 1..304
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 305..508
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 553..1269
/label=EGFP
/note="enhanced GFP"
misc_feature 1276..3476
/label=S/MAR
/note="S/MAR"
/note="S/MARs-element(Scaffold or Matrix Attachment
Regions) are typically 70% A/T-rich sequences and are often
found in association with chromosomal origins of
bidirectional replication. It could be used to construct
non-viral episomal vectors in mammalian cells."
misc_feature 3483..3548
/label=MCS
/note="multiple cloning site"
polyA_signal 3672..3793
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin complement(3800..4255)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 4282..4386
/label=AmpR promoter
promoter 4388..4745
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
CDS 4780..5571
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
polyA_signal 5806..5853
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 6182..6770
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 6885..6944
/label=CMV
/note="Human cytomegalovirus immediate early
enhancer/promoter"