Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V013312 | pCDH-EF1-MCS-PGK-Puro | In stock, instant shipping |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
pCDH-EF1-MCS-PGK-Puro enables stable expression of the target gene driven by the EF1 promoter, with the PGK promoter regulating puromycin resistance screening. This facilitates stable cell line construction and gene function studies.
- Vector Name:
- pCDH-EF1-MCS-PGK-Puro
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7540 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Puro
- Promoter:
- mPGK
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pCDH-EF1-MCS-PGK-Puro 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
- Yang L, Chen Y, Liu N, Lu Y, Li X, Ma W, Gan W, Li D. 5mC and H3K9me3 of TRAF3IP2 promoter region accelerates the progression of translocation renal cell carcinoma. Biomark Res. 2022 Jul 27;10(1):54. doi: 10.1186/s40364-022-00402-3. PMID: 35897085; PMCID: PMC9331078.
pCDH-EF1-MCS-PGK-Puro vector Sequence
LOCUS Exported 7540 bp DNA circular SYN 10-NOV-2025
DEFINITION Exported.
ACCESSION V013312
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7540)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 7540)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7540
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 6..232
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 233..413
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 457..582
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1075..1308
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1492..1536
/label=gp41 peptide
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"
CDS 1685..1726
/label=Protein Tat
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
misc_feature 1797..1914
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 1959..2170
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
LTR 2183..2451
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from human
T-cell leukemia virus (HTLV) type 1"
promoter 2509..3008
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 3029..3625
/label=PuroR
/note="puromycin N-acetyltransferase"
misc_feature 3635..4223
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 4297..4530
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 4602..4736
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin 4742..4877
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(4915..4931)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(4939..4955)
/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(4963..4993)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(5008..5029)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5317..5905)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6079..6936)
/label=AmpR
/note="beta-lactamase"
promoter complement(6937..7041)
/label=AmpR promoter
primer_bind 7515..7531
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"