Price Information
Cat No. | Plasmid Name | Availability | Add to cart |
---|---|---|---|
V014799 | pHR_Gal4UAS_EGFP_PGK_mCherry_Zeo | 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
- Vector Name:
- pHR_Gal4UAS_EGFP_PGK_mCherry_Zeo
- Antibiotic Resistance:
- Ampicillin
- Length:
- 11217 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Zeo
- Promoter:
- mPGK
- Growth Temperature:
- 37℃
pHR_Gal4UAS_EGFP_PGK_mCherry_Zeo 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.
pHR_Gal4UAS_EGFP_PGK_mCherry_Zeo vector Sequence
LOCUS V014799 11217 bp DNA circular SYN 01-JAN-1980 DEFINITION Exported. ACCESSION V014799 VERSION V014799 KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct . REFERENCE 1 (bases 1 to 11217) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..11217 /mol_type="other DNA" /organism="synthetic DNA construct" intron 455..520 /label="small t intron" /note="SV40 (simian virus 40) small t antigen intron" CDS 650..670 /label="SV40 NLS" /note="nuclear localization signal of SV40 (simian virus 40) large T antigen" polyA_signal 1095..1229 /label="SV40 poly(A) signal" /note="SV40 polyadenylation signal" LTR 1397..2030 /label="3' LTR" /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 2077..2202 /label="HIV-1 Psi" /note="packaging signal of human immunodeficiency virus type 1" misc_feature 2699..2932 /label="RRE" /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 3117..3161 /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 3310..3351 /note="Protein Tat from Human immunodeficiency virus type 1 group M subtype B (isolate WMJ22). Accession#: P12509" /label="Protein Tat" misc_feature 3446..3563 /label="cPPT/CTS" /note="central polypurine tract and central termination sequence of HIV-1" CDS 3894..4610 /label="EGFP" /note="enhanced GFP" promoter 4736..5235 /label="PGK promoter" /note="mouse phosphoglycerate kinase 1 promoter" CDS 5256..5963 /label="mCherry" /note="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" CDS 6027..6398 /label="BleoR" /note="antibiotic-binding protein" misc_feature 6477..7065 /label="WPRE" /note="woodchuck hepatitis virus posttranscriptional regulatory element" LTR 7175..7408 /label="3' LTR (Delta-U3)" /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" promoter complement(7433..7451) /label="SP6 promoter" /note="promoter for bacteriophage SP6 RNA polymerase" promoter 8239..8343 /label="AmpR promoter" CDS 8344..9201 /label="AmpR" /note="beta-lactamase" rep_origin 9375..9963 /direction=RIGHT /label="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" promoter 10209..10538 /label="SV40 promoter" /note="SV40 enhancer and early promoter"