Price Information
Cat No. | Plasmid Name | Availability | Add to cart |
---|---|---|---|
V017172 | pLenti-DsRed_IRES_SNCA:EGFP | 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:
- pLenti-DsRed_IRES_SNCA:EGFP
- Antibiotic Resistance:
- Ampicillin
- Length:
- 10354 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Neo/G418
- Promoter:
- CMV
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pLenti-DsRed_IRES_SNCA:EGFP 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.
pLenti-DsRed_IRES_SNCA:EGFP vector Sequence
LOCUS V017172 10354 bp DNA circular SYN 01-JAN-1980 DEFINITION Exported. ACCESSION V017172 VERSION V017172 KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct . REFERENCE 1 (bases 1 to 10354) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..10354 /mol_type="other DNA" /organism="synthetic DNA construct" LTR 376..633 /label="5' LTR (truncated)" /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 680..805 /label="HIV-1 Psi" /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1302..1535 /label="RRE" /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1720..1764 /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 1913..1954 /note="Protein Tat from Human immunodeficiency virus type 1 group M subtype B (isolate WMJ22). Accession#: P12509" /label="Protein Tat" misc_feature 2057..2174 /label="cPPT/CTS" /note="central polypurine tract and central termination sequence of HIV-1" enhancer 2226..2529 /label="CMV enhancer" /note="human cytomegalovirus immediate early enhancer" promoter 2530..2733 /label="CMV promoter" /note="human cytomegalovirus (CMV) immediate early promoter" CDS 2843..3517 /label="DsRed-Express" /note="rapidly maturing tetrameric variant of DsRed fluorescent protein (Bevis and Glick, 2002)" misc_feature 3546..4130 /label="IRES2" /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" CDS 4571..5287 /label="EGFP" /note="enhanced GFP" promoter complement(5311..5329) /label="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" promoter 5339..5814 /label="PGK promoter" /note="mouse phosphoglycerate kinase 1 promoter" CDS 5835..6626 /label="NeoR/KanR" /note="aminoglycoside phosphotransferase" misc_feature 6743..7331 /label="WPRE" /note="woodchuck hepatitis virus posttranscriptional regulatory element" LTR 7406..7639 /label="3' LTR (Delta-U3)" /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" promoter 7844..7948 /label="AmpR promoter" CDS 7949..8806 /label="AmpR" /note="beta-lactamase" rep_origin 8980..9568 /label="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" protein_bind 9979..10000 /label="CAP binding site" /note="CAP binding activates transcription in the presence of cAMP." LTR 10000..10354 /label="3' LTR" /note="3' long terminal repeat (LTR) from HIV-1"