Basic Vector Information
- Vector Name:
- pMLLV-EF1a-Fluc-IRES-EGFP-PGK-Puro-WPRE
- Antibiotic Resistance:
- Ampicillin
- Length:
- 11866 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Puro
- Promoter:
- EF-1α
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pMLLV-EF1a-Fluc-IRES-EGFP-PGK-Puro-WPRE vector Vector Map
Plasmid Resuspension 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 ℃.
pMLLV-EF1a-Fluc-IRES-EGFP-PGK-Puro-WPRE vector Sequence
LOCUS 62056_17150 11866 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11866) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..11866 /mol_type="other DNA" /organism="synthetic DNA construct" LTR 1..634 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 681..806 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1303..1536 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1721..1765 /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 1914..1955 /note="Protein Tat from Human immunodeficiency virus type 1 group M subtype B (isolate WMJ22). Accession#: P12509" misc_feature 2027..2144 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2338..3519 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" CDS 3535..5184 /label=luciferase /note="firefly luciferase" misc_feature 5214..5788 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" CDS 5818..6534 /label=EGFP /note="enhanced GFP" promoter 6567..7065 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" CDS 7125..7721 /label=PuroR /note="puromycin N-acetyltransferase" misc_feature 7787..8375 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" LTR 8582..9215 /label=5' LTR /note="5' long terminal repeat (LTR) from HIV-1" promoter complement(9392..9422) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(9437..9458) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(9746..10334) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(10508..11365) /label=AmpR /note="beta-lactamase" promoter complement(11366..11470) /label=AmpR promoter polyA_signal 11518..11652 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal"
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