Basic Vector Information
- Vector Name:
- pLV2-CMV-MCS-3×FLAG-IRES-EGFP-PGK-Puro
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9448 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Puro
- Promoter:
- mPGK
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pLV2-CMV-MCS-3×FLAG-IRES-EGFP-PGK-Puro 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 ℃.
pLV2-CMV-MCS-3×FLAG-IRES-EGFP-PGK-Puro vector Sequence
LOCUS V014257 9448 bp DNA circular SYN 01-JAN-1980 DEFINITION Exported. ACCESSION V014257 VERSION V014257 KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct . REFERENCE 1 (bases 1 to 9448) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..9448 /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" /label="Protein Tat" misc_feature 2027..2144 /label="cPPT/CTS" /note="central polypurine tract and central termination sequence of HIV-1" enhancer 2201..2504 /label="CMV enhancer" /note="human cytomegalovirus immediate early enhancer" promoter 2505..2708 /label="CMV promoter" /note="human cytomegalovirus (CMV) immediate early promoter" CDS 2837..2902 /label="3xFLAG" /note="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" misc_feature 2912..3498 /label="IRES2" /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" CDS 3500..4216 /label="EGFP" /note="enhanced GFP" promoter 4236..4735 /label="PGK promoter" /note="mouse phosphoglycerate kinase 1 promoter" CDS 4756..5352 /label="PuroR" /note="puromycin N-acetyltransferase" misc_feature 5369..5957 /label="WPRE" /note="woodchuck hepatitis virus posttranscriptional regulatory element" LTR 6164..6797 /label="5' LTR" /note="5' long terminal repeat (LTR) from HIV-1" primer_bind complement(6926..6942) /label="M13 rev" /note="common sequencing primer, one of multiple similar variants" protein_bind complement(6950..6966) /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(6974..7004) /label="lac promoter" /note="promoter for the E. coli lac operon" protein_bind complement(7019..7040) /label="CAP binding site" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(7328..7916) /direction=LEFT /label="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(8090..8947) /label="AmpR" /note="beta-lactamase" promoter complement(8948..9052) /label="AmpR promoter" polyA_signal 9100..9234 /label="SV40 poly(A) signal" /note="SV40 polyadenylation signal"
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