Basic Vector Information
- Vector Name:
- pLV3-CMV-3×FLAG-MCS-TurboID-HA-Puro
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8515 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Puro
- Promoter:
- RSV
- 5' Primer:
- CMV-F
- Growth Temperature:
- 37℃
pLV3-CMV-3×FLAG-MCS-TurboID-HA-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 ℃.
pLV3-CMV-3×FLAG-MCS-TurboID-HA-Puro vector Sequence
LOCUS V014926 8515 bp DNA circular SYN 01-JAN-1980 DEFINITION Exported. ACCESSION V014926 VERSION V014926 KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct . REFERENCE 1 (bases 1 to 8515) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..8515 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 6..233 /label="RSV promoter" /note="Rous sarcoma virus enhancer/promoter" LTR 234..414 /label="5' LTR (truncated)" /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 458..583 /label="HIV-1 Psi" /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1076..1309 /label="RRE" /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1493..1537 /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 1686..1727 /note="Protein Tat from Human immunodeficiency virus type 1 group M subtype B (isolate WMJ22). Accession#: P12509" /label="Protein Tat" misc_feature 1804..1920 /label="cPPT/CTS" /note="central polypurine tract and central termination sequence of HIV-1" promoter 2010..2213 /label="CMV promoter" /note="human cytomegalovirus (CMV) immediate early promoter" CDS 2293..2364 /label="3xFLAG" /note="three tandem FLAG(R) epitope tags" CDS 2389..3348 /label="TurboID" /note="mutant of the E. coli biotin ligase BioID engineered for improved catalytic efficiency (Branon et al., 2018)" CDS 3352..3378 /label="HA" /note="HA (human influenza hemagglutinin) epitope tag" misc_feature 3397..3974 /label="IRES2" /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" CDS 4004..4600 /label="PuroR" /note="puromycin N-acetyltransferase" misc_feature 4610..5198 /label="WPRE" /note="woodchuck hepatitis virus posttranscriptional regulatory element" LTR 5272..5505 /label="3' LTR (Delta-U3)" /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 5577..5711 /label="SV40 poly(A) signal" /note="SV40 polyadenylation signal" rep_origin 5717..5852 /label="SV40 ori" /note="SV40 origin of replication" primer_bind complement(5890..5906) /label="M13 rev" /note="common sequencing primer, one of multiple similar variants" protein_bind complement(5914..5930) /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(5938..5968) /label="lac promoter" /note="promoter for the E. coli lac operon" protein_bind complement(5983..6004) /label="CAP binding site" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(6292..6880) /direction=LEFT /label="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(7054..7911) /label="AmpR" /note="beta-lactamase" promoter complement(7912..8016) /label="AmpR promoter" primer_bind 8490..8506 /label="M13 fwd" /note="common sequencing primer, one of multiple similar variants"
This page is informational only.