Basic Vector Information
- Vector Name:
- pLV3-CMV-EGFP-TurboID-NLS-3×FLAG-Puro
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9731 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Puro
- Promoter:
- mPGK
- 5' Primer:
- CMV-F
- Growth Temperature:
- 37℃
pLV3-CMV-EGFP-TurboID-NLS-3×FLAG-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-EGFP-TurboID-NLS-3×FLAG-Puro vector Sequence
LOCUS V014823 9731 bp DNA circular SYN 01-JAN-1980 DEFINITION Exported. ACCESSION V014823 VERSION V014823 KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct . REFERENCE 1 (bases 1 to 9731) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..9731 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 27..131 /label="AmpR promoter" CDS 132..989 /label="AmpR" /note="beta-lactamase" rep_origin 1163..1751 /label="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" protein_bind 2039..2060 /label="CAP binding site" /note="CAP binding activates transcription in the presence of cAMP." promoter 2075..2105 /label="lac promoter" /note="promoter for the E. coli lac operon" protein_bind 2113..2129 /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)." primer_bind 2137..2153 /label="M13 rev" /note="common sequencing primer, one of multiple similar variants" promoter 2174..2192 /label="T3 promoter" /note="promoter for bacteriophage T3 RNA polymerase" promoter 2220..2447 /label="RSV promoter" /note="Rous sarcoma virus enhancer/promoter" LTR 2448..2628 /label="5' LTR (truncated)" /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 2672..2797 /label="HIV-1 Psi" /note="packaging signal of human immunodeficiency virus type 1" misc_feature 3290..3523 /label="RRE" /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 3707..3751 /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 3900..3941 /note="Protein Tat from Human immunodeficiency virus type 1 group M subtype B (isolate WMJ22). Accession#: P12509" /label="Protein Tat" misc_feature 4012..4129 /label="cPPT/CTS" /note="central polypurine tract and central termination sequence of HIV-1" enhancer 4152..4455 /label="CMV enhancer" /note="human cytomegalovirus immediate early enhancer" promoter 4456..4659 /label="CMV promoter" /note="human cytomegalovirus (CMV) immediate early promoter" CDS 4765..5481 /label="EGFP" /note="enhanced GFP" CDS 5554..6510 /label="TurboID" /note="mutant of the E. coli biotin ligase BioID engineered for improved catalytic efficiency (Branon et al., 2018)" CDS 6517..6537 /label="SV40 NLS" /note="nuclear localization signal of SV40 (simian virus 40) large T antigen" CDS 6580..6603 /label="FLAG" /note="FLAG(R) epitope tag, followed by an enterokinase cleavage site" misc_feature 6613..7201 /label="WPRE" /note="woodchuck hepatitis virus posttranscriptional regulatory element" promoter 7215..7714 /label="PGK promoter" /note="mouse phosphoglycerate kinase 1 promoter" CDS 7735..8331 /label="PuroR" /note="puromycin N-acetyltransferase" LTR 8472..8705 /label="3' LTR (Delta-U3)" /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 8777..8911 /label="SV40 poly(A) signal" /note="SV40 polyadenylation signal" rep_origin 8917..9052 /label="SV40 ori" /note="SV40 origin of replication" promoter complement(9085..9103) /label="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(9113..9129) /label="M13 fwd" /note="common sequencing primer, one of multiple similar variants" rep_origin 9274..9729 /direction=RIGHT /label="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis"
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