Basic Vector Information
- Vector Name:
- pLV3-CMV-MCS-3×HA-CopGFP-Puro
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8325 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Puro
- Promoter:
- EF-1α core
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pLV3-CMV-MCS-3×HA-CopGFP-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-MCS-3×HA-CopGFP-Puro vector Sequence
LOCUS 62056_15200 8325 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 8325) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..8325 /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" misc_feature 1804..1921 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2011..2214 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" CDS 2333..2359 /label=HA /note="HA (human influenza hemagglutinin) epitope tag" CDS 2363..2389 /label=HA /note="HA (human influenza hemagglutinin) epitope tag" CDS 2393..2419 /label=HA /note="HA (human influenza hemagglutinin) epitope tag" promoter 2484..2695 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" LTR 2708..2976 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" CDS 3032..3697 /label=CopGFP /note="green fluorescent protein 2 from Pontellina plumata, also known as ppluGFP2 (Shagin et al., 2004)" CDS 3767..3820 /label=T2A /note="2A peptide from Thosea asigna virus capsid protein" CDS 3821..4417 /label=PuroR /note="puromycin N-acetyltransferase" misc_feature 4421..5008 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" LTR 5082..5315 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 5387..5521 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin 5527..5662 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(5700..5716) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(5724..5740) /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(5748..5778) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(5793..5814) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(6102..6690) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6864..7721) /label=AmpR /note="beta-lactamase" promoter complement(7722..7826) /label=AmpR promoter primer_bind 8300..8316 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants"
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