Basic Vector Information
- Vector Name:
- pLV-Tet-miniCMV-MCS-GFP-nanobody-mCherry-Puro
- Antibiotic Resistance:
- Ampicillin
- Length:
- 11146 bp
- Type:
- Protein expression, Tetracycline inducible
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Puro
- Promoter:
- mPGK
- 5' Primer:
- miniCMV-F
- Growth Temperature:
- 37℃
pLV-Tet-miniCMV-MCS-GFP-nanobody-mCherry-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 ℃.
pLV-Tet-miniCMV-MCS-GFP-nanobody-mCherry-Puro vector Sequence
LOCUS V015072 11146 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V015072
VERSION V015072
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 11146)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..11146
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 25..142
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
protein_bind 210..228
/label="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
promoter 418..456
/label="minimal CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 524..542
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 603..629
/label="HA"
/note="HA (human influenza hemagglutinin) epitope tag"
CDS 675..1016
/label="GFP nanobody"
/note="GFP-binding fragment of a single-chain camelid
antibody (Rothbauer et al., 2008)"
promoter 1142..1641
/label="PGK promoter"
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 1662..2369
/label="mCherry"
/note="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
CDS 2427..3023
/label="PuroR"
/note="puromycin N-acetyltransferase"
misc_feature 3037..3625
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 3688..3921
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
promoter complement(3946..3964)
/label="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
promoter 4753..4857
/label="AmpR promoter"
CDS 4858..5715
/label="AmpR"
/note="beta-lactamase"
rep_origin 5889..6477
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 6723..7052
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
intron 8186..8251
/label="small t intron"
/note="SV40 (simian virus 40) small t antigen intron"
CDS 8381..8401
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
polyA_signal 8826..8960
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
LTR 9129..9762
/label="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 9809..9934
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 10426..10659
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 10844..10888
/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 11037..11078
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
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