Basic Vector Information
- Vector Name:
- pLV3-attB-EF1a-lncRNA-CMV-Fluc-T2A-Puro
- Antibiotic Resistance:
- Ampicillin
- Length:
- 10709 bp
- Type:
- Luciferase reporter
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Puro
- Promoter:
- EF-1α
- Growth Temperature:
- 37℃
pLV3-attB-EF1a-lncRNA-CMV-Fluc-T2A-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-attB-EF1a-lncRNA-CMV-Fluc-T2A-Puro vector Sequence
LOCUS V014475 10709 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V014475
VERSION V014475
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 10709)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..10709
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 237..616
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 617..819
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
misc_feature 1061..1186
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1683..1916
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 2101..2145
/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 2294..2335
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 2443..2560
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
protein_bind 2619..2639
/label="attB4"
/note="core recombination site for the Gateway(R) BP
reaction"
promoter 2640..3818
/label="EF-1-alpha promoter"
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
protein_bind 3818..3842
/label="attB1"
/note="recombination site for the Gateway(R) BP reaction"
protein_bind complement(4390..4414)
/label="attB2"
/note="recombination site for the Gateway(R) BP reaction"
promoter 4817..5016
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 5045..5063
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 5084..6733
/label="luciferase"
/note="firefly luciferase"
CDS 6803..6856
/label="T2A"
/note="2A peptide from Thosea asigna virus capsid protein"
CDS 6857..7453
/label="PuroR"
/note="puromycin N-acetyltransferase"
protein_bind complement(7509..7542)
/label="loxP"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
misc_feature 7598..8186
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 8715..8895
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
rep_origin complement(8957..9545)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(9719..10576)
/label="AmpR"
/note="beta-lactamase"
promoter complement(10577..10681)
/label="AmpR promoter"
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