Basic Vector Information
- Vector Name:
- pLV-PGK-dAsCas12f1-VPR-P65-RtaAD-mCherry
- Antibiotic Resistance:
- Ampicillin
- Length:
- 12722 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Promoter:
- mPGK
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pLV-PGK-dAsCas12f1-VPR-P65-RtaAD-mCherry 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-PGK-dAsCas12f1-VPR-P65-RtaAD-mCherry vector Sequence
LOCUS V014331 12722 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V014331
VERSION V014331
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 12722)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..12722
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 27..144
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 209..708
/label="PGK promoter"
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 780..800
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
CDS 813..2078
/gene="cas12f1"
/label="CRISPR-associated endodeoxyribonuclease Cas12f1"
/note="CRISPR-associated endodeoxyribonuclease Cas12f1 from
Sulfoacidibacillus thermotolerans. Accession#: A0A2U3D0N8"
CDS 2100..2126
/label="HA"
/note="HA (human influenza hemagglutinin) epitope tag"
CDS 2766..3137
/label="RelA (p65) AD"
/note="transcriptional activation domain of human RelA,
also known as p65 (O'Shea and Perkins, 2008)"
CDS 3156..3725
/label="Rta AD"
/note="transcriptional activation domain from the human
herpesvirus 4 (Epstein-Barr virus) replication and
transcription activator Rta/BRLF1 (Hardwick et al., 1992;
Chavez et al., 2015)"
CDS 3753..3779
/label="c-myc NLS"
/note="nuclear localization signal of human c-Myc
proto-oncogene (Dang and Lee, 1988)"
CDS 3792..4496
/label="mCherry"
/note="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
misc_feature 4563..5151
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 5261..5494
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
promoter complement(5519..5537)
/label="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
promoter 6325..6429
/label="AmpR promoter"
CDS 6430..7287
/label="AmpR"
/note="beta-lactamase"
rep_origin 7461..8049
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 8295..8624
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
intron 9758..9823
/label="small t intron"
/note="SV40 (simian virus 40) small t antigen intron"
CDS 9953..9973
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
polyA_signal 10398..10532
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
LTR 10700..11333
/label="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 11380..11505
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 12002..12235
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 12420..12464
/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 12613..12654
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
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