Basic Vector Information
- Vector Name:
- pLenti-EF1α-DsRed-Monomer-T2A-Puro-U6-gRNA
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9532 bp
- Type:
- Gene knockout
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Puro
- Promoter:
- EF-1α
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pLenti-EF1α-DsRed-Monomer-T2A-Puro-U6-gRNA 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 ℃.
pLenti-EF1α-DsRed-Monomer-T2A-Puro-U6-gRNA vector Sequence
LOCUS 62056_13835 9532 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 9532)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..9532
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 1..634
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 681..806
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1303..1536
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1721..1765
/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 1914..1955
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
misc_feature 2027..2144
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2338..3519
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
CDS 3556..4230
/label=DsRed-Monomer
/note="monomeric derivative of DsRed fluorescent protein
(Strongin et al., 2007)"
CDS 4297..4893
/label=PuroR
/note="puromycin N-acetyltransferase"
polyA_signal 4897..5104
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
promoter 5112..5352
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
misc_RNA 5377..5452
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
misc_feature 5453..6041
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 6248..6881
/label=5' LTR
/note="5' long terminal repeat (LTR) from HIV-1"
primer_bind complement(7010..7026)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(7034..7050)
/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(7058..7088)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(7103..7124)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(7412..8000)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(8174..9031)
/label=AmpR
/note="beta-lactamase"
promoter complement(9032..9136)
/label=AmpR promoter
polyA_signal 9184..9318
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
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