Basic Vector Information
- Vector Name:
- Lenti-AsCpf1-Blast
- Antibiotic Resistance:
- Ampicillin
- Length:
- 12670 bp
- Type:
- Gene knockout
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Blast
- Promoter:
- EF1a
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
Lenti-AsCpf1-Blast 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 ℃.
Lenti-AsCpf1-Blast vector Sequence
LOCUS V016867 12670 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V016867
VERSION V016867
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 12670)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..12670
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 238..617
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 618..820
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 835..1015
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 1062..1187
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1680..1913
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 2098..2142
/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 2291..2332
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 2440..2557
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2650..2861
/label="EF-1-alpha core promoter"
/note="core promoter for human elongation factor
EF-1-alpha"
CDS 2903..6823
/label="AsCpf1"
/note="CRISPR-associated protein from Acidaminococcus sp.
BV3L6 (Zetsche et al., 2015)"
CDS 6881..6937
/label="P2A"
/note="2A peptide from porcine teschovirus-1 polyprotein"
CDS 6938..7333
/label="BSD"
/note="blasticidin S deaminase"
misc_feature 7361..7949
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
polyA_signal 8686..8910
/label="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin 8956..9384
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 9398..9728
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
promoter 9776..9823
/label="EM7 promoter"
/note="synthetic bacterial promoter"
CDS 9842..10213
/label="BleoR"
/note="antibiotic-binding protein"
polyA_signal 10346..10479
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
promoter complement(10564..10594)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(10609..10630)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(10918..11506)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(11680..12537)
/label="AmpR"
/note="beta-lactamase"
promoter complement(12538..12642)
/label="AmpR promoter"
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