Basic Vector Information
- Vector Name:
- Lenti-Cas9-Puro
- Antibiotic Resistance:
- Ampicillin
- Length:
- 12700 bp
- Type:
- Gene knockout
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Puro,Zeo
- Promoter:
- SV40
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
Lenti-Cas9-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 ℃.
Lenti-Cas9-Puro vector Sequence
LOCUS V013322 12700 bp DNA circular SYN 01-JAN-1980 DEFINITION Exported. ACCESSION V013322 VERSION V013322 KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct . REFERENCE 1 (bases 1 to 12700) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..12700 /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" 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 2702..2913 /label="EF-1-alpha core promoter" /note="core promoter for human elongation factor EF-1-alpha" CDS 2938..7041 /label="Cas9" /note="Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" CDS 7090..7113 /label="FLAG" /note="FLAG(R) epitope tag, followed by an enterokinase cleavage site" CDS 7123..7179 /label="P2A" /note="2A peptide from porcine teschovirus-1 polyprotein" CDS 7180..7773 /label="PuroR" /note="puromycin N-acetyltransferase" misc_feature 7792..8380 /label="WPRE" /note="woodchuck hepatitis virus posttranscriptional regulatory element" LTR 8452..8685 /label="3' LTR (Delta-U3)" /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 8717..8941 /label="bGH poly(A) signal" /note="bovine growth hormone polyadenylation signal" rep_origin 8987..9415 /label="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 9429..9758 /label="SV40 promoter" /note="SV40 enhancer and early promoter" promoter 9806..9853 /label="EM7 promoter" /note="synthetic bacterial promoter" CDS 9872..10243 /label="BleoR" /note="antibiotic-binding protein" polyA_signal 10376..10509 /label="SV40 poly(A) signal" /note="SV40 polyadenylation signal" promoter complement(10594..10624) /label="lac promoter" /note="promoter for the E. coli lac operon" protein_bind complement(10639..10660) /label="CAP binding site" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(10948..11536) /direction=LEFT /label="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(11710..12567) /label="AmpR" /note="beta-lactamase" promoter complement(12568..12672) /label="AmpR promoter"
This page is informational only.