Basic Vector Information
- Vector Name:
- Lenti sgRNA(MS2)_zeo backbone
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9997 bp
- Type:
- Gene knockout
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Zeo
- Promoter:
- EF-1α
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
Lenti sgRNA(MS2)_zeo backbone 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 sgRNA(MS2)_zeo backbone vector Sequence
LOCUS V016597 9997 bp DNA circular SYN 01-JAN-1980 DEFINITION Exported. ACCESSION V016597 VERSION V016597 KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct . REFERENCE 1 (bases 1 to 9997) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..9997 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 18..347 /label="SV40 promoter" /note="SV40 enhancer and early promoter" promoter 395..442 /label="EM7 promoter" /note="synthetic bacterial promoter" CDS 461..832 /label="BleoR" /note="antibiotic-binding protein" polyA_signal 965..1098 /label="SV40 poly(A) signal" /note="SV40 polyadenylation signal" promoter complement(1183..1213) /label="lac promoter" /note="promoter for the E. coli lac operon" protein_bind complement(1228..1249) /label="CAP binding site" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(1537..2125) /direction=LEFT /label="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(2299..3156) /label="AmpR" /note="beta-lactamase" promoter complement(3157..3261) /label="AmpR promoter" enhancer 3527..3906 /label="CMV enhancer" /note="human cytomegalovirus immediate early enhancer" promoter 3907..4109 /label="CMV promoter" /note="human cytomegalovirus (CMV) immediate early promoter" LTR 4124..4304 /label="5' LTR (truncated)" /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 4351..4476 /label="HIV-1 Psi" /note="packaging signal of human immunodeficiency virus type 1" misc_feature 4969..5202 /label="RRE" /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 5387..5431 /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 5580..5621 /note="Protein Tat from Human immunodeficiency virus type 1 group M subtype B (isolate WMJ22). Accession#: P12509" /label="Protein Tat" misc_feature 5729..5846 /label="cPPT/CTS" /note="central polypurine tract and central termination sequence of HIV-1" promoter 5896..6136 /label="U6 promoter" /note="RNA polymerase III promoter for human U6 snRNA" misc_RNA 6182..6200 /label="MS2 stem loop" /note="stem loop that binds the bacteriophage MS2 coat protein" promoter 6394..7572 /label="EF-1-alpha promoter" /note="strong constitutive promoter for human elongation factor EF-1-alpha" CDS 7579..7950 /label="BleoR" /note="antibiotic-binding protein" misc_feature 7978..8566 /label="WPRE" /note="woodchuck hepatitis virus posttranscriptional regulatory element" polyA_signal 9303..9527 /label="bGH poly(A) signal" /note="bovine growth hormone polyadenylation signal" rep_origin 9573..9997 /label="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis"
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