Basic Vector Information
- Vector Name:
- pLV-sgRNA
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7465 bp
- Type:
- Genome editing
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Blast
- Promoter:
- hPGK
- 5' Primer:
- U6
- Growth Temperature:
- 37℃
pLV-sgRNA 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-sgRNA vector Sequence
LOCUS 62056_14600 7465 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 7465) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..7465 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 5..231 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" misc_feature 459..584 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1077..1310 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1495..1539 /codon_start=1 /label=gp41 peptide /note="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /translation="KNEQELLELDKWASL" promoter 1714..2214 /label=hPGK promoter /note="human phosphoglycerate kinase 1 promoter" CDS 2226..2621 /codon_start=1 /label=BSD /note="blasticidin S deaminase" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" misc_feature 2681..2798 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2916..3156 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" misc_RNA 3173..3248 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" misc_feature 3322..3910 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" LTR 3982..4215 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 4293..4427 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin 4454..4589 /label=SV40 ori /note="SV40 origin of replication" promoter complement(4610..4628) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(4638..4654) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" rep_origin 4796..5251 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 5277..5381 /label=AmpR promoter CDS 5382..6239 /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 6413..7001 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" protein_bind 7289..7310 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 7325..7355 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7363..7379 /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)." primer_bind 7387..7403 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" promoter 7424..7442 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase"
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