Basic Vector Information
- Vector Name:
- CKS2 gRNA (BRDN0001146116)
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8323 bp
- Type:
- Gene knockout
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Puro
- Promoter:
- EF-1α
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
CKS2 gRNA (BRDN0001146116) 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 ℃.
CKS2 gRNA (BRDN0001146116) vector Sequence
LOCUS 62056_526 8323 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 8323) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..8323 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 87..1265 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" CDS 1278..1874 /codon_start=1 /label=PuroR /note="puromycin N-acetyltransferase" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" misc_feature 1893..2481 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" LTR 2553..2786 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 2864..2998 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin 3025..3160 /label=SV40 ori /note="SV40 origin of replication" promoter complement(3181..3199) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(3209..3225) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" rep_origin 3367..3822 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 3848..3952 /label=AmpR promoter CDS 3953..4810 /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 4984..5572 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" protein_bind 5860..5881 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 5896..5926 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5934..5950 /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 5958..5974 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" promoter 5995..6013 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 6041..6267 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 6268..6448 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 6495..6620 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 7113..7346 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 7531..7575 /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 7753..7993 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" misc_RNA 8023..8098 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" misc_feature 8154..8271 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1"
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