Basic Vector Information
C-term SpCas9 piece of inducible split-5 (Cas9(C)-FKBP split-5).
PX854 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 ℃.
PX854 vector Sequence
LOCUS PX854. 7087 bp DNA circular SYN 11-SEP-2021 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS PX854. SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7087) TITLE Direct Submission REFERENCE 2 (bases 1 to 7087) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..7087 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 1..241 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" misc_RNA 268..343 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" enhancer 440..725 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer; contains an 18-bp deletion relative to the standard CMV enhancer" promoter 727..1004 /label=chicken beta-actin promoter intron 1005..1233 /note="hybrid intron" /note="hybrid between chicken beta-actin (CBA) and minute virus of mice (MMV) introns (Gray et al., 2011)" CDS 1257..1277 /label=SV40 NLS /note="nuclear localization signal of SV40 (simian virus 40) large T antigen" CDS 1302..1622 /label=FKBP /note="human FK506-binding protein FKBP12" CDS 1668..4052 /label=Cas9(C) /note="C-terminal portion of Streptococcus pyogenes Cas9 (Zetsche et al., 2015)" CDS 4053..4100 /label=nucleoplasmin NLS /note="bipartite nuclear localization signal from nucleoplasmin" polyA_signal 4134..4341 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" repeat_region 4350..4490 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" rep_origin 4565..5020 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 5302..5406 /label=AmpR promoter CDS 5407..6264 /label=AmpR /note="beta-lactamase" rep_origin 6438..7026 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
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