Basic Vector Information
Also known as pX601-AAV-CMV::NLS-SaCas9-NLS-3xHA-bGHpA;U6::BsaI-sgRNA
- Vector Name:
- pX601
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7447 bp
- Type:
- CRISPR/Cas
- Replication origin:
- ori
- Copy Number:
- High copy number
- Promoter:
- U6
- Fusion Tag:
- HA
pX601 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 ℃.
pX601 vector Sequence
LOCUS pX601. 7447 bp DNA circular SYN 11-SEP-2021 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS pX601. SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7447) TITLE Direct Submission REFERENCE 2 (bases 1 to 7447) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..7447 /mol_type="other DNA" /organism="synthetic DNA construct" enhancer 154..533 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 534..737 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" CDS 768..788 /codon_start=1 /product="nuclear localization signal of SV40 large T antigen" /label=nuclear localization signal of SV40 large T ant /note="SV40 NLS" /translation="PKKKRKV" CDS 813..3968 /label=SaCas9 /note="Cas9 endonuclease from the Staphylococcus aureus Type II CRISPR/Cas system" CDS 3969..4016 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=bipartite nuclear localization signal from nucl /note="nucleoplasmin NLS" /translation="KRPAATKKAGQAKKKK" CDS 4023..4103 /label=3xHA /note="three tandem HA epitope tags" polyA_signal 4137..4344 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" promoter 4352..4592 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" misc_RNA 4621..4696 /label=Sa gRNA scaffold /note="guide RNA scaffold for the Staphylococcus aureus CRISPR/Cas9 system" repeat_region 4710..4850 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" rep_origin 4925..5380 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 5662..5766 /label=AmpR promoter CDS 5767..6624 /label=AmpR /note="beta-lactamase" rep_origin 6798..7386 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
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