载体基本信息 | |||
载体名称 | JDS246 | 载体抗性 | Ampicillin |
---|---|---|---|
载体长度 | 7614 bp | 载体类型 | CRISPR Plasmids |
载体来源 | Joung Lab / Addgene #43861 | 拷贝数 | High copy number |
JDS246 载体载体图谱
JDS246 载体载体序列
LOCUS JDS246. 7614 bp DNA circular SYN 01-JAN-1980 DEFINITION Plasmid for expressing FLAG(R)-tagged human codon-optimized Cas9 in mammalian cells. ACCESSION . VERSION . KEYWORDS JDS246. SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7614) AUTHORS Joung Lab / Addgene #43861 TITLE Direct Submission REFERENCE 2 (bases 1 to 7614) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..7614 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 132..335 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" promoter 377..395 /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" CDS 409..4512 /label=Cas9 /note="Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" CDS 4519..4539 /codon_start=1 /product="nuclear localization signal of SV40 large T antigen" /note="SV40 NLS" /translation="PKKKRKV" CDS 4546..4611 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=three tandem FLAG /note="3xFLAG" /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS 4629..4646 /label=6xHis /note="6xHis affinity tag" polyA_signal 4675..4899 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(4970..4986) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(4994..5010) /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)." promoter complement(5018..5048) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(5063..5084) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(5372..5960) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6134..6991) /label=AmpR /note="beta-lactamase" promoter complement(6992..7096) /label=AmpR promoter enhancer 7366..7614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer"
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