- 载体名称:
- hCas9
- 载体抗性:
- Ampicillin
- 载体长度:
- 9553 bp
- 载体类型:
- CRISPR Plasmids
- 复制子:
- ori
- 载体来源:
- Mali P, Yang L, Esvelt KM, Aach J, Guell M, DiCarlo JE, Norville JE,
- 筛选标记:
- Neomycin/G418(Geneticin)
- 拷贝数:
- High copy number
- 启动子:
- SV40
hCas9 载体载体图谱
hCas9 载体载体序列
LOCUS hCas9. 9553 bp DNA circular SYN 01-JAN-1980 DEFINITION Mammalian vector with a G418 resistance marker for expressing human codon-optimized Cas9. ACCESSION . VERSION . KEYWORDS hCas9. SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9553) AUTHORS Mali P, Yang L, Esvelt KM, Aach J, Guell M, DiCarlo JE, Norville JE, Church GM. TITLE RNA-guided human genome engineering via Cas9. JOURNAL Science 2013;339:823-6. PUBMED 23287722 REFERENCE 2 (bases 1 to 9553) AUTHORS Church Lab / Addgene #41815 TITLE Direct Submission REFERENCE 3 (bases 1 to 9553) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Science"; date: "2013"; volume: "339"; pages: "823-6" COMMENT SGRef: number: 2; type: "Journal Article" COMMENT Minor sequence errors were corrected using the Addgene sequencing results. FEATURES Location/Qualifiers source 1..9553 /mol_type="other DNA" /organism="synthetic DNA construct" enhancer 50..429 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 430..633 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" CDS 748..4851 /label=Cas9 /note="Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" CDS 4864..4884 /codon_start=1 /product="nuclear localization signal of SV40 large T antigen" /note="SV40 NLS" /translation="PKKKRKV" polyA_signal 4983..5031 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5233..5661 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 5675..6004 /label=SV40 promoter /note="SV40 enhancer and early promoter" CDS 6071..6862 /label=NeoR/KanR /note="aminoglycoside phosphotransferase" polyA_signal 7041..7174 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(7211..7227) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(7235..7251) /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(7259..7289) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(7304..7325) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(7613..8201) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(8375..9232) /label=AmpR /note="beta-lactamase" promoter complement(9233..9337) /label=AmpR promoter
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