hCas9 载体 (V012337#)

载体基本信息
载体名称 hCas9 载体抗性 Ampicillin
载体长度 9553 bp 载体类型 CRISPR Plasmids
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

hCas9 载体载体图谱

hCas99553 bp400800120016002000240028003200360040004400480052005600600064006800720076008000840088009200CMV enhancerCMV promoterCas9SV40 NLSHSV TK poly(A) signalf1 oriSV40 promoterNeoR/KanRSV40 poly(A) signalM13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR promoter

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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