Cas9m4 载体 (V012349#)

载体名称 Cas9m4 载体抗性 Ampicillin
载体长度 9553 bp 载体类型 CRISPR Plasmids
载体来源 Mali P, Aach J, Stranges PB, Esvelt KM, Moosburner M, Kosuri S, Yang 筛选标记 Neomycin/G418(Geneticin)
拷贝数 High copy number

Cas9m4 载体载体图谱

Cas9m49553 bp400800120016002000240028003200360040004400480052005600600064006800720076008000840088009200CAP binding sitelac promoterlac operatorM13 revSV40 poly(A) signalNeoR/KanRSV40 promoterf1 oriHSV TK poly(A) signalSV40 NLSCas9m4CMV promoterCMV enhancerAmpR promoterAmpRori

Cas9m4 载体载体序列

LOCUS       Cas9m4.        9553 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Plasmid for expression in mammalian cells of catalytically dead 
KEYWORDS    Cas9m4.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9553)
  AUTHORS   Mali P, Aach J, Stranges PB, Esvelt KM, Moosburner M, Kosuri S, Yang
            L, Church GM.
  TITLE     CAS9 transcriptional activators for target specificity screening and
            paired nickases for cooperative genome engineering.
  JOURNAL   Nat. Biotechnol. 2013;31:833-8.
  PUBMED    23907171
REFERENCE   2  (bases 1 to 9553)
  AUTHORS   Church Lab / Addgene #47316
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 9553)
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. 
            Biotechnol."; date: "2013"; volume: "31"; pages: "833-8"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9553
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
     polyA_signal    complement(258..391)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(570..1361)
                     /note="aminoglycoside phosphotransferase"
     promoter        complement(1428..1757)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      complement(1771..2199)
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     polyA_signal    complement(2401..2449)
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     CDS             complement(2548..2568)
                     /product="nuclear localization signal of SV40 large T
                     /note="SV40 NLS"
     CDS             complement(2581..6684)
                     /note="catalytically dead mutant of the Cas9 endonuclease
                     from the Streptococcus pyogenes Type II CRISPR/Cas system"
     promoter        complement(6799..7002)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
     enhancer        complement(7003..7382)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7648..7752
                     /label=AmpR promoter
     CDS             7753..8610
     rep_origin      8784..9372
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of