pACT2-CAS9c vector (V012296)

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LOCUS       pACT2-CAS9c.       11495 bp DNA     circular SYN 01-JAN-1980
DEFINITION  High-copy yeast plasmid for expressing S. pyogenes Cas9 under 
            control of the ADH1 promoter and terminator.
ACCESSION   .
VERSION     .
KEYWORDS    pACT2-CAS9c
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11495)
  AUTHORS   Gao Y, Zhao Y.
  TITLE     Self-processing of ribozyme-flanked RNAs into guide RNAs in vitro 
            and in vivo for CRISPR-mediated genome editing.
  JOURNAL   J Integr Plant Biol 2014;56:343-9.
  PUBMED    24373158
REFERENCE   2  (bases 1 to 11495)
  AUTHORS   Zhao Lab / Addgene #51055
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 11495)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Integr 
            Plant Biol"; date: "2014"; volume: "56"; pages: "343-9"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..11495
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        26..130
                     /label=AmpR promoter
     CDS             131..988
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1162..1750
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(1936..2076)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     promoter        2216..2614
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             2618..6721
                     /label=Cas9
                     /note="Cas9 (Csn1) endonuclease from the Streptococcus
                     pyogenes Type II CRISPR/Cas system"
     CDS             6734..6754
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T
                     antigen"
                     /note="SV40 NLS"
                     /translation="PKKKRKV"
     terminator      6890..7077
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     protein_bind    complement(7277..7310)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(7931..9022)
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
     promoter        complement(9035..9439)
                     /label=LEU2 promoter
     rep_origin      complement(9916..11258)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"