pCas9-EQR_P4P1r vector (V016256)

LOCUS       62056_4885        8581 bp DNA     circular SYN 18-APR-2020
DEFINITION  Gateway adapted binary vector pCas9-EQR_P4P1r DNA, complete 
            sequence.
ACCESSION   LC477683
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8581)
  AUTHORS   Yamamoto A, Ishida T, Yoshimura M, Kimura Y, Sawa S.
  TITLE     Developing heritable mutations in Arabidopsis thaliana using a 
            modified CRISPR/Cas9 toolkit comprising PAM-altered Cas9 variants 
            and gRNAs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8581)
  AUTHORS   Ishida T.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-APR-2019) Contact:Takashi Ishida Kumamoto University, 
            International Research Organization for Advanced Science and 
            Technology (IROAST); Kurokami 2-39-1, Kumamoto, Kumamoto 860-8555, 
            Japan
REFERENCE   3  (bases 1 to 8581)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (18-APR-2019) Contact:Takashi Ishida Kumamoto University, 
            International Research Organization for Advanced Science and 
            Technology (IROAST); Kurokami 2-39-1, Kumamoto, Kumamoto 860-8555, 
            Japan"
FEATURES             Location/Qualifiers
     source          1..8581
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    424..519
                     /label=attL4
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      525..1496
                     /label=PcUbi promoter
                     /note="PcUbi promoter"
                     /regulatory_class="promoter"
     CDS             1511..5614
                     /label=Cas9 VQR
                     /note="Streptococcus pyogenes Cas9 endonuclease with the 
                     D1135V/R1335Q/T1337R mutations"
     regulatory      5665..6133
                     /label=rbcS-3A terminator
                     /note="rbcS-3A terminator"
                     /regulatory_class="terminator"
     protein_bind    6152..6276
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(6360..6376)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6384..6400)
                     /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(6408..6438)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6453..6474)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6762..7350)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7524..8381)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(8382..8486)
                     /label=AmpR promoter