p2U6-CAG-Cas9-p2A-puro vector (V015993)

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LOCUS       62056_1980        9989 bp DNA     circular SYN 25-JAN-2019
DEFINITION  Cloning vector p2U6-CAG-Cas9-p2A-puro, complete sequence.
ACCESSION   MK318530
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9989)
  AUTHORS   Kim J.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-DEC-2018) Biological Sciences, UCSD, 9500 Gilman Dr, 
            La Jolla, CA 92093, USA
REFERENCE   2  (bases 1 to 9989)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (18-DEC-2018) Biological Sciences, UCSD, 9500 Gilman Dr, La Jolla, 
            CA 92093, USA"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2..13
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
     regulatory      21..261
                     /label=RNA pol lll promoter for human U6 snRNA
                     /note="RNA pol lll promoter for human U6 snRNA"
                     /regulatory_class="promoter"
     misc_RNA        294..369
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        423..663
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     intron          1442..2454
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and
                     rabbit beta-globin"
     CDS             2647..6750
                     /label=Cas9
                     /note="Cas9 (Csn1) endonuclease from the Streptococcus
                     pyogenes Type II CRISPR/Cas system"
     CDS             6790..6846
                     /label=P2A
                     /note="2A peptide from porcine teschovirus-1 polyprotein"
     CDS             6847..7440
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     polyA_signal    7497..7630
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(7715..7745)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7760..7781)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(8069..8657)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8831..9688)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(9689..9793)
                     /label=AmpR promoter