LOCUS       sequence-64960-e         877 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Dual expression plasmid of human codon-optimized SpCas9 and a gRNA 
            to the human Emx1 locus, can be used to test SpCas9 cleavage in cell
            lines of choice..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 877)
  AUTHORS   Cong L, Ran FA, Cox D, Lin S, Barretto R, Habib N, Hsu PD, Wu X, 
            Jiang W, Marraffini LA, Zhang F
  TITLE     Multiplex Genome Engineering Using CRISPR/Cas Systems.
  JOURNAL   Science. 2013 Jan 3.
  PUBMED    23287718
REFERENCE   2  (bases 1 to 877)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science. 
            2013 Jan 3."
COMMENT     Sequence Label: 
            pX261-U6-DR-hEmx1-DR-Cbh-NLS-hSpCas9-NLS-H1-shorttracr-PGK-puro
FEATURES             Location/Qualifiers
     source          1..877
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        123..371
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     123..143
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     294..313
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     repeat_region   374..409
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     repeat_region   440..475
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     enhancer        526..811
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
       61 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
      121 gtgagggcct atttcccatg attccttcat atttgcatat acgatacaag gctgttagag
      181 agataattgg aattaatttg actgtaaaca caaagatatt agtacaaaat acgtgacgta
      241 gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa atggactatc
      301 atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc ttgtggaaag
      361 gacgaaacac cgggttttag agctatgctg ttttgaatgg tcccaaaacg gaagggcctg
      421 agtccgagca gaagaagaag ttttagagct atgctgtttt gaatggtccc aaaacttttt
      481 ctagcgcgtg cgccaattct gcagacaaat ggctctagag gtacccgtta cataacttac
      541 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caatagtaac
      601 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
      661 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
      721 atggcccgcc tggcattgtg cccagtacat gaccttatgg gactttccta cttggcagta
      781 catctacgta ttagtcatcg ctattaccat ggtcgaggtg agccccacgt tctgcttcac
      841 tctccccatc tcccccccct ccccaccccc aattttg
//
