LOCUS       sequence-112317-         620 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Vector for tandem expression of two sgRNAs from two independent U6 
            promoters. Cas9 is expressed by Cbh promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 620)
  AUTHORS   Maddalo D, Manchado E, Concepcion CP, Bonetti C, Vidigal JA, Han YC,
            Ogrodowski P, Crippa A, Rekhtman N, de Stanchina E, Lowe SW, Ventura
            A
  TITLE     In vivo engineering of oncogenic chromosomal rearrangements with the
            CRISPR/Cas9 system.
  JOURNAL   Nature. 2014 Dec 18;516(7531):423-7. doi: 10.1038/nature13902. Epub 
            2014 Oct 22.
  PUBMED    25337876
REFERENCE   2  (bases 1 to 620)
  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"; doi: 
            "10.1038/nature13902"; journalName: "Nature"; date: "2014-12-18- 
            18"; volume: "516"; issue: "7531"; pages: "423-7"
COMMENT     Sequence Label: pX333
FEATURES             Location/Qualifiers
     source          1..620
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_RNA        complement(146..221)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(248..496)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(306..325)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(476..496)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgggta
       61 ccccatttgt ctgcagaatt ggcgcacgcg ctaaaaacgg actagcctta ttttaacttg
      121 ctatttctag ctctaaaaca aaaaagcacc gactcggtgc cactttttca agttgataac
      181 ggactagcct tattttaact tgctatttct agctctaaaa ccgagacctt aaggcgaata
      241 ggtctccggt gtttcgtcct ttccacaaga tatataaagc caagaaatcg aaatactttc
      301 aagttacggt aagcatatga tagtccattt taaaacataa ttttaaaact gcaaactacc
      361 caagaaatta ttactttcta cgtcacgtat tttgtactaa tatctttgtg tttacagtca
      421 aattaattcc aattatctct ctaacagcct tgtatcgtat atgcaaatat gaaggaatca
      481 tgggaaatag gccctctcta gagccatttg tctgcagaat tggcgcacgc gctaaaaacg
      541 gactagcctt attttaactt gctatttcta gctctaaaac aaaaaagcac cgactcggtg
      601 ccactttttc aagttgataa
//
