LOCUS       sequence-137356-         646 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 646)
  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 646)
  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..646
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        169..417
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     340..359
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        436..511
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
       61 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgcgcc
      121 attctcgagc atgcgccatt ctcgagcatg ctcgagaatg gcgcatgtga gggcctattt
      181 cccatgattc cttcatattt gcatatacga tacaaggctg ttagagagat aattggaatt
      241 aatttgactg taaacacaaa gatattagta caaaatacgt gacgtagaaa gtaataattt
      301 cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg actatcatat gcttaccgta
      361 acttgaaagt atttcgattt cttggcttta tatatcttgt ggaaaggacg aaacaccggg
      421 tcttcgagaa gacctgtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta
      481 tcaacttgaa aaagtggcac cgagtcggtg cttttttgtt ttagagctag aaatagcaag
      541 ttaaaataag gctagtccgt ttttagcgcg tgcgccaatt ctgcagacaa atggctctag
      601 agagggccta tttcccatga ttccttcata tttgcatata cgatac
//
