LOCUS       sequence-151775-         708 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 708)
  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 708)
  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..708
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      13..22
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             22..87
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             94..114
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 ggttggaccg gtgccaccat ggactataag gaccacgacg gagactacaa ggatcatgat
       61 attgattaca aagacgatga cgataagatg gccccaaaga agaagcggaa ggtcggtatc
      121 cacggagtcc cagcagccga caagaagtac agcatcggcc tggacatcgg caccaactct
      181 gtgggctggg ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt caaggtgctg
      241 ggcaacaccg accggcacag catcaagaag aacctgatcg gagccctgct gttcgacagc
      301 ggcgaaacag ccgaggccac ccggctgaag agaaccgcca gaagaagata caccagacgg
      361 aagaaccgga tctgctatct gcaagagatc ttcagcaacg agatggccaa ggtggacgac
      421 agcttcttcc acagactgga agagtccttc ctggtggaag aggataagaa gcacgagcgg
      481 caccccatct tcggcaacat cgtggacgag gtggcctacc acgagaagta ccccaccatc
      541 taccacctga gaaagaaact ggtggacagc accgacaagg ccgacctgcg gctgatctat
      601 ctggccctgg cccacatgat caagttccgg ggccacttcc tgatcgaggg cgacctgaac
      661 cccgacaaca gcgacgtgga caagctgttc atccagctgg tgcagacc
//
