LOCUS       sequence-121540-         534 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  U6 driven sgRNA cloning vector where guide sequences are inserted 
            between BfuAI sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 534)
  AUTHORS   Kearns NA, Genga RM, Enuameh MS, Garber M, Wolfe SA, Maehr R
  TITLE     Cas9 effector-mediated regulation of transcription and 
            differentiation in human pluripotent stem cells.
  JOURNAL   Development. 2014 Jan;141(1):219-23. doi: 10.1242/dev.103341.
  PUBMED    24346702
REFERENCE   2  (bases 1 to 534)
  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: "10.1242/dev"; 
            journalName: "Development"; date: "2014-01"; volume: "141"; issue: 
            "1"; pages: "219-23"
COMMENT     Sequence Label: pLKO.1-puro U6 sgRNA BfuAI stuffer
FEATURES             Location/Qualifiers
     source          1..534
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        59..307
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     59..79
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     230..249
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        341..416
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 gaacggatct cgacggtatc gatcacgaga ctagcctcga gcggccgccc ccttcaccga
       61 gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg ttagagagat
      121 aattggaatt aatttgactg taaacacaaa gatattagta caaaatacgt gacgtagaaa
      181 gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg actatcatat
      241 gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt ggaaaggacg
      301 aaacaccggt gtgcaggtga gtgatcaaga acctgctgac gttttagagc tagaaatagc
      361 aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt
      421 tttgaattct cgacctcgag acaaatggca gtattcatcc acaattttaa aagaaaaggg
      481 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agac
//
