LOCUS       sequence-139764-         897 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  expresses sgRNA from the U6 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 897)
  AUTHORS   Shen B, Zhang W, Zhang J, Zhou J, Wang J, Chen L, Wang L, Hodgkins 
            A, Iyer V, Huang X, Skarnes WC
  TITLE     Efficient genome modification by CRISPR-Cas9 nickase with minimal 
            off-target effects.
  JOURNAL   Nat Methods. 2014 Mar 2. doi: 10.1038/nmeth.2857.
  PUBMED    24584192
REFERENCE   2  (bases 1 to 897)
  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: "Nat 
            Methods. 2014 Mar 2. doi: 10.1038/nmeth.2857."
COMMENT     Sequence Label: pGL3-U6-sgRNA-PGK-puromycin
FEATURES             Location/Qualifiers
     source          1..897
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        86..334
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     86..106
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     257..276
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        358..433
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    485..602
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 tttctctatc gataggtacc gattagtgaa cggatctcga cggtatcgat cacgagacta
       61 gcctcgagcg gccgccccct tcaccgaggg cctatttccc atgattcctt catatttgca
      121 tatacgatac aaggctgtta gagagataat tggaattaat ttgactgtaa acacaaagat
      181 attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa
      241 attatgtttt aaaatggact atcatatgct taccgtaact tgaaagtatt tcgatttctt
      301 ggctttatat atcttgtgga aaggacgaaa caccggtgag accgagagag ggtctcagtt
      361 ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc
      421 accgagtcgg tgcttttttt aaagaattct cgacctcgag acaaatggca gtattcatcc
      481 acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca
      541 taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt
      601 ttcgggttta ttacagggac agcagagatc cactttggcc gcggctcgag ggggttgggg
      661 ttgcgccttt tccaaggcag ccctgggttt gcgcagggac gcggctgctc tgggcgtggt
      721 tccgggaaac gcagcggcgc cgaccctggg actcgcacat tcttcacgtc cgttcgcagc
      781 gtcacccgga tcttcgccgc tacccttgtg ggccccccgg cgacgcttcc tgctccgccc
      841 ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga cgtgacaaac ggaagcc
//
