LOCUS       sequence-145673-         799 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Encodes an sgRNA for spCas9 driven by hU6 promoter with a modified 
            scaffold (Chen et al. Cell 2013) and a spacer targeting the 3'UTR of
            ACTB mRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 799)
  AUTHORS   Nelles DA, Fang MY, O'Connell MR, Xu JL, Markmiller SJ, Doudna JA, 
            Yeo GW
  TITLE     Programmable RNA Tracking in Live Cells with CRISPR/Cas9.
  JOURNAL   Cell. 2016 Mar 16. pii: S0092-8674(16)30204-5. doi: 
            10.1016/j.cell.2016.02.054.
  PUBMED    26997482
REFERENCE   2  (bases 1 to 799)
  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: "Cell. 2016 
            Mar 16. pii: S0092-8674(16)30204-5. doi: 
            10.1016/j.cell.2016.02.054."
COMMENT     Sequence Label: pBluescriptSKII+ U6-sgRNA(F+E) ACTB
FEATURES             Location/Qualifiers
     source          1..799
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(140..388)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(198..217)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(368..388)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(473..489)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(474..490)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(514..533)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(515..533)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(543..560)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(543..559)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(552..574)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 actagtggat cccccgggct gcaggaaaaa aagcaccgac tcggtgccac tttttcaagt
       61 tgataacgga ctagccttat ttaaacttgc tatgctgttt ccagcatagc tcttaaactg
      121 gagcgagcat cccccaaacg gtgtttcgtc ctttccacaa gatatataaa gccaagaaat
      181 cgaaatactt tcaagttacg gtaagcatat gatagtccat tttaaaacat aattttaaaa
      241 ctgcaaacta cccaagaaat tattactttc tacgtcacgt attttgtact aatatctttg
      301 tgtttacagt caaattaatt ctaattatct ctctaacagc cttgtatcgt atatgcaaat
      361 atgaaggaat catgggaaat aggccctctt cctgcccgac cttggtaccg gatccagtcg
      421 actgaattgg ttcctttaaa gcctgctttt ttgtacacga tatcaagctt atcgataccg
      481 tcgacctcga gggggggccc ggtacccaat tcgccctata gtgagtcgta ttacgcgcgc
      541 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
      601 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
      661 cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca
      721 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
      781 gcgcccgctc ctttcgctt
//
