LOCUS       sequence-145672-         773 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 773)
  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 773)
  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..773
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        89..337
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     89..109
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     260..279
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(466..482)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(466..482)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(519..539)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(519..537)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(558..574)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(558..574)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(571..593)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    582..598
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(606..612,613..630,631..636))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    651..672
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 atcgataagc ttgatatcgt gtacaaaaaa gcaggcttta aaggaaccaa ttcagtcgac
       61 tggatccggt accaaggtcg ggcaggaaga gggcctattt cccatgattc cttcatattt
      121 gcatatacga tacaaggctg ttagagagat aattagaatt aatttgactg taaacacaaa
      181 gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt
      241 aaaattatgt tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt
      301 cttggcttta tatatcttgt ggaaaggacg aaacaccgtt tgggggatgc tcgctccagt
      361 ttaagagcta tgctggaaac agcatagcaa gtttaaataa ggctagtccg ttatcaactt
      421 gaaaaagtgg caccgagtcg gtgctttttt tcctgcagcc cgggggatcc actagttcta
      481 gagcggccgc caccgcggtg gagctccagc ttttgttccc tttagtgagg gttaattgcg
      541 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
      601 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
      661 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
      721 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgg
//
