LOCUS       sequence-152735-         953 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 BbsI sites that allow insertion
            of a spacer sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 953)
  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 953)
  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) empty
FEATURES             Location/Qualifiers
     source          1..953
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     216..233
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    350..371
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(386..391,392..409,410..416)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    424..440
                     /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)."
     primer_bind     429..451
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     448..464
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     448..464
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     483..503
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        485..503
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     540..556
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     540..556
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(682..930)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(740..759)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(910..930)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
       61 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
      121 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
      181 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
      241 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
      301 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
      361 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
      421 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
      481 gcgcaattaa ccctcactaa agggaacaaa agctggagct ccaccgcggt ggcggccgct
      541 ctagaactag tggatccccc gggctgcagg aaaaaaagca ccgactcggt gccacttttt
      601 caagttgata acggactagc cttatttaaa cttgctatgc tgtttccagc atagctctta
      661 aacaggtctt ctcgaagacc cggtgtttcg tcctttccac aagatatata aagccaagaa
      721 atcgaaatac tttcaagtta cggtaagcat atgatagtcc attttaaaac ataattttaa
      781 aactgcaaac tacccaagaa attattactt tctacgtcac gtattttgta ctaatatctt
      841 tgtgtttaca gtcaaattaa ttctaattat ctctctaaca gccttgtatc gtatatgcaa
      901 atatgaagga atcatgggaa ataggccctc ttcctgcccg accttggtac cgg
//
