LOCUS       sequence-107980-         721 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  empty backbone for cloning N. meningiditis protospacers. Encodes 
            constant region of N. meningiditis Cas9 synthetic gRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 721)
  AUTHORS   Hilton IB, D'Ippolito AM, Vockley CM, Thakore PI, Crawford GE, Reddy
            TE, Gersbach CA
  TITLE     Epigenome editing by a CRISPR-Cas9-based acetyltransferase activates
            genes from promoters and enhancers.
  JOURNAL   Nat Biotechnol. 2015 May;33(5):510-7. doi: 10.1038/nbt.3199. Epub 
            2015 Apr 6.
  PUBMED    25849900
REFERENCE   2  (bases 1 to 721)
  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: "10.1038/nbt.3199"; 
            journalName: "Nat Biotechnol"; date: "2015-05"; volume: "33"; issue:
            "5"; pages: "510-7"
COMMENT     Sequence Label: pZDonor-Nm-Cas9-gRNA-hU6
FEATURES             Location/Qualifiers
     source          1..721
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     157..176
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(407..427)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(407..425)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(446..462)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(446..462)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(459..481)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    470..486
                     /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(494..500,501..518,519..524))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    539..560
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(677..694)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atgattcctt catatttgca tatacgatac aaggctgtta gagagataat tggaattaat
       61 ttgactgtaa acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt
      121 gggtagtttg cagttttaaa attatgtttt aaaatggact atcatatgct taccgtaact
      181 tgaaagtatt tcgatttctt ggctttatat atcttgtgga aaggacgaaa caccgggtct
      241 tcgagaagac ctgttgtagc tccctttctc atttcggaaa cgaaatgaga accgttgcta
      301 caataaggcc gtctgaaaag atgtgccgca acgctctgcc ccttaaagct tctgctttaa
      361 ggggcttttt ttgcggccgc ccgcggtgga gctccagctt ttgttccctt tagtgagggt
      421 taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
      481 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
      541 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
      601 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
      661 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
      721 c
//
