LOCUS       sequence-107981-         704 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 704)
  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 704)
  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..704
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(165..413)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(223..242)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(429..448)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(430..448)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(458..475)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(458..474)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(467..489)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 gctggagctc caccgcgggc ggccgcaaaa aaagcccctt aaagcagaag ctttaagggg
       61 cagagcgttg cggcacatct tttcagacgg ccttattgta gcaacggttc tcatttcgtt
      121 tccgaaatga gaaagggagc tacaacaggt cttctcgaag acccggtgtt tcgtcctttc
      181 cacaagatat ataaagccaa gaaatcgaaa tactttcaag ttacggtaag catatgatag
      241 tccattttaa aacataattt taaaactgca aactacccaa gaaattatta ctttctacgt
      301 cacgtatttt gtactaatat ctttgtgttt acagtcaaat taattccaat tatctctcta
      361 acagccttgt atcgtatatg caaatatgaa ggaatcatgg gaaataggca agcttggtac
      421 ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc
      481 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
      541 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
      601 tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
      661 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgct
//
