LOCUS       sequence-163562-         575 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  AAV backbone with a full length U6/TO promoter driving expression of
            a sgRNA that targets exon 3 of the mouse Tet2 locus. TracrRNA 
            compatible with SpCas9. gRNA transcription is doxycycline 
            dependent..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 575)
  AUTHORS   de Solis CA, Ho A, Holehonnur R, Ploski JE
  TITLE     The Development of a Viral Mediated CRISPR/Cas9 System with 
            Doxycycline Dependent gRNA Expression for Inducible In vitro and In 
            vivo Genome Editing.
  JOURNAL   Front Mol Neurosci. 2016 Aug 18;9:70. doi: 10.3389/fnmol.2016.00070.
            eCollection 2016.
  PUBMED    27587996
REFERENCE   2  (bases 1 to 575)
  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: "Front Mol 
            Neurosci."; date: "2016-08-18"; volume: "9:70. doi"; pages: " 
            10.3389/fnmol.2016.00070. eCollection 2016"
COMMENT     Sequence Label: 
            pJEP11-AAV-U6/TO-sgRNA(Tet2)-CMV-TetR-P2A-eGFP-KASH-pA
FEATURES             Location/Qualifiers
     source          1..575
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_RNA        complement(190..265)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(290..384)
                     /label=H1-2O2 promoter
                     /note="doxycycline-inducible variant of the human H1 RNA 
                     promoter (Henriksen et al., 2007)"
     protein_bind    290..308
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    318..336
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(354..373)
                     /label=H1
                     /note="Human H1 promoter, forward primer"
ORIGIN
        1 gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt
       61 ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt
      121 cagccaggcg ggccatttac cgtaagttat gtaacggact ctagaactag tccatggacc
      181 ggtaaaaaag caccgactcg gtgccacttt ttcaagttga taacggacta gccttatttt
      241 aacttgctat ttctagctct aaaacggata tctgttggca ctttcggtgt ctctatcact
      301 gatagggaac ttataagtct ctatcactga tagggatttc acgtttatgg tgatttccca
      361 gaacacatag cgacatgcaa atattgcagg gcgccactcc cctgtccctc acagccatct
      421 tcctgccagg gcgcacgcgc gctgggtgtt cccgcctagt gacactgggc ccgcgattcc
      481 ttggagcggg ttgatgacgt cagcgttcga attcacgcgt gcggccgcag gaacccctag
      541 tgatggagtt ggccactccc tctctgcgcg ctcgc
//
