LOCUS       sequence-163792-        1033 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  AAV backbone with a minimal H1/TO promoter driving expression of a 
            sgRNA. The gRNA can be inserted via Sapl. TracrRNA compatible with 
            SpCas9. gRNA transcription is doxycycline dependent..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1033)
  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 1033)
  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: 
            pJEP14-AAV-H1/TO-sgRNA(Empty)-CMV-TetR-P2A-eGFP-KASH-pA
FEATURES             Location/Qualifiers
     source          1..1033
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     13..37
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             164..787
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(240..259)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             806..859
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     primer_bind     complement(902..923)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 ctatataagc agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg
       61 ttttgacctc catagaagac accgggaccg atccagcctc cgcggccggg aacggtgcat
      121 tggaacggac ctgcaggaat tgatccgcgg ccgcaccggt accatgatgt ccagattaga
      181 taaaagtaaa gtgattaaca gcgcattaga gctgcttaat gaggtcggaa tcgaaggttt
      241 aacaacccgt aaactcgccc agaagctagg tgtagagcag cctacattgt attggcatgt
      301 aaaaaataag cgggctttgc tcgacgcctt agccattgag atgttagata ggcaccatac
      361 tcacttttgc cctttagaag gggaaagctg gcaagatttt ttacgtaata acgctaaaag
      421 ttttagatgt gctttactaa gtcatcgcga tggagcaaaa gtacatttag gtacacggcc
      481 tacagaaaaa cagtatgaaa ctctcgaaaa tcaattagcc tttttatgcc aacaaggttt
      541 ttcactagag aatgcattat atgcactcag cgctgtgggg cattttactt taggttgcgt
      601 attggaagat caagagcatc aagtcgctaa agaagaaagg gaaacaccta ctactgatag
      661 tatgccgcca ttattacgac aagctatcga attatttgat caccaaggtg cagagccagc
      721 cttcttattc ggccttgaat tgatcatatg cggattagaa aaacaactta aatgtgaaag
      781 tgggtccgcg gctagcggca gtggagaggg cagaggaagt ctgctaacat gcggtgacgt
      841 cgaggagaat cctggcccag tgagcaaggg cgaggagctg ttcaccgggg tggtacccat
      901 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
      961 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     1021 cgtgccctgg ccc
//
