LOCUS       sequence-168100-         932 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Dox-inducible CRISPR nuclease (CRISPRn) knock in construct into the 
            AAVS1 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 932)
  AUTHORS   Mandegar MA, Huebsch N, Frolov EB, Shin E, Truong A, Olvera MP, Chan
            AH, Miyaoka Y, Holmes K, Spencer CI, Judge LM, Gordon DE, Eskildsen 
            TV, Villalta JE, Horlbeck MA, Gilbert LA, Krogan NJ, Sheikh SP, 
            Weissman JS, Qi LS, So PL, Conklin BR
  TITLE     CRISPR Interference Efficiently Induces Specific and Reversible Gene
            Silencing in Human iPSCs.
  JOURNAL   Cell Stem Cell. 2016 Apr 7;18(4):541-53. doi: 
            10.1016/j.stem.2016.01.022. Epub 2016 Mar 10.
  PUBMED    26971820
REFERENCE   2  (bases 1 to 932)
  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: 
            "10.1016/j.stem.2016.01.022"; journalName: "Cell Stem Cell"; date: 
            "2016-04-7- 7"; volume: "18"; issue: "4"; pages: "541-53"
COMMENT     Sequence Label: pAAVS1-PDi-CRISPRn
FEATURES             Location/Qualifiers
     source          1..932
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(311..331)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(338..403)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      403..412
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(496..520)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    565..583
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    601..619
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    637..655
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    673..691
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    709..727
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    745..763
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    781..799
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 agctgtcgtc caccttggcc atctcgttgc tgaagatctc ttgcagatag cagatccggt
       61 tcttccgtct ggtgtatctt cttctggcgg ttctcttcag ccgggtggcc tcggctgttt
      121 cgccgctgtc gaacagcagg gctccgatca ggttcttctt gatgctgtgc cggtcggtgt
      181 tgcccagcac cttgaatttc ttgctgggca ccttgtactc gtcggtgatc acggcccagc
      241 ccacagagtt ggtgccgatg tccaggccga tgctgtactt cttgtcggct gctgggactc
      301 cgtggatacc gaccttccgc ttcttctttg gggccatctt atcgtcatcg tctttgtaat
      361 caatatcatg atccttgtag tctccgtcgt ggtccttata gtccatggtg gcgttaatta
      421 accttaagtt tacgagggta ggaagtggta cggaaagttg gtataagaca aaagtgttgt
      481 ggaattgctc caggcgatct gacggttcac taaacgagct ctgcttttat aggcgcccac
      541 cgtacacgcc taaagcttat acgttctcta tcactgatag ggagtaaact ggatatacgt
      601 tctctatcac tgatagggag taaactgtag atacgttctc tatcactgat agggagtaaa
      661 ctggtcatac gttctctatc actgataggg agtaaactcc ttatacgttc tctatcactg
      721 atagggagta aagtctgcat acgttctcta tcactgatag ggagtaaact cttcatacgt
      781 tctctatcac tgatagggag taaaccactg tggggtggag atatcagctt tactagggac
      841 aggattggtg acagaaaagc cccatcctta ggcctcctcc ttcctagtct cctgatattg
      901 ggtctaaccc ccacctcctg ttaggcagat tc
//
