LOCUS       sequence-168109-         705 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Dox-inducible CRISPR interference (CRISPRi) knock in construct into 
            the AAVS1 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 705)
  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 705)
  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-NDi-CRISPRi (Gen2)
FEATURES             Location/Qualifiers
     source          1..705
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      10..19
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(35..88)
                     /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"
     misc_feature    112..137
                     /label=SA
                     /note="splice acceptor site"
     protein_bind    144..177
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 tggccgatcc catggtggcg gtaccctcga gcctagggcc gggattctcc tccacgtcac
       61 cgcatgttag aagacttcct ctgccctctc cgctgccaga tctctcgagg ccctgtggga
      121 ggaagagaag aggtcagaag cttataactt cgtataatgt atgctatacg aagttattgc
      181 cccactgtgg ggtggagggg acagataaaa gtacccagaa ccagagccac attaaccggc
      241 cctgggaata taaggtggtc ccagctcggg gacacaggat ccctggaggc agcaaacatg
      301 ctgtcctgaa gtggacatag gggcccgggt tggaggaaga agactagctg agctctcgga
      361 cccctggaag atgccatgac agggggctgg aagagctagc acagactaga gaggtaaggg
      421 gggtagggga gctgcccaaa tgaaaggagt gagaggtgac ccgaatccac aggagaacgg
      481 ggtgtccagg caaagaaagc aagaggatgg agaggtggct aaagccaggg agacggggta
      541 ctttggggtt gtccagaaaa acggtgatga tgcaggccta caagaagggg aggcgggacg
      601 caagggagac atccgtcgga gaaggccatc ctaagaaacg agagatggca caggccccag
      661 aaggagaagg aaaagggaac ccagcgagtg aagacggcat ggggt
//
