LOCUS       sequence-168117-         915 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Dox-inducible CRISPR interference (CRISPRi) knock in construct into 
            the AAVS1 locus with mCherry marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 915)
  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 915)
  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 (Gen1)
FEATURES             Location/Qualifiers
     source          1..915
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(310..495)
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEP"
     regulatory      522..531
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(615..639)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    684..702
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    720..738
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    756..774
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    792..810
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    828..846
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    864..882
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 gctgtcgtcc accttggcca tctcgttgct gaagatctct tgcagatagc agatccggtt
       61 cttccgtctg gtgtatcttc ttctggcggt tctcttcagc cgggtggcct cggctgtttc
      121 tccgctgtcg aacagcaggg cgccgatcag gttcttcttg atgctgtgcc ggtcggtgtt
      181 gcccagcacc ttgaatttct tgctgggcac cttgtactcg tcggtgatca cggcccagcc
      241 cacagagttg gtgccgatgg ccaggccgat gctgtacttc ttgtccatcg accctccgcc
      301 ggaaccgccg ggctcttctc ccttctccaa ccggaggatc acatctggct tagtaagctg
      361 ataacccaag gaaaccaggt tcttatagtt ctccagcatc acatttctgt acacgatctg
      421 ctgagcagtg tccagcagct tccactcctc cctggtgaag tccacaaata catccttgaa
      481 ggtcaccagt gtccgggacc aggcagttag tgacttagca tccatggtgg cgttaattaa
      541 ccttaagttt acgagggtag gaagtggtac ggaaagttgg tataagacaa aagtgttgtg
      601 gaattgctcc aggcgatctg acggttcact aaacgagctc tgcttttata ggcgcccacc
      661 gtacacgcct aaagcttata cgttctctat cactgatagg gagtaaactg gatatacgtt
      721 ctctatcact gatagggagt aaactgtaga tacgttctct atcactgata gggagtaaac
      781 tggtcatacg ttctctatca ctgataggga gtaaactcct tatacgttct ctatcactga
      841 tagggagtaa agtctgcata cgttctctat cactgatagg gagtaaactc ttcatacgtt
      901 ctctatcact gatag
//
