LOCUS       sequence-162780-        1008 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of SpCas9 with 4 ERT2 fusion protein and empty gRNA 
            cassette. The activity of Cas9 can be switched on and off in human 
            cells with 4-hydroxytamoxifen (4-HT).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1008)
  AUTHORS   Liu KI, Ramli MN, Woo CW, Wang Y, Zhao T, Zhang X, Yim GR, Chong BY,
            Gowher A, Chua MZ, Jung J, Lee JH, Tan MH
  TITLE     A chemical-inducible CRISPR-Cas9 system for rapid control of genome 
            editing.
  JOURNAL   Nat Chem Biol. 2016 Sep 12. doi: 10.1038/nchembio.2179.
  PUBMED    27618190
REFERENCE   2  (bases 1 to 1008)
  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: "Nat Chem 
            Biol. 2016 Sep 12. doi: 10.1038/nchembio.2179."
COMMENT     Sequence Label: iCas
FEATURES             Location/Qualifiers
     source          1..1008
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     135..154
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        240..315
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(365..381)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(365..381)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(378..400)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    389..405
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(413..419,420..437,438..443))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    458..479
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(596..613)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(847..866)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tacgatacaa ggctgttaga gagataatta gaattaattt gactgtaaac acaaagatat
       61 tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat
      121 tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg
      181 ctttatatat cttgtggaaa ggacgaaaca ccgggtcttc gagctagcct cgaagacctg
      241 ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg
      301 gcaccgagtc ggtgcttttt tctagtatac cgtcgacctc tagctagagc ttggcgtaat
      361 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
      421 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
      481 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
      541 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
      601 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
      661 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
      721 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
      781 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
      841 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
      901 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
      961 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctcca
//
