LOCUS       sequence-100665-        1010 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses a guide RNA (gRNA) to target human IRF-1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1010)
  AUTHORS   Fujita T, Fujii H
  TITLE     Efficient isolation of specific genomic regions and identification 
            of associated proteins by engineered DNA-binding molecule-mediated 
            chromatin immunoprecipitation (enChIP) using CRISPR.
  JOURNAL   Biochem Biophys Res Commun. 2013 Sep 13;439(1):132-6. doi: 
            10.1016/j.bbrc.2013.08.013. Epub 2013 Aug 11.
  PUBMED    23942116
REFERENCE   2  (bases 1 to 1010)
  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.bbrc.2013.08.013"; journalName: "Biochem Biophys Res 
            Commun"; date: "2013-09-13- 13"; volume: "439"; issue: "1"; pages: 
            "132-6"
COMMENT     Sequence Label: gRNA-hIRF-1 #12
FEATURES             Location/Qualifiers
     source          1..1010
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        147..395
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     147..167
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     318..337
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        416..491
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(503..527)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(612..630)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(613..630)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(648..664)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(648..664)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(661..683)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    672..688
                     /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(696..702,703..720,721..726))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    741..762
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(879..896)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ctatagggcg aattgggccc tctagatgca tgctcgagcg gccgccagtg tgatggatat
       61 ctgcagaatt cgccctttgt acaaaaaagc aggctttaaa ggaaccaatt cagtcgactg
      121 gatccggtac caaggtcggg caggaagagg gcctatttcc catgattcct tcatatttgc
      181 atatacgata caaggctgtt agagagataa ttagaattaa tttgactgta aacacaaaga
      241 tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa
      301 aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct
      361 tggctttata tatcttgtgg aaaggacgaa acaccgcggg ggcgctgggc tgtccgtttt
      421 agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac
      481 cgagtcggtg ctttttttct agacccagct ttcttgtaca aagttggcat taaagggcga
      541 attccagcac actggcggcc gttactagtg gatccgagct cggtaccaag cttgatgcat
      601 agcttgagta ttctatagtg tcacctaaat agcttggcgt aatcatggtc atagctgttt
      661 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
      721 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
      781 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
      841 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
      901 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
      961 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
//
