LOCUS       sequence-113816-         669 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses FokI-dCas9 and gRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 669)
  AUTHORS   Nakagawa Y, Sakuma T, Sakamoto T, Ohmuraya M, Nakagata N, Yamamoto T
  TITLE     Production of knockout mice by DNA microinjection of various 
            CRISPR/Cas9 vectors into freeze-thawed fertilized oocytes.
  JOURNAL   BMC Biotechnol. 2015 May 22;15(1):33.
  PUBMED    25997509
REFERENCE   2  (bases 1 to 669)
  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: "BMC 
            Biotechnol."; date: "2015-05-22"; volume: "15(1)"; pages: "33"
COMMENT     Sequence Label: pX330A_FokI-1x5
FEATURES             Location/Qualifiers
     source          1..669
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(60..80)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(60..78)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(99..115)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(99..115)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(112..134)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    123..139
                     /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(147..153,154..171,172..177))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    192..213
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     enhancer        264..549
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 actcactata gggcgaattg ggtaccgggc cccccctcga ggtcctccag cttttgttcc
       61 ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga
      121 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
      181 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
      241 caccggtggt ctctagaggt acccgttaca taacttacgg taaatggccc gcctggctga
      301 ccgcccaacg acccccgccc attgacgtca atagtaacgc caatagggac tttccattga
      361 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      421 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattgtgcc
      481 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      541 attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc
      601 ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg
      661 ggggggggg
//
