LOCUS       sequence-113845-         644 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses dCas9 and gRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 644)
  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 644)
  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_dCas9-1x5
FEATURES             Location/Qualifiers
     source          1..644
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(59..79)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(59..77)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(98..114)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(98..114)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(111..133)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    122..138
                     /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(146..152,153..170,171..176))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    191..212
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     enhancer        263..548
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 actactatag ggcgaattgg gtaccgggcc ccccctcgag gtcctccagc ttttgttccc
       61 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
      121 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
      181 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
      241 accggtggtc tctagaggta cccgttacat aacttacggt aaatggcccg cctggctgac
      301 cgcccaacga cccccgccca ttgacgtcaa tagtaacgcc aatagggact ttccattgac
      361 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      421 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattgtgccc
      481 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      541 ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc
      601 cacccccaat tttgtattta tttatttttt aattattttg tgca
//
