LOCUS       sequence-111606-         648 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 648)
  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 648)
  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_dCas9-1x4
FEATURES             Location/Qualifiers
     source          1..648
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(62..82)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(62..80)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(101..117)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(101..117)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(114..136)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    125..141
                     /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(149..155,156..173,174..179))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    194..215
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     enhancer        267..552
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 cgactcacta tagggcgaat tgggtaccgg gccccccctc gaggtcctcc agcttttgtt
       61 ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt
      121 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
      181 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
      241 tccaccggtg gtctcttaga ggtacccgtt acataactta cggtaaatgg cccgcctggc
      301 tgaccgccca acgacccccg cccattgacg tcaatagtaa cgccaatagg gactttccat
      361 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      421 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattgt
      481 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      541 gctattacca tggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc
      601 tccccacccc caattttgta tttatttatt ttttaattat tttgtgca
//
