LOCUS       sequence-96422-e         344 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses Cas9 nuclease and gRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 344)
  AUTHORS   Sakuma T, Nishikawa A, Kume S, Chayama K, Yamamoto T
  TITLE     Multiplex genome engineering in human cells using all-in-one 
            CRISPR/Cas9 vector system.
  JOURNAL   Sci Rep. 2014 Jun 23;4:5400. doi: 10.1038/srep05400.
  PUBMED    24954249
REFERENCE   2  (bases 1 to 344)
  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: "Sci Rep."; 
            date: "2014-06-23"; volume: "4:5400. doi"; pages: " 
            10.1038/srep05400"
COMMENT     Sequence Label: pX330A-1x6
FEATURES             Location/Qualifiers
     source          1..344
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     2..19
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3..19
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     29..48
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        29..47
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(95..115)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(95..113)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(134..150)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(134..150)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(147..169)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    158..174
                     /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(182..188,189..206,207..212))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    227..248
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ttgtaaaacg acggccagtg agcgcgcgta atacgactca ctatagggcg aattgggtac
       61 cgggcccccc ctcgaggtcc tccagctttt gttcccttta gtgagggtta attgcgcgct
      121 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
      181 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
      241 tcacattaat tgcgttgcgc tcactgcccg ctttccaccg gtggtctctg aggtacccgt
      301 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacg
//
