LOCUS       sequence-95568-e         624 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses Cas9 nickase and gRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 624)
  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 624)
  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_D10A-1x7
FEATURES             Location/Qualifiers
     source          1..624
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    165..186
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(201..206,207..224,225..231)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    239..255
                     /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)."
     primer_bind     244..266
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     263..279
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     263..279
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             275..580
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGL"
     primer_bind     298..318
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        300..318
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(365..384)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(366..384)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(394..411)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(394..410)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(403..425)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 aatactccac ccattgacgt caatggaaag tccctattgg cgttactatt gacgtcaatg
       61 ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgggta
      121 cctagagacc accggtggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
      181 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
      241 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca
      301 attaaccctc actaaaggga acaaaagctg gaggacctcg agggggggcc cggtacccaa
      361 ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga
      421 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
      481 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
      541 tggcgaatgg gacgcgccct gtagcggcgc cggtctctag agccatttgt ctgcagaatt
      601 ggcgcacgcg ctaaaaacgg acta
//
