LOCUS       sequence-153350-         550 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 550)
  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 550)
  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-1x2
FEATURES             Location/Qualifiers
     source          1..550
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    170..191
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(206..211,212..229,230..236)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    244..260
                     /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     249..271
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     268..284
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     268..284
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             280..549
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNG"
     primer_bind     303..323
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        305..323
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(370..389)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(371..389)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(399..416)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(399..415)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(408..430)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 aatactccac ccattgacgt caatggaaag tccctattgg cgttactatt gacgtcaatg
       61 ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgggta
      121 cctctagaag agaccaccgg tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
      181 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
      241 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
      301 gcgcaattaa ccctcactaa agggaacaaa agctggagga cctcgagggg gggcccggta
      361 cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt
      421 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
      481 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
      541 ctgaatggcg
//
