LOCUS       sequence-74593-e         862 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of a catalytically inactive, human codon-optimized 
            Cas9-BFP fusion under the control of Murine Stem Cell retroVirus LTR
            promoter for mammalian gene knockdown.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 862)
  AUTHORS   Qi LS, Larson MH, Gilbert LA, Doudna JA, Weissman JS, Arkin AP, Lim 
            WA
  TITLE     Repurposing CRISPR as an RNA-Guided Platform for Sequence-Specific 
            Control of Gene Expression.
  JOURNAL   Cell. 2013 Feb 28;152(5):1173-83. doi: 10.1016/j.cell.2013.02.022.
  PUBMED    23452860
REFERENCE   2  (bases 1 to 862)
  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"; doi: 
            "10.1016/j.cell.2013.02"; journalName: "Cell"; date: "2013-02-28- 
            28"; volume: "152"; issue: "5"; pages: "1173-83"
COMMENT     Sequence Label: pdCas9::BFP-humanized
FEATURES             Location/Qualifiers
     source          1..862
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(67..87)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(97..792)
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     CDS             complement(802..822)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 aagcgcatgc tccagactgc cttgggaaaa gcgcctcccc tacccggtag aattctagga
       61 actacctacc ttacgcttct tctttggagc tccctcatta agcttgtgcc ccagtttgct
      121 agggaggtcg cagtatctgg ccactgccac ctcgtgctgc tcgacgtagg tctcgttgtt
      181 ggcctccttg attctttcca gtctgtagtc cacatagtag acgccaggca tcttgaggtt
      241 cttagcgggt ttcttggatc tatatgtggt cttgatgttt gcgatcagat ggctcccgcc
      301 cacgagcttc agggccatgt cgtttctgcc ttccaggccg ccgtcagcgg ggtacagcgt
      361 ctcggtgaag gcctcccagc cgagtgtttt cttctgcatc acagggccgt tggatgtgaa
      421 gttcacccct ctgatcttga cgttgtagat gaggcagccg tcctggaggc tggtgtcctg
      481 ggtagcggtc agcacgcccc cgtcttcgta tgtggtgact ctctcccatg tgaagccctc
      541 agggaaggac tgcttgaaga agtcggggat gccctgggtg tggttgatga aggtcttgct
      601 gccgtagagg aagctagtag ccaggatgtc gaaggcgaag gggagagggc cgccctcgac
      661 caccttgatt ctcatggtct gggtgccctc gtagggcttg ccttcgccct cggatgtgca
      721 cttgaagtga tggttgtcca cggtgccctc catgtacagc ttcatgtgca tgttctcctt
      781 aatcagctcg ctggaactac ctaccttgcg ctttttcttg ggagctccct catcccctcc
      841 gagctgtgag aggtcaatcc ga
//
