LOCUS       sequence-113751-        1153 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  5' and 3' EGFP fragments that shares 482bp were placed under 
            ubiquitous CAG promoter. Used for validation of gRNA sequences by 
            DSB mediated EGFP reconstitution. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1153)
  AUTHORS   Mashiko D, Fujihara Y, Satouh Y, Miyata H, Isotani A, Ikawa M
  TITLE     Generation of mutant mice by pronuclear injection of circular 
            plasmid expressing Cas9 and single guided RNA.
  JOURNAL   Sci Rep. 2013 Nov 27;3:3355. doi: 10.1038/srep03355.
  PUBMED    24284873
REFERENCE   2  (bases 1 to 1153)
  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: "2013-11-27"; volume: "3:3355. doi"; pages: " 
            10.1038/srep03355"
COMMENT     Sequence Label: pCAG-EGxxFP
FEATURES             Location/Qualifiers
     source          1..1153
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     49..68
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      96..105
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(102..104,105..107,108..620)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIE"
     primer_bind     complement(147..168)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg
       61 ttattgtgct gtctcatcat tttggcaaat ctagagccgc catggtgagc aagggcgagg
      121 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
      181 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
      241 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
      301 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
      361 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
      421 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
      481 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
      541 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
      601 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
      661 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctgaggatcc gctagcctgc
      721 aggtcgacga attcgatatc ggcaagctga ccctgaagtt catctgcacc accggcaagc
      781 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
      841 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
      901 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
      961 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     1021 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     1081 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     1141 acggcagcgt gca
//
