LOCUS       sequence-139763-         779 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses SpCas9, GFP, and sgRNA targeting Ago2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 779)
  AUTHORS   Ngondo RP, Cirera-Salinas D, Yu J, Wischnewski H, Bodak M, 
            Vandormael-Pournin S, Geiselmann A, Wettstein R, Luitz J, 
            Cohen-Tannoudji M, Ciaudo C
  TITLE     Argonaute 2 Is Required for Extra-embryonic Endoderm Differentiation
            of Mouse Embryonic Stem Cells.
  JOURNAL   Stem Cell Reports. 2018 Jan 24. pii: S2213-6711(17)30571-4. doi: 
            10.1016/j.stemcr.2017.12.023.
  PUBMED    29396181
REFERENCE   2  (bases 1 to 779)
  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: "Stem Cell 
            Reports. 2018 Jan 24. pii: S2213-6711(17)30571-4. doi: 
            10.1016/j.stemcr.2017.12.023."
COMMENT     Sequence Label: pX458-sgRNA_Ago2_4
FEATURES             Location/Qualifiers
     source          1..779
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             38..85
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             101..154
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(155..157,158..670)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIE"
     primer_bind     complement(197..218)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(458..477)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 cgagacacgg atcgacctgt ctcagctggg aggcgacaaa aggccggcgg ccacgaaaaa
       61 ggccggccag gcaaaaaaga aaaaggaatt cggcagtgga gagggcagag gaagtctgct
      121 aacatgcggt gacgtcgagg agaatcctgg cccagtgagc aagggcgagg agctgttcac
      181 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
      241 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
      301 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
      361 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
      421 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
      481 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
      541 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
      601 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
      661 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
      721 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagca
//
