LOCUS       sequence-139761-         818 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 818)
  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 818)
  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_3
FEATURES             Location/Qualifiers
     source          1..818
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             28..75
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             91..144
                     /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(145..147,148..660)
                     /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(187..208)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(448..467)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     795..816
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 atcgacctgt ctcagctggg aggcgacaaa aggccggcgg ccacgaaaaa ggccggccag
       61 gcaaaaaaga aaaaggaatt cggcagtgga gagggcagag gaagtctgct aacatgcggt
      121 gacgtcgagg agaatcctgg cccagtgagc aagggcgagg agctgttcac cggggtggtg
      181 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
      241 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
      301 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
      361 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
      421 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
      481 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
      541 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
      601 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
      661 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
      721 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
      781 gagaagcgcg atcacatggt cctgctggag ttcgtgac
//
