LOCUS       sequence-139781-         813 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses SpCas9, GFP, and sgRNA targeting Ago1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 813)
  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 813)
  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_Ago1_3
FEATURES             Location/Qualifiers
     source          1..813
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             34..81
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             97..150
                     /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(151..153,154..666)
                     /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(193..214)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(454..473)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 acacggatcg acctgtctca gctgggaggc gacaaaaggc cggcggccac gaaaaaggcc
       61 ggccaggcaa aaaagaaaaa ggaattcggc agtggagagg gcagaggaag tctgctaaca
      121 tgcggtgacg tcgaggagaa tcctggccca gtgagcaagg gcgaggagct gttcaccggg
      181 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
      241 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
      301 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
      361 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
      421 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
      481 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
      541 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
      601 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
      661 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
      721 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
      781 cccaacgaga agcgcgatca catggtcctg ctg
//
