LOCUS       sequence-139779-         836 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 836)
  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 836)
  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_2
FEATURES             Location/Qualifiers
     source          1..836
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             37..84
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             100..153
                     /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(154..156,157..669)
                     /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(196..217)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(457..476)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     804..825
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gagacacgga tcgacctgtc tcagctggga ggcgacaaaa ggccggcggc cacgaaaaag
       61 gccggccagg caaaaaagaa aaaggaattc ggcagtggag agggcagagg aagtctgcta
      121 acatgcggtg acgtcgagga gaatcctggc ccagtgagca agggcgagga gctgttcacc
      181 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
      241 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
      301 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
      361 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
      421 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
      481 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
      541 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
      601 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
      661 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
      721 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
      781 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgg
//
