LOCUS       sequence-150782-         860 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  All-in-one CRISPR/Cas9 vector with high-fidelity eSpCas9 expression 
            in human pluripotent stem cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 860)
  TITLE     pCAG-SpCas9-U6-gRNA
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 860)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pCAG-eCas9-GFP-U6-gRNA
FEATURES             Location/Qualifiers
     source          1..860
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     226..247
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(290..343)
                     /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             complement(359..406)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
ORIGIN
        1 tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt
       61 ggtcggggta gcggctgaag cactgcacgc cgtaggtcag ggtggtcacg agggtgggcc
      121 agggcacggg cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg
      181 catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct
      241 cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcact gggccaggat
      301 tctcctcgac gtcaccgcat gttagcagac ttcctctgcc ctctccactg ccgaattcct
      361 ttttcttttt tgcctggccg gcctttttcg tggccgccgg ccttttgtcg cctcccagct
      421 gagacaggtc gatccgtgtc tcgtacaggc cggtgatgct ctggtggatc agggtggcgt
      481 ccagcacctc tttggtgctg gtgtacctct tccggtcgat ggtggtgtca aagtacttga
      541 aggcggcagg ggctcccaga ttggtcaggg taaacaggtg gatgatattc tcggcctgct
      601 ctctgatggg cttatcccgg tgcttgttgt aggcggacag cactttgtcc agattagcgt
      661 cggccaggat cactctcttg gagaactcgc tgatctgctc gatgatctcg tccaggtagt
      721 gcttgtgctg ttccacaaac agctgtttct gctcattatc ctcgggggag cccttcagct
      781 tctcatagtg gctggccagg tacaggaagt tcacatattt ggagggcagg gccagttcgt
      841 ttcccttctg cagttcgccg
//
