LOCUS       sequence-72757-e         997 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Co-expression of human codon-optimized Cas9 (D10A) mutant nickase 
            and GFP, plasmid optimized for expression in human pluripotent stem 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 997)
  AUTHORS   Ding Q, Regan SN, Xia Y, Oostrom LA, Cowan CA, Musunuru K
  TITLE     Enhanced efficiency of human pluripotent stem cell genome editing 
            through replacing TALENs with CRISPRs.
  JOURNAL   Cell Stem Cell. 2013 Apr 4;12(4):393-4. doi: 
            10.1016/j.stem.2013.03.006.
  PUBMED    23561441
REFERENCE   2  (bases 1 to 997)
  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"; doi: 
            "10.1016/j.stem.2013.03"; journalName: "Cell Stem Cell"; date: 
            "2013-04-4- 4"; volume: "12"; issue: "4"; pages: "393-4"
COMMENT     Sequence Label: pCas9D10A_GFP
FEATURES             Location/Qualifiers
     source          1..997
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(34..87)
                     /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(103..123)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 cccggtgaac agctcctcgc ccttgctcac cattgggccg ggattctcct ccacgtcacc
       61 gcatgttaga agacttcctc tgccctctcc gccgccggac ctcaccttcc tcttcttctt
      121 ggggtcagcc ctgctgtctc caccgagctg agagaggtcg attcttgttt catagagccc
      181 cgtaattgac tgatgaatca gtgtggcgtc caggacctcc tttgtagagg tgtaccgctt
      241 tctgtctatg gtggtgtcga agtacttgaa ggctgcaggc gcgcccaagt tggtcagagt
      301 aaacaagtgg ataatgtttt ctgcctgctc cctgatgggc ttatccctgt gcttattgta
      361 agcagaaagc accttatcga ggttagcgtc ggcgaggatc actcttttgg agaattcgct
      421 tatttgctcg atgatctcat caaggtagtg tttgtgttgt tccacgaaca gctgcttctg
      481 ctcattatct tcgggagacc ctttgagctt ttcatagtgg ctggccagat acaagaaatt
      541 aacgtattta gagggcagtg ccagctcgtt acctttctgc agctcgcccg cactagcgag
      601 cattcgtttc cggccgtttt caagctcaaa gagagagtac ttgggaagct taatgatgag
      661 gtcttttttg acctctttat atcctttcgc ctcgagaaag tcgatggggt ttttttcgaa
      721 gcttgatcgc tccatgattg tgatgcccag cagttccttg acgcttttga gttttttaga
      781 cttccctttc tccactttgg ccacaaccag tacactgtaa gcgactgtag gagaatcgaa
      841 tccgccgtat ttcttggggt cccaatcttt tttgcgtgcg atcagcttgt cgctgttcct
      901 tttcgggagg atactttcct tggagaagcc tccggtctgt acttcggtct ttttaacgat
      961 gttcacctgc ggcatggaca ggaccttccg gactgtc
//
