LOCUS       sequence-112394-         974 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Co-expression of human codon-optimized Staphylococcus aureus Cas9 
            nuclease and GFP, plasmid optimized for expression in human 
            pluripotent stem cells and other mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 974)
  TITLE     Staphylococcus aureus Cas9
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 974)
  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            "Unpublished"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: pSaCas9_GFP
FEATURES             Location/Qualifiers
     source          1..974
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(join(63..776,777..779,780..782))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(66..776,777..779,780..782))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(108..129)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     457..476
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     716..737
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(783..836)
                     /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"
ORIGIN
        1 taagctgcaa taaacaagtt ggggtgggcg aagaactcca gcatgagatc cccgcgctgc
       61 agttacttgt acagctcgtc catgccgaga gtgatcccgg cggcggtcac gaactccagc
      121 aggaccatgt gatcgcgctt ctcgttgggg tctttgctca gggcggactg ggtgctcagg
      181 tagtggttgt cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtgg
      241 tcggcgagct gcacgctgcc gtcctcgatg ttgtggcgga tcttgaagtt caccttgatg
      301 ccgttcttct gcttgtcggc catgatatag acgttgtggc tgttgtagtt gtactccagc
      361 ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc
      421 accagggtgt cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg
      481 aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc
      541 tgcttcatgt ggtcggggta gcggctgaag cactgcacgc cgtaggtcag ggtggtcacg
      601 agggtgggcc agggcacggg cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg
      661 ccgtaggtgg catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg
      721 ccgtccagct cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc
      781 attgggccgg gattctcctc cacgtcaccg catgttagaa gacttcctct gccctctccg
      841 ccgccggacc tgcccttttt gataatctga gggtgctttt tgctcttcac ctcatacagg
      901 tttcccagaa tgtcggttga gtactttttg atactctgag tcttagaggc gattgttttg
      961 ataattcgag gggg
//
