LOCUS       sequence-86641-e         806 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Co-expresses human optimized S. pyogenes dCas9 and GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 806)
  AUTHORS   Kabadi AM, Ousterout DG, Hilton IB, Gersbach CA
  TITLE     Multiplex CRISPR/Cas9-based genome engineering from a single 
            lentiviral vector.
  JOURNAL   Nucleic Acids Res. 2014 Aug 13. pii: gku749.
  PUBMED    25122746
REFERENCE   2  (bases 1 to 806)
  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: "Nucleic 
            Acids Res. 2014 Aug 13. pii: gku749."
COMMENT     Sequence Label: pLV hUbC-dCas9-T2A-GFP
FEATURES             Location/Qualifiers
     source          1..806
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    42..75
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(91..801,802..804))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             complement(join(91..801,802..804))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(133..154)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     482..501
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     741..762
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 taatccagag gttgattatc gataagcttg atatcgaatt cataacttcg tataatgtat
       61 gctatacgaa gttatctatt atcaaccggt cttgtacagc tcgtccatgc cgagagtgat
      121 cccggcggcg gtcacgaact ccagcaggac catgtgatcg cgcttctcgt tggggtcttt
      181 gctcagggcg gactgggtgc tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc
      241 gatgggggtg ttctgctggt agtggtcggc gagctgcacg ctgccgtcct cgatgttgtg
      301 gcggatcttg aagttcacct tgatgccgtt cttctgcttg tcggccatga tatagacgtt
      361 gtggctgttg tagttgtact ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc
      421 gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg
      481 ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt agccttcggg
      541 catggcggac ttgaagaagt cgtgctgctt catgtggtcg gggtagcggc tgaagcactg
      601 cacgccgtag gtcagggtgg tcacgagggt gggccagggc acgggcagct tgccggtggt
      661 gcagatgaac ttcagggtca gcttgccgta ggtggcatcg ccctcgccct cgccggacac
      721 gctgaacttg tggccgttta cgtcgccgtc cagctcgacc aggatgggca ccaccccggt
      781 gaacagctcc tcgcccttgc tcacgg
//
