LOCUS       sequence-152254-        1031 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Fusion protein Cas9+GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1031)
  AUTHORS   Pulido-Quetglas C, Aparicio-Prat E, Arnan C, Polidori T, Hermoso T, 
            Palumbo E, Ponomarenko J, Guigo R, Johnson R
  TITLE     Scalable Design of Paired CRISPR Guide RNAs for Genomic Deletion.
  JOURNAL   PLoS Comput Biol. 2017 Mar 2;13(3):e1005341. doi: 
            10.1371/journal.pcbi.1005341. eCollection 2017 Mar.
  PUBMED    28253259
REFERENCE   2  (bases 1 to 1031)
  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: "PLoS Comput
            Biol."; date: "2017-03-2"; volume: "13(3):e1005341. doi"; pages: " 
            10.1371/journal.pcbi.1005341. eCollection 2017 Mar"
COMMENT     Sequence Label: pLentiCas9-GFP
FEATURES             Location/Qualifiers
     source          1..1031
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             48..95
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             96..119
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(126..128,129..131,132..842)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKLSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(171..192)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(432..451)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     779..800
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             843..860
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             870..926
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
ORIGIN
        1 ccggcctgta cgagacacgg atcgacctgt ctcagctggg aggcgacaag cgacctgccg
       61 ccacaaagaa ggctggacag gctaagaaga agaaagatta caaagacgat gacgataagg
      121 gatccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
      181 tggacggcga cgtaaacggc cacaagctca gcgtgtccgg cgagggcgag ggcgatgcca
      241 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
      301 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
      361 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
      421 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
      481 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
      541 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
      601 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
      661 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
      721 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
      781 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
      841 agcatcatca ccatcaccac ggatccggcg caacaaactt ctctctgctg aaacaagccg
      901 gagatgtcga agagaatcct ggaccgatgg ccaagccttt gtctcaagaa gaatccaccc
      961 tcattgaaag agcaacggct acaatcaaca gcatccccat ctctgaagac tacagcgtcg
     1021 ccagcgcagc t
//
