LOCUS       sequence-152266-        1117 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Cas9 protein with BFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1117)
  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 1117)
  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-T2A-BFP
FEATURES             Location/Qualifiers
     source          1..1117
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             53..100
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             101..124
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             134..187
                     /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             200..898
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     CDS             908..964
                     /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 catcaccggc ctgtacgaga cacggatcga cctgtctcag ctgggaggcg acaagcgacc
       61 tgccgccaca aagaaggctg gacaggctaa gaagaagaaa gattacaaag acgatgacga
      121 taagggatcc ggagagggca gaggaagtct gctaacatgc ggtgacgtcg aggagaatcc
      181 tggcccaccg gtcgccacca tgagcgagct gattaaggag aacatgcaca tgaagctgta
      241 catggagggc accgtggaca accatcactt caagtgcaca tccgagggcg aaggcaagcc
      301 ctacgagggc acccagacca tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc
      361 cttcgacatc ctggctacta gcttcctcta cggcagcaag accttcatca accacaccca
      421 gggcatcccc gacttcttca agcagtcctt ccctgagggc ttcacatggg agagagtcac
      481 cacatacgaa gacgggggcg tgctgaccgc tacccaggac accagcctcc aggacggctg
      541 cctcatctac aacgtcaaga tcagaggggt gaacttcaca tccaacggcc ctgtgatgca
      601 gaagaaaaca ctcggctggg aggccttcac cgagacgctg taccccgctg acggcggcct
      661 ggaaggcaga aacgacatgg ccctgaagct cgtgggcggg agccatctga tcgcaaacat
      721 caagaccaca tatagatcca agaaacccgc taagaacctc aagatgcctg gcgtctacta
      781 tgtggactac agactggaaa gaatcaagga ggccaacaac gagacctacg tcgagcagca
      841 cgaggtggca gtggccagat actgcgacct ccctagcaaa ctggggcaca agcttaatgg
      901 atccggcgca acaaacttct ctctgctgaa acaagccgga gatgtcgaag agaatcctgg
      961 accgatggcc aagcctttgt ctcaagaaga atccaccctc attgaaagag caacggctac
     1021 aatcaacagc atccccatct ctgaagacta cagcgtcgcc agcgcagctc tctctagcga
     1081 cggccgcatc ttcactggtg tcaatgtata tcatttt
//
