LOCUS       sequence-157816-         893 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Lentiviral construct for building stable cell lines expressing Cas9 
            in the presence of TNFa via puromycin selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 893)
  AUTHORS   Perli SD, Cui CH, Lu TK
  TITLE     Continuous genetic recording with self-targeting CRISPR-Cas in human
            cells.
  JOURNAL   Science. 2016 Aug 18. pii: aag0511.
  PUBMED    27540006
REFERENCE   2  (bases 1 to 893)
  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: "Science. 
            2016 Aug 18. pii: aag0511."
COMMENT     Sequence Label: Construct 33 - NFKBRp_Cas9_3xNLS_p2a-puroR
FEATURES             Location/Qualifiers
     source          1..893
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(460..516)
                     /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"
     CDS             complement(535..555)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(562..582)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(607..654)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
ORIGIN
        1 ggcgcgctcg gccgcctcca ctccggggag cacgacggcg ctgcccagac ccttgccctg
       61 gtggtcgggc gagacgccga cggtggccag gaaccacgcg ggctccttgg gccggtgcgg
      121 cgccaggagg ccttccatct gttgctgcgc ggccagccgg gaaccgctca actcggccat
      181 gcgcgggccg atctcggcga acaccgcccc cgcttcgacg ctctccggcg tggtccagac
      241 cgccaccgcg gcgccgtcgt ccgcgaccca caccttgccg atgtcgagcc cgacgcgcgt
      301 gaggaagagt tcttgcagct cggtgacccg ctcgatgtgg cggtccgggt cgacggtgtg
      361 gcgcgtggcg gggtagtcgg cgaacgcggc ggcgagggtg cgtacggccc gggggacgtc
      421 gtcgcgggtg gcgaggcgca ccgtgggctt gtactcggta ggtccagggt tctcctccac
      481 gtctccagcc tgcttcagca ggctgaagtt agtagctccg cttccagcgc ttatcacctt
      541 cctctttttc ttggggtctt caaccttcct tttcttcttt ggggatccgg cgccgctgcc
      601 gctagctttc tttttcttag cttgaccagc tttcttagta gcagcaggac gcttgtcgcc
      661 tcccagctga gacaggtcga tccgtgtctc gtacaggccg gtgatgctct ggtggatcag
      721 ggtggcgtcc agcacctctt tggtgctggt gtacctcttc cggtcgatgg tggtgtcaaa
      781 gtacttgaag gcggcagggg ctcccagatt ggtcagggta aacaggtgga tgatattctc
      841 ggcctgctct ctgatgggct tatcccggtg cttgttgtag gcggacagca ctt
//
