LOCUS       sequence-157786-         871 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  For assaying self-targeting activity via Cas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 871)
  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 871)
  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 1 - U6p_20nt1_WT_sgRNA
FEATURES             Location/Qualifiers
     source          1..871
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(83..139)
                     /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(join(149..859,860..862,863..865))
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     799..820
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      862..871
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 agagctgcat caggtcggag acgctgtcga acttttcgat cagaaacttc tcgacagacg
       61 tcgcggtgag ttcaggcttt ttaggtccag ggttctcctc cacgtctcca gcctgcttca
      121 gcaggctgaa gttagtagct ccgcttccct tgtacagctc gtccatgccg agagtgatcc
      181 cggcggcggt gcggaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc
      241 tcagcacgga ctgggtgctc aggtagtggc tgtcgggcag cagcacgggg ccgtcgccga
      301 tgggggtgtt ctgctggtag tggtcggcga gctgcacgct gccgtcctcc acgttgtggc
      361 ggatcttgaa gttcaccttg atgccgttct tctgcttgac ggccatgata tagatgttgt
      421 ggctgttgaa gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga
      481 cgcccttcag ctcgatgcgg ttcactaggg tgtcgccctc gaacttcacc tcggcgcggg
      541 tcttgtaggt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca
      601 tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcgggcg aagcactgca
      661 cgccgtggct cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc
      721 agatgaactt cagggtcagc ttgccgttgg tggcatcgcc ctcgccctcg cccctcacgc
      781 tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga
      841 acagctcctc gcccttgctc accatggtgg c
//
