LOCUS       sequence-157791-         946 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 946)
  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 946)
  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 3 - U6p_20nt1_mod2_sgRNA (stgRNA)
FEATURES             Location/Qualifiers
     source          1..946
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(87..143)
                     /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(153..863,864..866,867..869))
                     /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     803..824
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      866..875
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 tccgagagct gcatcaggtc ggagacgctg tcgaactttt cgatcagaaa cttctcgaca
       61 gacgtcgcgg tgagttcagg ctttttaggt ccagggttct cctccacgtc tccagcctgc
      121 ttcagcaggc tgaagttagt agctccgctt cccttgtaca gctcgtccat gccgagagtg
      181 atcccggcgg cggtgcggaa ctccagcagg accatgtgat cgcgcttctc gttggggtct
      241 ttgctcagca cggactgggt gctcaggtag tggctgtcgg gcagcagcac ggggccgtcg
      301 ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc ctccacgttg
      361 tggcggatct tgaagttcac cttgatgccg ttcttctgct tgacggccat gatatagatg
      421 ttgtggctgt tgaagttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag
      481 tcgacgccct tcagctcgat gcggttcact agggtgtcgc cctcgaactt cacctcggcg
      541 cgggtcttgt aggtgccgtc gtccttgaag aagatggtgc gctcctggac gtagccttcg
      601 ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg ggcgaagcac
      661 tgcacgccgt ggctcagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg
      721 gtgcagatga acttcagggt cagcttgccg ttggtggcat cgccctcgcc ctcgcccctc
      781 acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg
      841 gtgaacagct cctcgccctt gctcaccatg gtggcaagct tgggatgcag gtcgaaaggc
      901 ccggagatga ggaagaggag aacagcgcgg cagacgtgcg cttttg
//
