LOCUS       sequence-69838-e         853 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  human gRNA expression vector targeting EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 853)
  AUTHORS   Fu Y, Foden JA, Khayter C, Maeder ML, Reyon D, Joung JK, Sander JD
  TITLE     High-frequency off-target mutagenesis induced by CRISPR-Cas 
            nucleases in human cells.
  JOURNAL   Nat Biotechnol. 2013 Jun 23. doi: 10.1038/nbt.2623.
  PUBMED    23792628
REFERENCE   2  (bases 1 to 853)
  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: "Nat 
            Biotechnol. 2013 Jun 23. doi: 10.1038/nbt.2623."
COMMENT     Sequence Label: pFYF1328 EGFP Site#3
FEATURES             Location/Qualifiers
     source          1..853
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     126..145
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        224..299
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(483..502)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aggctgttag agagataatt agaattaatt tgactgtaaa cacaaagata ttagtacaaa
       61 atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta
      121 aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata
      181 tcttgtggaa aggacgaaac accggtggtg cagatgaact tcagttttag agctagaaat
      241 agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct
      301 ttttttaagc ttgggccgct cgaggtacct ctctacatat gacatgtgag caaaaggcca
      361 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
      421 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
      481 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
      541 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
      601 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
      661 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
      721 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
      781 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
      841 aagaacagta ttt
//
