LOCUS       sequence-69837-e         947 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 947)
  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 947)
  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: pFYF1327 EGFP Site#2
FEATURES             Location/Qualifiers
     source          1..947
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     125..144
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        223..298
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(482..501)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggctgttaga gagataatta gaattaattt gactgtaaac acaaagatat tagtacaaaa
       61 tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa
      121 aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat
      181 cttgtggaaa ggacgaaaca ccgatgccgt tcttctgctt gtgttttaga gctagaaata
      241 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
      301 tttttaagct tgggccgctc gaggtacctc tctacatatg acatgtgagc aaaaggccag
      361 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
      421 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
      481 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
      541 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
      601 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
      661 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
      721 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
      781 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
      841 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
      901 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttt
//
