LOCUS       sequence-153869-         779 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  3rd generation lentiviral gRNA plasmid targeting EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 779)
  AUTHORS   Doench JG, Fusi N, Sullender M, Hegde M, Vaimberg EW, Donovan KF, 
            Smith I, Tothova Z, Wilen C, Orchard R, Virgin HW, Listgarten J, 
            Root DE
  TITLE     Optimized sgRNA design to maximize activity and minimize off-target 
            effects of CRISPR-Cas9.
  JOURNAL   Nat Biotechnol. 2016 Jan 18. doi: 10.1038/nbt.3437.
  PUBMED    26780180
REFERENCE   2  (bases 1 to 779)
  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. 2016 Jan 18. doi: 10.1038/nbt.3437."
COMMENT     Sequence Label: EGFP gRNA (BRDN0000562805)
FEATURES             Location/Qualifiers
     source          1..779
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_RNA        complement(58..133)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(155..403)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(213..232)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(383..403)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     CDS             complement(581..625)
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
ORIGIN
        1 aattgtggat gaatactgcc atttgtctca agatctagtt acgccaagct taaaaaagca
       61 ccgactcggt gccacttttt caagttgata acggactagc cttattttaa cttgctattt
      121 ctagctctaa aacgctcgac caggatgggc acccggtgtt tcgtcctttc cacaagatat
      181 ataaagccaa gaaatcgaaa tactttcaag ttacggtaag catatgatag tccattttaa
      241 aacataattt taaaactgca aactacccaa gaaattatta ctttctacgt cacgtatttt
      301 gtactaatat ctttgtgttt acagtcaaat taattctaat tatctctcta acagccttgt
      361 atcgtatatg caaatatgaa ggaatcatgg gaaataggcc ctctctgggt cccctcgggg
      421 ttgggaggtg ggtctgaaac gataatggtg aatatccctg cctaactcta ttcactatag
      481 aaagtacagc aaaaactatt cttaaaccta ccaagcctcc tactatcatt atgaataatt
      541 ttatatacca cagccaattt gttatgttaa accaattcca caaacttgcc catttatcta
      601 attccaataa ttcttgttca ttcttttctt gctggttttg cgattcttca attaaggagt
      661 gtattaagct tgtgtaattg ttaatttctc tgtcccactc catccaggtc gtgtgattcc
      721 aaatctgttc cagagattta ttactccaac tagcattcca aggcacagca gtggtgcaa
//
