LOCUS       sequence-153867-         778 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 778)
  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 778)
  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 (BRDN0000561167)
FEATURES             Location/Qualifiers
     source          1..778
                     /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(154..402)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(212..231)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(382..402)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     CDS             complement(580..624)
                     /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 aaccggtgaa cagctcctcg cccggtgttt cgtcctttcc acaagatata
      181 taaagccaag aaatcgaaat actttcaagt tacggtaagc atatgatagt ccattttaaa
      241 acataatttt aaaactgcaa actacccaag aaattattac tttctacgtc acgtattttg
      301 tactaatatc tttgtgttta cagtcaaatt aattctaatt atctctctaa cagccttgta
      361 tcgtatatgc aaatatgaag gaatcatggg aaataggccc tctctgggtc ccctcggggt
      421 tgggaggtgg gtctgaaacg ataatggtga atatccctgc ctaactctat tcactataga
      481 aagtacagca aaaactattc ttaaacctac caagcctcct actatcatta tgaataattt
      541 tatataccac agccaatttg ttatgttaaa ccaattccac aaacttgccc atttatctaa
      601 ttccaataat tcttgttcat tcttttcttg ctggttttgc gattcttcaa ttaaggagtg
      661 tattaagctt gtgtaattgt taatttctct gtcccactcc atccaggtcg tgtgattcca
      721 aatctgttcc agagatttat tactccaact agcattccaa ggcacagcag tggtgcaa
//
