LOCUS       sequence-100826-         858 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  for generation of a eGFP specific sgRNA from a T7 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 858)
  AUTHORS   Auer TO, Duroure K, De Cian A, Concordet JP, Del Bene F
  TITLE     Highly efficient CRISPR/Cas9-mediated knock-in in zebrafish by 
            homology-independent DNA repair.
  JOURNAL   Genome Res. 2014 Jan;24(1):142-53. doi: 10.1101/gr.161638.113. Epub 
            2013 Oct 31.
  PUBMED    24179142
REFERENCE   2  (bases 1 to 858)
  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"; doi: 
            "10.1101/gr.161638.113"; journalName: "Genome Res"; date: "2014-01";
            volume: "24"; issue: "1"; pages: "142-53"
COMMENT     Sequence Label: DR274-eGFP sgRNA
FEATURES             Location/Qualifiers
     source          1..858
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_RNA        complement(160..235)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(254..272)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(401..417)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(401..417)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      465..496
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        497..599
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(673..692)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 gctgatcgtg gaccggaagg tgagccagtg agttgattgc agtccagtta cgctggagtc
       61 tgaggctcgt cctgaatgat atgcgaccgc cggagggttg cgtttgagac gggcgacaga
      121 tccagtcgcg ctgctctcgt cgatccaagc tttttaaaag caccgactcg gtgccacttt
      181 ttcaagttga taacggacta gccttatttt aacttgctat ttctagctct aaaactaggt
      241 ggcatcgccc tcgcctatag tgagtcgtat tagctagcgg tgcgagcgga tcgagcagtg
      301 tcgatcacta ctggaccgcg agctgtgctg cgacccgtga tcttacggca ttatacgtat
      361 gatcggtcca cgatcagcta gattatctag tcagcttgat gtcatagctg tttcctgagg
      421 ctcaatactg accatttaaa tcatacctga cctccatagc agaaagtcaa aagcctccga
      481 ccggaggctt ttgacttgat cggcacgtaa gaggttccaa ctttcaccat aatgaaataa
      541 gatcactacc gggcgtattt tttgagttat cgagattttc aggagctaag gaagctaaaa
      601 tgagccatat tcaacgggaa acgtcttgct tgaagccgcg attaaattcc aacatggatg
      661 ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct
      721 atcgattgta tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg
      781 ttgccaatga tgttacagat gagatggtca ggctaaactg gctgacggaa tttatgcctc
      841 ttccgaccat caagcatt
//
