LOCUS       sequence-57632-e         830 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  gRNA targeted to zebrafish gene gsk3b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 830)
  AUTHORS   Hwang WY, Fu Y, Reyon D, Maeder ML, Tsai SQ, Sander JD, Peterson RT,
            Yeh JR, Joung JK
  TITLE     Efficient genome editing in zebrafish using a CRISPR-Cas system.
  JOURNAL   Nat Biotechnol. 2013 Jan 29. doi: 10.1038/nbt.2501.
  PUBMED    23360964
REFERENCE   2  (bases 1 to 830)
  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 Jan 29. doi: 10.1038/nbt.2501."
COMMENT     Sequence Label: Zebrafish-gRNA-0005
FEATURES             Location/Qualifiers
     source          1..830
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_RNA        complement(143..218)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(236..255)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(237..255)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(384..400)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(384..400)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      448..479
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        480..582
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(656..675)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 aggtgagcca gtgagttgat tgcagtccag ttacgctgga gtctgaggct cgtcctgaat
       61 gatatgcgac cgccggaggg ttgcgtttga gacgggcgac agatccagtc gcgctgctct
      121 cgtcgatcca agctttttaa aagcaccgac tcggtgccac tttttcaagt tgataacgga
      181 ctagccttat tttaacttgc tatttctagc tctaaaaccc tgcggccggt caggtcccta
      241 tagtgagtcg tattagctag cggtgcgagc ggatcgagca gtgtcgatca ctactggacc
      301 gcgagctgtg ctgcgacccg tgatcttacg gcattatacg tatgatcggt ccacgatcag
      361 ctagattatc tagtcagctt gatgtcatag ctgtttcctg aggctcaata ctgaccattt
      421 aaatcatacc tgacctccat agcagaaagt caaaagcctc cgaccggagg cttttgactt
      481 gatcggcacg taagaggttc caactttcac cataatgaaa taagatcact accgggcgta
      541 ttttttgagt tatcgagatt ttcaggagct aaggaagcta aaatgagcca tattcaacgg
      601 gaaacgtctt gcttgaagcc gcgattaaat tccaacatgg atgctgattt atatgggtat
      661 aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag
      721 cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca
      781 gatgagatgg tcaggctaaa ctggctgacg gaatttatgc ctcttccgac
//
