LOCUS       sequence-159191-         967 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  guide RNA (gRNA) expression vector used to create a gRNA to a 
            specific sequence; uses T7 promoter for in vitro transcription.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 967)
  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 967)
  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: DR274
FEATURES             Location/Qualifiers
     source          1..967
                     /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"
     promoter        complement(240..258)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(387..403)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(387..403)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      451..482
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        483..585
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(659..678)
                     /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 tctaaaactg agaccctctc tcggtctctc
      241 ctatagtgag tcgtattagc tagcggtgcg agcggatcga gcagtgtcga tcactactgg
      301 accgcgagct gtgctgcgac ccgtgatctt acggcattat acgtatgatc ggtccacgat
      361 cagctagatt atctagtcag cttgatgtca tagctgtttc ctgaggctca atactgacca
      421 tttaaatcat acctgacctc catagcagaa agtcaaaagc ctccgaccgg aggcttttga
      481 cttgatcggc acgtaagagg ttccaacttt caccataatg aaataagatc actaccgggc
      541 gtattttttg agttatcgag attttcagga gctaaggaag ctaaaatgag ccatattcaa
      601 cgggaaacgt cttgcttgaa gccgcgatta aattccaaca tggatgctga tttatatggg
      661 tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg attgtatggg
      721 aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt
      781 acagatgaga tggtcaggct aaactggctg acggaattta tgcctcttcc gaccatcaag
      841 cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc agggaaaaca
      901 gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca
      961 gtgttcc
//
