LOCUS       sequence-112224-         973 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 973)
  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 973)
  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..973
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(31..133)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(134..165)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     primer_bind     213..229
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     213..229
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        358..376
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        398..473
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(655..672)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(655..671)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gcaagacgtt tcccgttgaa tatggctcat tttagcttcc ttagctcctg aaaatctcga
       61 taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct
      121 tacgtgccga tcaagtcaaa agcctccggt cggaggcttt tgactttctg ctatggaggt
      181 caggtatgat ttaaatggtc agtattgagc ctcaggaaac agctatgaca tcaagctgac
      241 tagataatct agctgatcgt ggaccgatca tacgtataat gccgtaagat cacgggtcgc
      301 agcacagctc gcggtccagt agtgatcgac actgctcgat ccgctcgcac cgctagctaa
      361 tacgactcac tataggagag accgagagag ggtctcagtt ttagagctag aaatagcaag
      421 ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttaaa
      481 aagcttggat cgacgagagc agcgcgactg gatctgtcgc ccgtctcaaa cgcaaccctc
      541 cggcggtcgc atatcattca ggacgagcct cagactccag cgtaactgga ctgcaatcaa
      601 ctcactggct caccttccgg tccacgatca gctagaatca agctgactag ataaactggc
      661 cgtcgtttta cacgggtggg cctttcttcg gtagaaaatc aaaggatctt cttgagatcc
      721 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
      781 ttgtttgccg gatcaagagc taccaactct ttttccgagg taactggctt cagcagagcg
      841 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
      901 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
      961 gataagtcgt gtc
//
