LOCUS       sequence-78313-e         960 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Level 2 construct.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 960)
  AUTHORS   Belhaj K, Chaparro-Garcia A, Kamoun S, Nekrasov V
  TITLE     Plant genome editing made easy: targeted mutagenesis in model and 
            crop plants using the CRISPR/Cas system.
  JOURNAL   Plant Methods. 2013 Oct 11;9(1):39.
  PUBMED    24112467
REFERENCE   2  (bases 1 to 960)
  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: "Plant 
            Methods."; date: "2013-10-11"; volume: "9(1)"; pages: "39"
COMMENT     Sequence Label: 
            pAGM4723::AtU6p::sgRNA2-2x35S-5′UTR::Cas9::NOST-AtU6p::sgRNA1
FEATURES             Location/Qualifiers
     source          1..960
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_feature    191..215
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        304..378
                     /label=AtU6 promoter
                     /note="consensus promoter from the three Arabidopsis U6 
                     snRNA genes (Waibel and Filipowicz, 1990; Nekrasov et al., 
                     2013)"
     misc_RNA        399..474
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(486..510)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
       61 cagcaacgcg gcctttttac ggttcctgct cggatctgtt ggaccggaca gtagtcatgg
      121 ttgatgggct gcctgtatcg agtggtgatt ttgtgccgag ctgccggtcg gggagctgtt
      181 ggctggctgg tggcaggata tattgtggtg taaacaaatt gacgcttaga caacttaata
      241 acacattgcg gacgttttta atgtactggg gttgaacact ctgtgccgaa ttcggatccg
      301 gagtgatcaa aagtcccaca tcgatcaggt gatatatagc agcttagttt atataatgat
      361 agagtcgaca tagcgattgt caagatgttt gcttgcaagt tttagagcta gaaatagcaa
      421 gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt
      481 tctagaccca gctttcttgt acaaagttgg cattacgctg caagaattca agcttggagg
      541 tcaacatggt ggagcacgac actctggtct actccaaaaa tgtcaaagat acagtctcag
      601 aagatcaaag ggctattgag acttttcaac aaaggataat ttcgggaaac ctcctcggat
      661 tccattgccc agctatctgt cacttcatcg aaaggacagt agaaaaggaa ggtggctcct
      721 acaaatgcca tcattgcgat aaaggaaagg ctatcattca agatctctct gccgacagtg
      781 gtcccaaaga tggaccccca cccacgagga gcatcgtgga aaaagaagag gttccaacca
      841 cgtctacaaa gcaagtggat tgatgtgata acatggtgga gcacgacact ctggtctact
      901 ccaaaaatgt caaagataca gtctcagaag atcaaagggc tattgagact tttcaacaaa
//
