LOCUS       sequence-95672-e         665 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 665)
  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 665)
  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..665
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        38..112
                     /label=AtU6 promoter
                     /note="consensus promoter from the three Arabidopsis U6 
                     snRNA genes (Waibel and Filipowicz, 1990; Nekrasov et al., 
                     2013)"
     misc_RNA        133..208
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(220..244)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    313..337
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
ORIGIN
        1 gttactagat cgacgctact agaattcgag ctcggagtga tcaaaagtcc cacatcgatc
       61 aggtgatata tagcagctta gtttatataa tgatagagtc gacatagcga ttgccgttaa
      121 tttgagagtc cagttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca
      181 acttgaaaaa gtggcaccga gtcggtgctt tttttctaga cccagctttc ttgtacaaag
      241 ttggcattac gctttacgag gatgcacatg tgaccgaggg acacgaagtg atccgtttaa
      301 actatcagtg tttgacagga tatattggcg ggtaaaccta agagaaaaga gcgtttatta
      361 gaataatcgg atatttaaaa gggcgtgaaa aggtttatcc gttcgtccat ttgtatgtgc
      421 cagccgtgcg gctgcatgaa atcctggccg gtttgtctga tgccaagctg gcggcctggc
      481 cggccagctt ggccgctgaa gaaaccgagc gccgccgtct aaaaaggtga tgtgtatttg
      541 agtaaaacag cttgcgtcat gcggtcgctg cgtatatgat gcgatgagta aataaacaaa
      601 tacgcaaggg gaacgcatga aggttatcgc tgtacttaac cagaaaggcg ggtcaggcaa
      661 gacga
//
