LOCUS       sequence-81574-e        1034 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Can use a PCR template to assemble new desired guide RNA. Contains 
            an Arabidopsis U6 promoter to drive guide RNA (targeting AtPDS3 gene
            target site 1) expression with a TTTTTT as terminator. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1034)
  AUTHORS   Li JF, Norville JE, Aach J, McCormack M, Zhang D, Bush J, Church GM,
            Sheen J
  TITLE     Multiplex and homologous recombination-mediated genome editing in 
            Arabidopsis and Nicotiana benthamiana using guide RNA and Cas9.
  JOURNAL   Nat Biotechnol. 2013 Aug;31(8):688-91. doi: 10.1038/nbt.2654.
  PUBMED    23929339
REFERENCE   2  (bases 1 to 1034)
  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"; doi: "10.1038/nbt"; 
            journalName: "Nat Biotechnol"; date: "2013-08"; volume: "31"; issue:
            "8"; pages: "688-91"
COMMENT     Sequence Label: pUC119-gRNA
FEATURES             Location/Qualifiers
     source          1..1034
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     9..25
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9..25
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        211..514
                     /label=AtU6-1 promoter
                     /note="promoter for the Arabidopsis thaliana U6-1 snRNA 
                     gene (Waibel and Filipowicz, 1990)"
     misc_RNA        535..610
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(622..646)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(704..721)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(704..720)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(713..735)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 atttcacaca ggaaacagct atgaccatgg gcgcgccaag cttaggcctg tcgacgcccg
       61 ggcggtaccg cgatcgctcg cgacctgcag gcataaagcc gtcagtgtcc gcataaagaa
      121 ccacccataa tacccataat agctgtttgc catcgctacc ttaggaccgt tatagttaat
      181 taccctgtta tccctattaa ttaagagctc agaaatctca aaattccggc agaacaattt
      241 tgaatctcga tccgtagaaa cgagacggtc attgttttag ttccaccacg attatatttg
      301 aaatttacgt gagtgtgagt gagacttgca taagaaaata aaatctttag ttgggaaaaa
      361 attcaataat ataaatgggc ttgagaagga agcgagggat aggccttttt ctaaaatagg
      421 cccatttaag ctattaacaa tcttcaaaag taccacagcg cttaggtaaa gaaagcagct
      481 gagtttatat atggttagag acgaagtagt gattggactt ttgccagcca tggtgtttta
      541 gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc
      601 gagtcggtgc tttttttcta gacccagctt tcttgtacaa agttggcatt acccatcggc
      661 gcgcctctag aatttaaatg gatcctacgt actcgaggaa ttcactggcc gtcgttttac
      721 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
      781 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
      841 gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
      901 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
      961 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     1021 tcctttcgct ttct
//
