LOCUS       sequence-142169-        1024 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 1024)
  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 1024)
  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..1024
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    95..116
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(131..136,137..154,155..161)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    169..185
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     174..196
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     193..209
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     193..209
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        395..698
                     /label=AtU6-1 promoter
                     /note="promoter for the Arabidopsis thaliana U6-1 snRNA 
                     gene (Waibel and Filipowicz, 1990)"
     misc_RNA        719..794
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(806..830)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(888..905)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(888..904)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(897..919)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
       61 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
      121 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
      181 aacaatttca cacaggaaac agctatgacc atgggcgcgc caagcttagg cctgtcgacg
      241 cccgggcggt accgcgatcg ctcgcgacct gcaggcataa agccgtcagt gtccgcataa
      301 agaaccaccc ataataccca taatagctgt ttgccatcgc taccttagga ccgttatagt
      361 taattaccct gttatcccta ttaattaaga gctcagaaat ctcaaaattc cggcagaaca
      421 attttgaatc tcgatccgta gaaacgagac ggtcattgtt ttagttccac cacgattata
      481 tttgaaattt acgtgagtgt gagtgagact tgcataagaa aataaaatct ttagttggga
      541 aaaaattcaa taatataaat gggcttgaga aggaagcgag ggataggcct ttttctaaaa
      601 taggcccatt taagctatta acaatcttca aaagtaccac agcgcttagg taaagaaagc
      661 agctgagttt atatatggtt agagacgaag tagtgattgg acttttgcca gccatggtgt
      721 tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg
      781 caccgagtcg gtgctttttt tctagaccca gctttcttgt acaaagttgg cattacccat
      841 cggcgcgcct ctagaattta aatggatcct acgtactcga ggaattcact ggccgtcgtt
      901 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
      961 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     1021 ttgc
//
