LOCUS       sequence-86046-e        1047 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Transcription of guide RNA (gRNA) in wheat cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1047)
  AUTHORS   Shan Q, Wang Y, Li J, Zhang Y, Chen K, Liang Z, Zhang K, Liu J, Xi 
            JJ, Qiu JL, Gao C
  TITLE     Targeted genome modification of crop plants using a CRISPR-Cas 
            system.
  JOURNAL   Nat Biotechnol. 2013 Aug;31(8):686-8. doi: 10.1038/nbt.2650.
  PUBMED    23929338
REFERENCE   2  (bases 1 to 1047)
  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: "686-8"
COMMENT     Sequence Label: pU6-gRNA
FEATURES             Location/Qualifiers
     source          1..1047
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     96..113
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    230..251
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(266..271,272..289,290..296)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    304..320
                     /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     309..331
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     328..344
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     328..344
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_RNA        complement(379..454)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(848..865)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(848..864)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(857..879)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
       61 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
      121 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
      181 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
      241 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
      301 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgaattcg
      361 agctcggtac caaaaaaagc accgactcgg tgccactttt tcaagttgat aacggactag
      421 ccttatttta acttgctatt tctagctcta aaacgtgtct tcttgtgttg aagacagcaa
      481 gtctgatgca gcaagcgagt ttataaaccg gtcagaactc tgctggacag gcgaggtggg
      541 actaaactaa gaggaactgc tgcgttgctc tccccaatcg tgaacacgtc gaccacccga
      601 caaagcccga tcatgcgggc ctaaaagccc agcgctagac cgttgctgaa ctcacgtggg
      661 ctgcatctca ctatgcgccc agatcagtga gagctagagg gcagttatct tcagctctct
      721 cgtagttcct ctgatcaata ttcctagttc gattccctcc tagcagtgag taacaatgtg
      781 ctccttgata aatataaatg tgttgagcac cagaactgtc agaataacgg gcttggtcaa
      841 gcttggcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
      901 ttaatcgcct tgcagcacat cccccctttc gccagctggc gtaatagcga agaggcccgc
      961 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat
     1021 tttctcctta cgcatctgtg cggtatt
//
