LOCUS       sequence-121205-         780 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  cloning of sgRNAs for expression in plants.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 780)
  AUTHORS   Ma X, Zhang Q, Zhu Q, Liu W, Chen Y, Qiu R, Wang B, Yang Z, Li H, 
            Lin Y, Xie Y, Shen R, Chen S, Wang Z, Chen Y, Guo J, Chen L, Zhao X,
            Dong Z, Liu YG
  TITLE     A robust CRISPR/Cas9 system for convenient, high-efficiency 
            multiplex genome editing in monocot and dicot plants.
  JOURNAL   Mol Plant. 2015 Apr 24. pii: S1674-2052(15)00204-X. doi: 
            10.1016/j.molp.2015.04.007.
  PUBMED    25917172
REFERENCE   2  (bases 1 to 780)
  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: "Mol Plant. 
            2015 Apr 24. pii: S1674-2052(15)00204-X. doi: 
            10.1016/j.molp.2015.04.007."
COMMENT     Sequence Label: pYLsgRNA-AtU3d
FEATURES             Location/Qualifiers
     source          1..780
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(2..24)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     124..143
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     337..359
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     351..368
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     352..368
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_RNA        complement(571..646)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(696..712)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(696..712)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(709..731)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    720..736
                     /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)."
     promoter        complement(join(744..750,751..768,769..774))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
       61 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
      121 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
      181 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt
      241 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
      301 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
      361 cggccagtgc caagcttagt atgttatctt cagaggtctc ttgaccaatg gtgctttgta
      421 gccatatata aagcagttat ttcttcactc gcttaccgat gtgggacgca aaattcgtaa
      481 gaaaaaagaa atcataagct tattcctttg ctgccgattc cacaggtaaa acggaggaaa
      541 attccatcca ctccaagctc ttgaaaaaaa gcaccgactc ggtgccactt tttcaagttg
      601 ataacggact agccttattt taacttgcta tttctagctc taaaaccgag acccacgctg
      661 gatccccggg taccgagctc gaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat
      721 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
//
