LOCUS       sequence-152468-        1118 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  sgRNA shuttle vector; module 1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1118)
  AUTHORS   Ordon J, Gantner J, Kemna J, Schwalgun L, Reschke M, Streubel J, 
            Boch J, Stuttmann J
  TITLE     Generation of chromosomal deletions in dicotyledonous plants 
            employing a user-friendly genome editing toolkit.
  JOURNAL   Plant J. 2016 Aug 31. doi: 10.1111/tpj.13319.
  PUBMED    27579989
REFERENCE   2  (bases 1 to 1118)
  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 J. 
            2016 Aug 31. doi: 10.1111/tpj.13319."
COMMENT     Sequence Label: pDGE5
FEATURES             Location/Qualifiers
     source          1..1118
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_RNA        130..205
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(284..300)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(284..300)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(297..319)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    308..324
                     /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(332..338,339..356,357..362))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    377..398
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(515..532)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(766..785)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cggtttccgt tatcggggaa gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa
       61 acgccattaa cctgatgttc tggggaatat aaatgtcagg ctcccttata cacagccagt
      121 cgaagacttg ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact
      181 tgaaaaagtg gcaccgagtc ggtgcttttt tttcccttga gaccaaatcg gatcccgggc
      241 ccgtcgactg cagaggcctg catgcaagct tggcgtaatc atggtcatag ctgtttcctg
      301 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
      361 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
      421 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
      481 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
      541 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
      601 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
      661 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
      721 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
      781 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
      841 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
      901 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
      961 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     1021 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     1081 ctacagagtt cttgaagtgg tggcctaact acggctac
//
