LOCUS       sequence-152709-        1112 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  sgRNA shuttle vector; module 5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1112)
  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 1112)
  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: pDGE12
FEATURES             Location/Qualifiers
     source          1..1112
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_RNA        129..204
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(281..297)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(281..297)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(294..316)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    305..321
                     /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(329..335,336..353,354..359))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    374..395
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(512..529)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(763..782)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggtttccgtt atcggggaag aagtggctga tctcagccac cgcgaaaatg acatcaaaaa
       61 cgccattaac ctgatgttct ggggaatata aatgtcaggc tcccttatac acagccagtc
      121 gaagacttgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt
      181 gaaaaagtgg caccgagtcg gtgctttttt tttccgtgag accatcggat cccgggcccg
      241 tcgactgcag aggcctgcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt
      301 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
      361 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
      421 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
      481 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
      541 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
      601 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
      661 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
      721 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
      781 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
      841 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
      901 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
      961 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     1021 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     1081 cagagttctt gaagtggtgg cctaactacg gc
//
