LOCUS       sequence-152701-        1117 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  sgRNA shuttle vector; module 2E.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1117)
  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 1117)
  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: pDGE8
FEATURES             Location/Qualifiers
     source          1..1117
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_RNA        128..203
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(283..299)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(283..299)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(296..318)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    307..323
                     /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(331..337,338..355,356..361))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    376..397
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(514..531)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(765..784)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtttccgtta tcggggaaga agtggctgat ctcagccacc gcgaaaatga catcaaaaac
       61 gccattaacc tgatgttctg gggaatataa atgtcaggct cccttataca cagccagtcg
      121 aagacttgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg
      181 aaaaagtggc accgagtcgg tgcttttttt tgcttagaga cctttatcgg atcccgggcc
      241 cgtcgactgc agaggcctgc atgcaagctt ggcgtaatca tggtcatagc tgtttcctgt
      301 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
      361 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
      421 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
      481 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
      541 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
      601 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
      661 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
      721 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
      781 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
      841 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
      901 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
      961 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     1021 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     1081 tacagagttc ttgaagtggt ggcctaacta cggctac
//
