LOCUS       sequence-152703-        1004 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  sgRNA shuttle vector; module 3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1004)
  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 1004)
  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: pDGE9
FEATURES             Location/Qualifiers
     source          1..1004
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_RNA        123..198
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(278..294)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(278..294)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(291..313)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    302..318
                     /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(326..332,333..350,351..356))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    371..392
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(509..526)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(760..779)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttatcggg gaagaagtgg ctgatctcag ccaccgcgaa aatgacatca aaaacgccat
       61 taacctgatg ttctggggaa tataaatgtc aggctccctt atacacagcc agtcgaagac
      121 ttgttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa
      181 gtggcaccga gtcggtgctt ttttttatgg tgagaccaaa atcggatccc gggcccgtcg
      241 actgcagagg cctgcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
      301 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
      361 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
      421 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
      481 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
      541 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
      601 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
      661 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
      721 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
      781 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
      841 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
      901 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
      961 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggta
//
