LOCUS       sequence-122225-        1303 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Level 1 Golden Gate Cassette: sgRNA cassette targeting BolC.GA4a-1 
            in Brassica oleracea.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1303)
  AUTHORS   Lawrenson T, Shorinola O, Stacey N, Li C, Ostergaard L, Patron N, 
            Uauy C, Harwood W
  TITLE     Induction of targeted, heritable mutations in barley and Brassica 
            oleracea using RNA-guided Cas9 nuclease.
  JOURNAL   Genome Biol. 2015 Nov 30;16(1):258. doi: 10.1186/s13059-015-0826-7.
  PUBMED    26616834
REFERENCE   2  (bases 1 to 1303)
  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: "Genome 
            Biol."; date: "2015-11-30"; volume: "16(1):258. doi"; pages: " 
            10.1186/s13059-015-0826-7"
COMMENT     Sequence Label: pICSL11061
FEATURES             Location/Qualifiers
     source          1..1303
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_RNA        307..382
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(394..418)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    469..493
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
ORIGIN
        1 acnttgcggn gttttaatgt actggggtgg atgcagtggg ccccactctg tgaagacaaa
       61 ctaccccttt gagngncgga gcatcttcat tcttaagata tgaagataat cttcaaaagg
      121 cccctgggaa tctgaaagaa gagaagcagg cccatttata tgggaaagaa caatagtatt
      181 tcttatatag gcccatttaa gttgaaaaca atcttcaaaa gtcccacatc gcttagataa
      241 gaaaacgaag ctgagtttat atacagctag agtcgaagta nggattgttt tcacttgcgg
      301 ccggaggttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga
      361 aaaagtggca ccgagtcggt gctttttttc tagacccagc tttcttgtac aaagttggca
      421 ttacgcttta cttgtcttct gcacgaagtg gtttaaacta ncagtgtttg acaggatata
      481 ttggcgggta aacctaanag aaaagagcgt ttattagaat aatcggatat ttaaaagggc
      541 gtgaaaaggt ttatccgttc gtccatttgt atgtgcatgc caaccacagg gttccccaga
      601 tcaggcgctg gctgctgaac ccccagcngg aactgacccc acnaggccct agcgttngcn
      661 atgccccang tcatcattga cccangcggt gttccaccag gccgctgcct cgcaactctt
      721 cgcanggcta ngccganctg ntcgcgccac ttcttcangg cgggtggant ccnaatccnn
      781 cncangnagg nggaaangtt tccngcttgn aaccnggtac nggncncccg ggtgcnaagc
      841 tggaaaanng tcgaaanatc ncgncgggcc gtcngccnan ggcttggcgg nacctnctcc
      901 cnaagnngaa ntttccgcgt anncgnttnn ccagnanacc gntacnnacn aatttccccn
      961 gtacnancan gnacctggcc acgggnacng ttttnttgcc acgggcccag ganncgaaan
     1021 acnggngntt caacnanacc cgattnctgg gtgcccnaac nnggcccgnc nnncgaatcc
     1081 natcnncnnc cccnngnang ggcnnaangn aattccnggg gcanagtngn anancgggcn
     1141 ngncngcgnn cnccnggntc ccggatanan cggtgaaacg gcaggangaa ccnatcncca
     1201 aanggngagg gttcgntgtt ccnatnnnga ggcccanata ttgtatnang acanngatgc
     1261 gncacggagn ngnaatnana ggcctngnnc aangannnac cct
//
