LOCUS       sequence-131997-         838 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Used to construct a gRNA expression cassette for CRISPR/Cas9 genome 
            editing in Marchantia polymorpha.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 838)
  AUTHORS   Sugano SS, Nishihama R, Shirakawa M, Takagi J, Matsuda Y, Ishida S, 
            Shimada T, Hara-Nishimura I, Osakabe K, Kohchi T
  TITLE     Efficient CRISPR/Cas9-based genome editing and its application to 
            conditional genetic analysis in Marchantia polymorpha.
  JOURNAL   PLoS One. 2018 Oct 31;13(10):e0205117. doi: 
            10.1371/journal.pone.0205117. eCollection 2018.
  PUBMED    30379827
REFERENCE   2  (bases 1 to 838)
  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: "PLoS One.";
            date: "2018-10-31"; volume: "13(10):e0205117. doi"; pages: " 
            10.1371/journal.pone.0205117. eCollection 2018"
COMMENT     Sequence Label: pMpGE_En01
FEATURES             Location/Qualifiers
     source          1..838
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    73..97
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        complement(124..199)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             complement(237..542)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(339..358)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
ORIGIN
        1 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
       61 tttcttataa tgccaacttt gtacaagaaa gctgggtcgg cgcgcccacc ctttagaaaa
      121 aaagcaccga ctcggtgcca ctttttcaag ttgataacgg actagcctta ttttaacttg
      181 ctatttctag ctctaaaact gcaggcagac tggctgtgta taagggagcc tgacatttat
      241 attccccaga acatcaggtt aatggcgttt ttgatgtcat tttcgcggtg gctgagatca
      301 gccacttctt ccccgataac ggagaccggc acactggcca tatcggtggt catcatgcgc
      361 cagctttcat ccccgatatg caccaccggg taaagttcac gggggacttt atctgacagc
      421 agacgtgcac tggccagggg gatcaccatc cgtcgcccgg gcgtgtcaat aatatcactc
      481 tgtacatcca caaacagacg ataacggctc tctcttttat aggtgtaaac cttaaactgc
      541 atttcaccag cccctgttct cgtcagcaaa agagccgttc atttcaataa accgggcgac
      601 ctcagccatc ccttcctgat tttccgcttt ccagcgttcg gcacgcagac gacgggcttc
      661 attctgcatg gttgtgctta ccagaccgga gatattgaca tcatatatgc cttgagcaac
      721 tgatagctgt cgctgtcaac tgtcactgta atacgctgct tcatagcata cctctttttg
      781 acatacttcg ggtatacata tcagtatata ttcttatacc gcaaaaatca gcgcgcaa
//
