LOCUS       sequence-181904-        3130 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Module B, Promoter: OsU3, Gene: Esp3I ccdb cassette for single gRNA 
            spacer cloning, Terminator: Pol III.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3130)
  AUTHORS   Cermak T, Curtin SJ, Gil-Humanes J, Cegan R, Kono TJY, Konecna E, 
            Belanto JJ, Starker CG, Mathre JW, Greenstein RL, Voytas DF
  TITLE     A multi-purpose toolkit to enable advanced genome engineering in 
            plants.
  JOURNAL   Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 
            10.1105/tpc.16.00922.
  PUBMED    28522548
REFERENCE   2  (bases 1 to 3130)
  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 Cell.
            2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922."
COMMENT     Sequence Label: pMOD_B2519
FEATURES             Location/Qualifiers
     source          1..3130
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      80..668
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     569..588
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     822..839
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        877..1256
                     /label=OsU3 promoter
                     /note="Oryza sativa (rice) snRNA U3 promoter"
     CDS             complement(1264..1569)
                     /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
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(1366..1385)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     misc_RNA        1896..1971
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2038..2142
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gccttgacgg caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
       61 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
      121 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ctaactcttt
      181 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
      241 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
      301 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
      361 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgctcacagc
      421 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
      481 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
      541 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
      601 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
      661 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
      721 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
      781 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
      841 aagcggaaga gcgcccacct gccaggggac ccatggaagg aatctttaaa catacgaaca
      901 gatcacttaa agttcttctg aagcaactta aagttatcag gcatgcatgg atcttggagg
      961 aatcagatgt gcagtcaggg accatagcac aagacaggcg tcttctactg gtgctaccag
     1021 caaatgctgg aagccgggaa cactgggtac gtcggaaacc acgtgatgtg aagaagtaag
     1081 ataaactgta ggagaaaagc atttcgtagt gggccatgaa gcctttcagg acatgtattg
     1141 cagtatgggc cggcccatta cgcaattgga cgacaacaaa gactagtatt agtaccacct
     1201 cggctatcca catagatcaa agctgattta aaagagttgt gcagatgatc cgtggcggag
     1261 acgttatatt ccccagaaca tcaggttaat ggcgtttttg atgtcatttt cgcggtggct
     1321 gagatcagcc acttcttccc cgataacgga aaccggcaca ctggccatat cggtggtcat
     1381 catgcgccag ctttcatccc cgatatgcac caccgggtaa agttcacggg agactttatc
     1441 tgacagcaga cgtgcactgg ccagggggat caccatccgt cgcccgggcg tgtcaataat
     1501 atcactctgt acatccacaa acagacgata acggctctct cttttatagg tgtaaacctt
     1561 aaactgcatt tcaccagccc ctgttctcgt cagcaaaaga gccgttcatt tcaataaacc
     1621 gggcgacctc agccatccct tcctgatttt ccgctttcca gcgttcggca cgcagacgac
     1681 gggcttcatt ctgcatggtt gtgcttacca gaccggagat attgacatca tatatgcctt
     1741 gagcaactga tagctgtcgc tgtcaactgt cactgtaata cgctgcttca tagcatacct
     1801 ccttttgaca tacttcgggt atacatatca gtatatattc ttataccgca aaaatcagcg
     1861 cgcaaatacg catactgtta tctggcttcg tctctgtttt agagctagaa atagcaagtt
     1921 aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg ctttttttcc
     1981 cggtgacgca ggtgatgaaa agccttaggg tcaggtggca cttttcgggg aaatgtgcga
     2041 tgaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct aatgagacaa
     2101 taaccctgat aaatgcttca attatattga aaaaggaaga gtatgagtat ccaacatttc
     2161 cgtgtcgccc ttattccctt ttttgcggcc ttctgccttc ctgtttttgc tcacccagaa
     2221 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cccgagtggg ttacattgaa
     2281 ctggatctca acagcggcaa aatcttagag agttttcgcc ccgaagaacg cttcccaatg
     2341 atgagcactt tcaaagttct gctatgtggc gcggtgttat cccgtattga tgctgggcaa
     2401 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ttcaccagtc
     2461 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgtagtgc tgccataacc
     2521 atgagtgata acactgcggc caacttactt ctgacaacga ttggaggacc gaaggagcta
     2581 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     2641 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     2701 actctccgca aactattaac tggcgaacta cttactcttg cttcacgcca gcaactcatt
     2761 gactggatgg aggcggacaa agttgcagga ccacttctgc gctcggcact tccggctggc
     2821 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     2881 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     2941 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     3001 taactgtcag acgaagggaa ataaatagta gcccgcctga tgtgcgggct tttttttggt
     3061 ggtggttacc agtgtcgctt agctaccgta ggtcctctta ggcggtccga ttagcgacat
     3121 agtcatcaat
//
