LOCUS       sequence-182053-        3202 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Module C, Promoter: OsU3, Gene: Esp3I ccdb cassette for single gRNA 
            spacer cloning (+ BaeI site for GT donor cloning), Terminator: Pol 
            III.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3202)
  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 3202)
  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_C2519
FEATURES             Location/Qualifiers
     source          1..3202
                     /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        879..1258
                     /label=OsU3 promoter
                     /note="Oryza sativa (rice) snRNA U3 promoter"
     CDS             complement(1266..1571)
                     /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(1368..1387)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     misc_RNA        1898..1973
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2110..2214
                     /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 gccaggccgg acccatggaa ggaatcttta aacatacgaa
      901 cagatcactt aaagttcttc tgaagcaact taaagttatc aggcatgcat ggatcttgga
      961 ggaatcagat gtgcagtcag ggaccatagc acaagacagg cgtcttctac tggtgctacc
     1021 agcaaatgct ggaagccggg aacactgggt acgtcggaaa ccacgtgatg tgaagaagta
     1081 agataaactg taggagaaaa gcatttcgta gtgggccatg aagcctttca ggacatgtat
     1141 tgcagtatgg gccggcccat tacgcaattg gacgacaaca aagactagta ttagtaccac
     1201 ctcggctatc cacatagatc aaagctgatt taaaagagtt gtgcagatga tccgtggcgg
     1261 agacgttata ttccccagaa catcaggtta atggcgtttt tgatgtcatt ttcgcggtgg
     1321 ctgagatcag ccacttcttc cccgataacg gaaaccggca cactggccat atcggtggtc
     1381 atcatgcgcc agctttcatc cccgatatgc accaccgggt aaagttcacg ggagacttta
     1441 tctgacagca gacgtgcact ggccaggggg atcaccatcc gtcgcccggg cgtgtcaata
     1501 atatcactct gtacatccac aaacagacga taacggctct ctcttttata ggtgtaaacc
     1561 ttaaactgca tttcaccagc ccctgttctc gtcagcaaaa gagccgttca tttcaataaa
     1621 ccgggcgacc tcagccatcc cttcctgatt ttccgctttc cagcgttcgg cacgcagacg
     1681 acgggcttca ttctgcatgg ttgtgcttac cagaccggag atattgacat catatatgcc
     1741 ttgagcaact gatagctgtc gctgtcaact gtcactgtaa tacgctgctt catagcatac
     1801 ctccttttga catacttcgg gtatacatat cagtatatat tcttataccg caaaaatcag
     1861 cgcgcaaata cgcatactgt tatctggctt cgtctctgtt ttagagctag aaatagcaag
     1921 ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttt
     1981 cgcgcgtagt cctcggtaat atcgcagaac aaaagtacct gatatcgagt gtacttcaag
     2041 tcacaccggc gagtgactag caggtgatga aaagccttag ggtcaggtgg cacttttcgg
     2101 ggaaatgtgc gatgaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     2161 ctaatgagac aataaccctg ataaatgctt caattatatt gaaaaaggaa gagtatgagt
     2221 atccaacatt tccgtgtcgc ccttattccc ttttttgcgg ccttctgcct tcctgttttt
     2281 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcccgagtg
     2341 ggttacattg aactggatct caacagcggc aaaatcttag agagttttcg ccccgaagaa
     2401 cgcttcccaa tgatgagcac tttcaaagtt ctgctatgtg gcgcggtgtt atcccgtatt
     2461 gatgctgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     2521 tattcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgtagt
     2581 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gattggagga
     2641 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     2701 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     2761 gcaatggcaa caactctccg caaactatta actggcgaac tacttactct tgcttcacgc
     2821 cagcaactca ttgactggat ggaggcggac aaagttgcag gaccacttct gcgctcggca
     2881 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     2941 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     3001 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     3061 attaagcatt ggtaactgtc agacgaaggg aaataaatag tagcccgcct gatgtgcggg
     3121 cttttttttg gtggtggtta ccagtgtcgc ttagctaccg taggtcctct taggcggtcc
     3181 gattagcgac atagtcatca at
//
