LOCUS       sequence-182054-        3059 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Module C, Promoter: OsU6, 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 3059)
  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 3059)
  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_C2520
FEATURES             Location/Qualifiers
     source          1..3059
                     /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"
     CDS             complement(1123..1428)
                     /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(1225..1244)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     misc_RNA        1755..1830
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        1967..2071
                     /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 gcaacgccgg cgggatcatg aaccaacggc ctggctgtat
      901 ttggtggttg tgtagggaga tggggagaag aaaagcccga ttctcttcgc tgtgatgggc
      961 tggatgcatg cgggggagcg ggaggcccaa gtacgtgcac ggtgagcggc ccacagggcg
     1021 agtgtgagcg cgagaggcgg gaggaacagt ttagtaccac attgcccagc taactcgaac
     1081 gcgaccaact tataaacccg cgcgctgtcg cttgtggaga cgttatattc cccagaacat
     1141 caggttaatg gcgtttttga tgtcattttc gcggtggctg agatcagcca cttcttcccc
     1201 gataacggaa accggcacac tggccatatc ggtggtcatc atgcgccagc tttcatcccc
     1261 gatatgcacc accgggtaaa gttcacggga gactttatct gacagcagac gtgcactggc
     1321 cagggggatc accatccgtc gcccgggcgt gtcaataata tcactctgta catccacaaa
     1381 cagacgataa cggctctctc ttttataggt gtaaacctta aactgcattt caccagcccc
     1441 tgttctcgtc agcaaaagag ccgttcattt caataaaccg ggcgacctca gccatccctt
     1501 cctgattttc cgctttccag cgttcggcac gcagacgacg ggcttcattc tgcatggttg
     1561 tgcttaccag accggagata ttgacatcat atatgccttg agcaactgat agctgtcgct
     1621 gtcaactgtc actgtaatac gctgcttcat agcatacctc cttttgacat acttcgggta
     1681 tacatatcag tatatattct tataccgcaa aaatcagcgc gcaaatacgc atactgttat
     1741 ctggcttcgt ctctgtttta gagctagaaa tagcaagtta aaataaggct agtccgttat
     1801 caacttgaaa aagtggcacc gagtcggtgc tttttttcgc gcgtagtcct cggtaatatc
     1861 gcagaacaaa agtacctgat atcgagtgta cttcaagtca caccggcgag tgactagcag
     1921 gtgatgaaaa gccttagggt caggtggcac ttttcgggga aatgtgcgat gaacccctat
     1981 ttgtttattt ttctaaatac attcaaatat gtatccgcta atgagacaat aaccctgata
     2041 aatgcttcaa ttatattgaa aaaggaagag tatgagtatc caacatttcc gtgtcgccct
     2101 tattcccttt tttgcggcct tctgccttcc tgtttttgct cacccagaaa cgctggtgaa
     2161 agtaaaagat gctgaagatc agttgggtgc ccgagtgggt tacattgaac tggatctcaa
     2221 cagcggcaaa atcttagaga gttttcgccc cgaagaacgc ttcccaatga tgagcacttt
     2281 caaagttctg ctatgtggcg cggtgttatc ccgtattgat gctgggcaag agcaactcgg
     2341 tcgccgcata cactattctc agaatgactt ggttgagtat tcaccagtca cagaaaagca
     2401 tcttacggat ggcatgacag taagagaatt atgtagtgct gccataacca tgagtgataa
     2461 cactgcggcc aacttacttc tgacaacgat tggaggaccg aaggagctaa ccgctttttt
     2521 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     2581 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa ctctccgcaa
     2641 actattaact ggcgaactac ttactcttgc ttcacgccag caactcattg actggatgga
     2701 ggcggacaaa gttgcaggac cacttctgcg ctcggcactt ccggctggct ggtttattgc
     2761 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     2821 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     2881 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     2941 cgaagggaaa taaatagtag cccgcctgat gtgcgggctt ttttttggtg gtggttacca
     3001 gtgtcgctta gctaccgtag gtcctcttag gcggtccgat tagcgacata gtcatcaat
//
