LOCUS       sequence-181880-        3500 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Module B, Promoter: CmYLCV, Gene: SapI ccdb cassette for cloning 
            multiple gRNA spacers with ribozyme spacers , Terminator: 35S.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3500)
  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 3500)
  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_B2403
FEATURES             Location/Qualifiers
     source          1..3500
                     /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"
     misc_RNA        1374..1449
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             1825..2130
                     /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     2009..2028
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    2158..2332
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        2409..2513
                     /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 aagcgggaga gcgcccacct gccaggggac ggcgcgccga agagctggca gacatactgt
      901 cccacaaatg aagatggaat ctgtaaaaga aaacgcgtga aataatgcgt ctgacaaagg
      961 ttaggtcggc tgcctttaat caataccaaa gtggtcccta ccacgatgga aaaactgtgc
     1021 agtcggtttg gctttttctg acgaacaaat aagattcgtg gccgacaggt gggggtccac
     1081 catgtgaagg catcttcaga ctccaataat ggagcaatga cgtaagggct tacgaaataa
     1141 gtaagggtag tttgggaaat gtccactcac ccgtcagtct ataaatactt agcccctccc
     1201 tcattgttaa gggagcaaaa tctcagagag atagtcctag agagagaaag agagcaagta
     1261 gcctagaagt agtcaaggcg gcgaagtatt caggcacgtg gccaggaaga agaaaagcca
     1321 agacgacgaa aacaggtaag agctaagctt cctgcagggc ttttgtcacc ggagttttag
     1381 agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg
     1441 agtcggtgct tttgtcaccg gatgtgtttt ccggtctgat gagtccgtga ggacgaaaca
     1501 aaagcaagcc agataacagt atgcgtattt gcgcgctgat ttttgcggta taagaatata
     1561 tactgatatg tatacccgaa gtatgtcaaa aggaggtatg ctatgaagca gcgtattaca
     1621 gtgacagttg acagcgacag ctatcagttg ctcaaggcat atatgatgtc aatatctccg
     1681 gtctggtaag cacaaccatg cagaatgaag cccgtcgtct gcgtgccgaa cgctggaaag
     1741 cggaaaatca ggaagggatg gctgaggtcg cccggtttat tgaaatgaac ggctcttttg
     1801 ctgacgagaa caggggctgg tgaaatgcag tttaaggttt acacctataa aagagagagc
     1861 cgttatcgtc tgtttgtgga tgtacagagt gatattattg acacgcccgg gcgacggatg
     1921 gtgatccccc tggccagtgc acgtctgctg tcagataaag tctcccgtga actttacccg
     1981 gtggtgcata tcggggatga aagctggcgc atgatgacca ccgatatggc cagtgtgccg
     2041 gtttccgtta tcggggaaga agtggctgat ctcagccacc gcgaaaatga catcaaaaac
     2101 gccattaacc tgatgttctg gggaatataa gctcttcagt cgatcgacaa gctcgagttt
     2161 ctccataata atgtgtgagt agttcccaga taagggaatt agggttccta tagggtttcg
     2221 ctcatgtgtt gagcatataa gaaaccctta gtatgtattt gtatttgtaa aatacttcta
     2281 tcaataaaat ttctaattcc taaaaccaaa atccagtact aaaatccaga tcccccgaat
     2341 tagagctcta ccggtgacgc aggtgatgaa aagccttagg gtcaggtggc acttttcggg
     2401 gaaatgtgcg atgaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     2461 taatgagaca ataaccctga taaatgcttc aattatattg aaaaaggaag agtatgagta
     2521 tccaacattt ccgtgtcgcc cttattccct tttttgcggc cttctgcctt cctgtttttg
     2581 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcccgagtgg
     2641 gttacattga actggatctc aacagcggca aaatcttaga gagttttcgc cccgaagaac
     2701 gcttcccaat gatgagcact ttcaaagttc tgctatgtgg cgcggtgtta tcccgtattg
     2761 atgctgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     2821 attcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgtagtg
     2881 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac
     2941 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     3001 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     3061 caatggcaac aactctccgc aaactattaa ctggcgaact acttactctt gcttcacgcc
     3121 agcaactcat tgactggatg gaggcggaca aagttgcagg accacttctg cgctcggcac
     3181 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggc tctcgcggta
     3241 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     3301 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     3361 ttaagcattg gtaactgtca gacgaaggga aataaatagt agcccgcctg atgtgcgggc
     3421 ttttttttgg tggtggttac agtgtcgctt agctaccgta ggtcctctta ggcggtccga
     3481 ttagcgacat agtcatcaat
//
