LOCUS       sequence-181874-        3584 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Module B, Promoter: CmYLCV, Gene: SapI ccdb cassette for cloning 
            multiple gRNA spacers with tRNA spacers , Terminator: 35S.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3584)
  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 3584)
  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_B2303
FEATURES             Location/Qualifiers
     source          1..3584
                     /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        1436..1511
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             1908..2213
                     /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     2092..2111
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    2241..2415
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        2492..2596
                     /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 cctgcaggaa caaagcacca gtggtctagt
     1381 ggtagaatag taccctgcca cggtacagac ccgggttcga ttcccggctg gtgcagtttt
     1441 agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac
     1501 cgagtcggtg caacaaagca ccagtggtct agtggtagaa tagtaccctg ccacggtaca
     1561 gacccgggtt cgattcccgg ctggtgcaaa gccagataac agtatgcgta tttgcgcgct
     1621 gatttttgcg gtataagaat atatactgat atgtataccc gaagtatgtc aaaaggaggt
     1681 atgctatgaa gcagcgtatt acagtgacag ttgacagcga cagctatcag ttgctcaagg
     1741 catatatgat gtcaatatct ccggtctggt aagcacaacc atgcagaatg aagcccgtcg
     1801 tctgcgtgcc gaacgctgga aagcggaaaa tcaggaaggg atggctgagg tcgcccggtt
     1861 tattgaaatg aacggctctt ttgctgacga gaacaggggc tggtgaaatg cagtttaagg
     1921 tttacaccta taaaagagag agccgttatc gtctgtttgt ggatgtacag agtgatatta
     1981 ttgacacgcc cgggcgacgg atggtgatcc ccctggccag tgcacgtctg ctgtcagata
     2041 aagtctcccg tgaactttac ccggtggtgc atatcgggga tgaaagctgg cgcatgatga
     2101 ccaccgatat ggccagtgtg ccggtttccg ttatcgggga agaagtggct gatctcagcc
     2161 accgcgaaaa tgacatcaaa aacgccatta acctgatgtt ctggggaata taagctcttc
     2221 agtcgatcga caagctcgag tttctccata ataatgtgtg agtagttccc agataaggga
     2281 attagggttc ctatagggtt tcgctcatgt gttgagcata taagaaaccc ttagtatgta
     2341 tttgtatttg taaaatactt ctatcaataa aatttctaat tcctaaaacc aaaatccagt
     2401 actaaaatcc agatcccccg aattagagct ctaccggtga cgcaggtgat gaaaagcctt
     2461 agggtcaggt ggcacttttc ggggaaatgt gcgatgaacc cctatttgtt tatttttcta
     2521 aatacattca aatatgtatc cgctaatgag acaataaccc tgataaatgc ttcaattata
     2581 ttgaaaaagg aagagtatga gtatccaaca tttccgtgtc gcccttattc ccttttttgc
     2641 ggccttctgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     2701 agatcagttg ggtgcccgag tgggttacat tgaactggat ctcaacagcg gcaaaatctt
     2761 agagagtttt cgccccgaag aacgcttccc aatgatgagc actttcaaag ttctgctatg
     2821 tggcgcggtg ttatcccgta ttgatgctgg gcaagagcaa ctcggtcgcc gcatacacta
     2881 ttctcagaat gacttggttg agtattcacc agtcacagaa aagcatctta cggatggcat
     2941 gacagtaaga gaattatgta gtgctgccat aaccatgagt gataacactg cggccaactt
     3001 acttctgaca acgattggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     3061 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     3121 gcgtgacacc acgatgcctg tagcaatggc aacaactctc cgcaaactat taactggcga
     3181 actacttact cttgcttcac gccagcaact cattgactgg atggaggcgg acaaagttgc
     3241 aggaccactt ctgcgctcgg cacttccggc tggctggttt attgctgata aatctggagc
     3301 cggtgagcgt ggctctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     3361 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     3421 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagacgaag ggaaataaat
     3481 agtagcccgc ctgatgtgcg ggcttttttt tggtggtggt taccagtgtc gcttagctac
     3541 cgtaggtcct cttaggcggt ccgattagcg acatagtcat caat
//
