LOCUS       sequence-181901-        3143 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Module B, Promoter: TaU3, 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 3143)
  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 3143)
  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_B2517
FEATURES             Location/Qualifiers
     source          1..3143
                     /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(1278..1583)
                     /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(1380..1399)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     misc_RNA        1910..1985
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2051..2155
                     /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 ggcggcaggg agagttttaa cattgactag
      901 cgtgctgata atttgtgaga aataataatt gacaagtaga tactgacatt tgagaagagc
      961 ttctgaactg ttattagtaa caaaaatgga aagctgatgc acggaaaaag gaaagaaaaa
     1021 gccatacttt tttttaggta ggaaaagaaa aagccatacg agactgatgt ctctcagatg
     1081 ggccgggatc tgtctatcta gcaggcagca gccctaccaa cctcacgggc cagcaattac
     1141 gagtccttct aaaacgtccc gccgagggcg cgtggccgtg ctgtgcagca gcacgtctaa
     1201 cattagtccc acctcgccag tttacaggga gcagaaccag cttataagcg gaggcgcggc
     1261 accaagaagc ggagacgtta tattccccag aacatcaggt taatggcgtt tttgatgtca
     1321 ttttcgcggt ggctgagatc agccacttct tccccgataa cggaaaccgg cacactggcc
     1381 atatcggtgg tcatcatgcg ccagctttca tccccgatat gcaccaccgg gtaaagttca
     1441 cgggagactt tatctgacag cagacgtgca ctggccaggg ggatcaccat ccgtcgcccg
     1501 ggcgtgtcaa taatatcact ctgtacatcc acaaacagac gataacggct ctctctttta
     1561 taggtgtaaa ccttaaactg catttcacca gcccctgttc tcgtcagcaa aagagccgtt
     1621 catttcaata aaccgggcga cctcagccat cccttcctga ttttccgctt tccagcgttc
     1681 ggcacgcaga cgacgggctt cattctgcat ggttgtgctt accagaccgg agatattgac
     1741 atcatatatg ccttgagcaa ctgatagctg tcgctgtcaa ctgtcactgt aatacgctgc
     1801 ttcatagcat acctcctttt gacatacttc gggtatacat atcagtatat attcttatac
     1861 cgcaaaaatc agcgcgcaaa tacgcatact gttatctggc ttcgtctcag ttttagagct
     1921 agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc
     1981 ggtgcttttt ttccggtgac gcaggtgatg aaaagcctta gggtcaggtg gcacttttcg
     2041 gggaaatgtg cgatgaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2101 gctaatgaga caataaccct gataaatgct tcaattatat tgaaaaagga agagtatgag
     2161 tatccaacat ttccgtgtcg cccttattcc cttttttgcg gccttctgcc ttcctgtttt
     2221 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcccgagt
     2281 gggttacatt gaactggatc tcaacagcgg caaaatctta gagagttttc gccccgaaga
     2341 acgcttccca atgatgagca ctttcaaagt tctgctatgt ggcgcggtgt tatcccgtat
     2401 tgatgctggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     2461 gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgtag
     2521 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgattggagg
     2581 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     2641 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     2701 agcaatggca acaactctcc gcaaactatt aactggcgaa ctacttactc ttgcttcacg
     2761 ccagcaactc attgactgga tggaggcgga caaagttgca ggaccacttc tgcgctcggc
     2821 acttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     2881 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     2941 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     3001 gattaagcat tggtaactgt cagacgaagg gaaataaata gtagcccgcc tgatgtgcgg
     3061 gctttttttt ggtggtggtt accagtgtcg cttagctacc gtaggtcctc ttaggcggtc
     3121 cgattagcga catagtcatc aat
//
