LOCUS       sequence-182051-        3214 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Module C, Promoter: TaU3, 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 3214)
  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 3214)
  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_C2517
FEATURES             Location/Qualifiers
     source          1..3214
                     /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        2122..2226
                     /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 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 ttcgcgcgta gtcctcggta atatcgcaga acaaaagtac ctgatatcga
     2041 gtgtacttca agtcacaccg gcgagtgact agcaggtgat gaaaagcctt agggtcaggt
     2101 ggcacttttc ggggaaatgt gcgatgaacc cctatttgtt tatttttcta aatacattca
     2161 aatatgtatc cgctaatgag acaataaccc tgataaatgc ttcaattata ttgaaaaagg
     2221 aagagtatga gtatccaaca tttccgtgtc gcccttattc ccttttttgc ggccttctgc
     2281 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     2341 ggtgcccgag tgggttacat tgaactggat ctcaacagcg gcaaaatctt agagagtttt
     2401 cgccccgaag aacgcttccc aatgatgagc actttcaaag ttctgctatg tggcgcggtg
     2461 ttatcccgta ttgatgctgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     2521 gacttggttg agtattcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     2581 gaattatgta gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     2641 acgattggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     2701 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     2761 acgatgcctg tagcaatggc aacaactctc cgcaaactat taactggcga actacttact
     2821 cttgcttcac gccagcaact cattgactgg atggaggcgg acaaagttgc aggaccactt
     2881 ctgcgctcgg cacttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     2941 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     3001 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     3061 ggtgcctcac tgattaagca ttggtaactg tcagacgaag ggaaataaat agtagcccgc
     3121 ctgatgtgcg ggcttttttt tggtggtggt taccagtgtc gcttagctac cgtaggtcct
     3181 cttaggcggt ccgattagcg acatagtcat caat
//
