LOCUS       sequence-182048-        3139 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Module C, Promoter: At7SL, 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 3139)
  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 3139)
  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_C2516
FEATURES             Location/Qualifiers
     source          1..3139
                     /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(1203..1508)
                     /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(1305..1324)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     misc_RNA        1835..1910
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2047..2151
                     /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 cgccatggat gttgttgtta ccagaaagta
      901 aataaatgtt caatctctga tgttctcaag taagtgagtt ttattgggaa taatattaac
      961 ttatgttctt cttgcatttg atttctttgc cgctctcttc ttctatctta aatctgtgta
     1021 tactatttca ctattgggct ttttattagt ctataatggg actcaaaata aggctttggc
     1081 ccacatcaaa aagataagtc acaaatcaaa actaaattca gagtcttttc tcccacatcg
     1141 gtcactgtac tcattttgtg tttgtttata tattacacga accgatcttt ggtacggaga
     1201 cgttatattc cccagaacat caggttaatg gcgtttttga tgtcattttc gcggtggctg
     1261 agatcagcca cttcttcccc gataacggaa accggcacac tggccatatc ggtggtcatc
     1321 atgcgccagc tttcatcccc gatatgcacc accgggtaaa gttcacggga gactttatct
     1381 gacagcagac gtgcactggc cagggggatc accatccgtc gcccgggcgt gtcaataata
     1441 tcactctgta catccacaaa cagacgataa cggctctctc ttttataggt gtaaacctta
     1501 aactgcattt caccagcccc tgttctcgtc agcaaaagag ccgttcattt caataaaccg
     1561 ggcgacctca gccatccctt cctgattttc cgctttccag cgttcggcac gcagacgacg
     1621 ggcttcattc tgcatggttg tgcttaccag accggagata ttgacatcat atatgccttg
     1681 agcaactgat agctgtcgct gtcaactgtc actgtaatac gctgcttcat agcatacctc
     1741 cttttgacat acttcgggta tacatatcag tatatattct tataccgcaa aaatcagcgc
     1801 gcaaatacgc atactgttat ctggcttcgt ctctgtttta gagctagaaa tagcaagtta
     1861 aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc tttttttcgc
     1921 gcgtagtcct cggtaatatc gcagaacaaa agtacctgat atcgagtgta cttcaagtca
     1981 caccggcgag tgactagcag gtgatgaaaa gccttagggt caggtggcac ttttcgggga
     2041 aatgtgcgat gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgcta
     2101 atgagacaat aaccctgata aatgcttcaa ttatattgaa aaaggaagag tatgagtatc
     2161 caacatttcc gtgtcgccct tattcccttt tttgcggcct tctgccttcc tgtttttgct
     2221 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc ccgagtgggt
     2281 tacattgaac tggatctcaa cagcggcaaa atcttagaga gttttcgccc cgaagaacgc
     2341 ttcccaatga tgagcacttt caaagttctg ctatgtggcg cggtgttatc ccgtattgat
     2401 gctgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtat
     2461 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgtagtgct
     2521 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat tggaggaccg
     2581 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     2641 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     2701 atggcaacaa ctctccgcaa actattaact ggcgaactac ttactcttgc ttcacgccag
     2761 caactcattg actggatgga ggcggacaaa gttgcaggac cacttctgcg ctcggcactt
     2821 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     2881 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     2941 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     3001 aagcattggt aactgtcaga cgaagggaaa taaatagtag cccgcctgat gtgcgggctt
     3061 ttttttggtg gtggttacca gtgtcgctta gctaccgtag gtcctcttag gcggtccgat
     3121 tagcgacata gtcatcaat
//
