LOCUS       sequence-182050-        3149 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Module C, Promoter: At7SL, Gene: Esp3I ccdb cassette for single gRNA
            spacer cloning, structurally optimized gRNA scaffold, (+ 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 3149)
  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 3149)
  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_C2516b
FEATURES             Location/Qualifiers
     source          1..3149
                     /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"
     promoter        2057..2161
                     /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 ctctgtttaa gagctatgct ggaaacagca
     1861 tagcaagttt aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc
     1921 tttttttcgc gcgtagtcct cggtaatatc gcagaacaaa agtacctgat atcgagtgta
     1981 cttcaagtca caccggcgag tgactagcag gtgatgaaaa gccttagggt caggtggcac
     2041 ttttcgggga aatgtgcgat gaacccctat ttgtttattt ttctaaatac attcaaatat
     2101 gtatccgcta atgagacaat aaccctgata aatgcttcaa ttatattgaa aaaggaagag
     2161 tatgagtatc caacatttcc gtgtcgccct tattcccttt tttgcggcct tctgccttcc
     2221 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     2281 ccgagtgggt tacattgaac tggatctcaa cagcggcaaa atcttagaga gttttcgccc
     2341 cgaagaacgc ttcccaatga tgagcacttt caaagttctg ctatgtggcg cggtgttatc
     2401 ccgtattgat gctgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     2461 ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     2521 atgtagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     2581 tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     2641 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     2701 gcctgtagca atggcaacaa ctctccgcaa actattaact ggcgaactac ttactcttgc
     2761 ttcacgccag caactcattg actggatgga ggcggacaaa gttgcaggac cacttctgcg
     2821 ctcggcactt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     2881 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     2941 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     3001 ctcactgatt aagcattggt aactgtcaga cgaagggaaa taaatagtag cccgcctgat
     3061 gtgcgggctt ttttttggtg gtggttacca gtgtcgctta gctaccgtag gtcctcttag
     3121 gcggtccgat tagcgacata gtcatcaat
//
