LOCUS       sequence-182047-        3267 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Module C, Promoter: AtU6, 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 3267)
  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 3267)
  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_C2515
FEATURES             Location/Qualifiers
     source          1..3267
                     /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(1331..1636)
                     /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(1433..1452)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     misc_RNA        1963..2038
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2175..2279
                     /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 cgccatggct tcgttgaaca acggaaactc
      901 gacttgcctt ccgcacaata catcatttct tcttagcttt ttttcttctt cttcgttcat
      961 acagtttttt tttgtttatc agcttacatt ttcttgaacc gtagctttcg ttttcttctt
     1021 tttaactttc cattcggagt ttttgtatct tgtttcatag tttgtcccag gattagaatg
     1081 attaggcatc gaaccttcaa gaatttgatt gaataaaaca tcttcattct taagatatga
     1141 agataatctt caaaaggccc ctgggaatct gaaagaagag aagcaggccc atttatatgg
     1201 gaaagaacaa tagtatttct tatataggcc catttaagtt gaaaacaatc ttcaaaagtc
     1261 ccacatcgct tagataagaa aacgaagctg agtttatata cagctagagt cgaagtagtg
     1321 attggagacg ttatattccc cagaacatca ggttaatggc gtttttgatg tcattttcgc
     1381 ggtggctgag atcagccact tcttccccga taacggaaac cggcacactg gccatatcgg
     1441 tggtcatcat gcgccagctt tcatccccga tatgcaccac cgggtaaagt tcacgggaga
     1501 ctttatctga cagcagacgt gcactggcca gggggatcac catccgtcgc ccgggcgtgt
     1561 caataatatc actctgtaca tccacaaaca gacgataacg gctctctctt ttataggtgt
     1621 aaaccttaaa ctgcatttca ccagcccctg ttctcgtcag caaaagagcc gttcatttca
     1681 ataaaccggg cgacctcagc catcccttcc tgattttccg ctttccagcg ttcggcacgc
     1741 agacgacggg cttcattctg catggttgtg cttaccagac cggagatatt gacatcatat
     1801 atgccttgag caactgatag ctgtcgctgt caactgtcac tgtaatacgc tgcttcatag
     1861 catacctcct tttgacatac ttcgggtata catatcagta tatattctta taccgcaaaa
     1921 atcagcgcgc aaatacgcat actgttatct ggcttcgtct ctgttttaga gctagaaata
     1981 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
     2041 tttttcgcgc gtagtcctcg gtaatatcgc agaacaaaag tacctgatat cgagtgtact
     2101 tcaagtcaca ccggcgagtg actagcaggt gatgaaaagc cttagggtca ggtggcactt
     2161 ttcggggaaa tgtgcgatga acccctattt gtttattttt ctaaatacat tcaaatatgt
     2221 atccgctaat gagacaataa ccctgataaa tgcttcaatt atattgaaaa aggaagagta
     2281 tgagtatcca acatttccgt gtcgccctta ttcccttttt tgcggccttc tgccttcctg
     2341 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgccc
     2401 gagtgggtta cattgaactg gatctcaaca gcggcaaaat cttagagagt tttcgccccg
     2461 aagaacgctt cccaatgatg agcactttca aagttctgct atgtggcgcg gtgttatccc
     2521 gtattgatgc tgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     2581 ttgagtattc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     2641 gtagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg
     2701 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     2761 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     2821 ctgtagcaat ggcaacaact ctccgcaaac tattaactgg cgaactactt actcttgctt
     2881 cacgccagca actcattgac tggatggagg cggacaaagt tgcaggacca cttctgcgct
     2941 cggcacttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     3001 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     3061 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     3121 cactgattaa gcattggtaa ctgtcagacg aagggaaata aatagtagcc cgcctgatgt
     3181 gcgggctttt ttttggtggt ggttaccagt gtcgcttagc taccgtaggt cctcttaggc
     3241 ggtccgatta gcgacatagt catcaat
//
