LOCUS       sequence-181900-        3205 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Module B, Promoter: AtU6, Gene: Esp3I ccdb cassette for single gRNA 
            spacer cloning, structurally optimized gRNA scaffold, Terminator: 
            Pol III.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3205)
  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 3205)
  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_B2515b
FEATURES             Location/Qualifiers
     source          1..3205
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        29..133
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1201..1789
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1690..1709
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1943..1960
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(2450..2755)
                     /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(2552..2571)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
ORIGIN
        1 gtcaggtggc acttttcggg gaaatgtgcg atgaacccct atttgtttat ttttctaaat
       61 acattcaaat atgtatccgc taatgagaca ataaccctga taaatgcttc aattatattg
      121 aaaaaggaag agtatgagta tccaacattt ccgtgtcgcc cttattccct tttttgcggc
      181 cttctgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
      241 tcagttgggt gcccgagtgg gttacattga actggatctc aacagcggca aaatcttaga
      301 gagttttcgc cccgaagaac gcttcccaat gatgagcact ttcaaagttc tgctatgtgg
      361 cgcggtgtta tcccgtattg atgctgggca agagcaactc ggtcgccgca tacactattc
      421 tcagaatgac ttggttgagt attcaccagt cacagaaaag catcttacgg atggcatgac
      481 agtaagagaa ttatgtagtg ctgccataac catgagtgat aacactgcgg ccaacttact
      541 tctgacaacg attggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
      601 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
      661 tgacaccacg atgcctgtag caatggcaac aactctccgc aaactattaa ctggcgaact
      721 acttactctt gcttcacgcc agcaactcat tgactggatg gaggcggaca aagttgcagg
      781 accacttctg cgctcggcac ttccggctgg ctggtttatt gctgataaat ctggagccgg
      841 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
      901 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
      961 tgagataggt gcctcactga ttaagcattg gtaactgtca gacgaaggga aataaatagt
     1021 agcccgcctg atgtgcgggc ttttttttgg tggtggttac cagtgtcgct tagctaccgt
     1081 aggtcctctt aggcggtccg attagcgaca tagtcatcaa tgccttgacg gcaaaatccc
     1141 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     1201 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     1261 agcggtggtt tgtttgccgg atcaagagct actaactctt tttccgaagg taactggctt
     1321 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     1381 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     1441 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     1501 ggcgcagcgg tcgggctgaa cggggggttc gtgctcacag cccagcttgg agcgaacgac
     1561 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     1621 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     1681 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     1741 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     1801 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     1861 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     1921 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccacc
     1981 tgccagggga cccatggctt cgttgaacaa cggaaactcg acttgccttc cgcacaatac
     2041 atcatttctt cttagctttt tttcttcttc ttcgttcata cagttttttt ttgtttatca
     2101 gcttacattt tcttgaaccg tagctttcgt tttcttcttt ttaactttcc attcggagtt
     2161 tttgtatctt gtttcatagt ttgtcccagg attagaatga ttaggcatcg aaccttcaag
     2221 aatttgattg aataaaacat cttcattctt aagatatgaa gataatcttc aaaaggcccc
     2281 tgggaatctg aaagaagaga agcaggccca tttatatggg aaagaacaat agtatttctt
     2341 atataggccc atttaagttg aaaacaatct tcaaaagtcc cacatcgctt agataagaaa
     2401 acgaagctga gtttatatac agctagagtc gaagtagtga ttggagacgt tatattcccc
     2461 agaacatcag gttaatggcg tttttgatgt cattttcgcg gtggctgaga tcagccactt
     2521 cttccccgat aacggaaacc ggcacactgg ccatatcggt ggtcatcatg cgccagcttt
     2581 catccccgat atgcaccacc gggtaaagtt cacgggagac tttatctgac agcagacgtg
     2641 cactggccag ggggatcacc atccgtcgcc cgggcgtgtc aataatatca ctctgtacat
     2701 ccacaaacag acgataacgg ctctctcttt tataggtgta aaccttaaac tgcatttcac
     2761 cagcccctgt tctcgtcagc aaaagagccg ttcatttcaa taaaccgggc gacctcagcc
     2821 atcccttcct gattttccgc tttccagcgt tcggcacgca gacgacgggc ttcattctgc
     2881 atggttgtgc ttaccagacc ggagatattg acatcatata tgccttgagc aactgatagc
     2941 tgtcgctgtc aactgtcact gtaatacgct gcttcatagc atacctcctt ttgacatact
     3001 tcgggtatac atatcagtat atattcttat accgcaaaaa tcagcgcgca aatacgcata
     3061 ctgttatctg gcttcgtctc tgtttaagag ctatgctgga aacagcatag caagtttaaa
     3121 taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt ttttcccggt
     3181 gacgcaggtg atgaaaagcc ttagg
//
