LOCUS       sequence-181899-        3195 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Module B, Promoter: AtU6, Gene: Esp3I ccdb cassette for single gRNA 
            spacer cloning, Terminator: Pol III.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3195)
  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 3195)
  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_B2515
FEATURES             Location/Qualifiers
     source          1..3195
                     /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(1329..1634)
                     /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(1431..1450)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     misc_RNA        1961..2036
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2103..2207
                     /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 gccaggggac ccatggcttc gttgaacaac ggaaactcga
      901 cttgccttcc gcacaataca tcatttcttc ttagcttttt ttcttcttct tcgttcatac
      961 agtttttttt tgtttatcag cttacatttt cttgaaccgt agctttcgtt ttcttctttt
     1021 taactttcca ttcggagttt ttgtatcttg tttcatagtt tgtcccagga ttagaatgat
     1081 taggcatcga accttcaaga atttgattga ataaaacatc ttcattctta agatatgaag
     1141 ataatcttca aaaggcccct gggaatctga aagaagagaa gcaggcccat ttatatggga
     1201 aagaacaata gtatttctta tataggccca tttaagttga aaacaatctt caaaagtccc
     1261 acatcgctta gataagaaaa cgaagctgag tttatataca gctagagtcg aagtagtgat
     1321 tggagacgtt atattcccca gaacatcagg ttaatggcgt ttttgatgtc attttcgcgg
     1381 tggctgagat cagccacttc ttccccgata acggaaaccg gcacactggc catatcggtg
     1441 gtcatcatgc gccagctttc atccccgata tgcaccaccg ggtaaagttc acgggagact
     1501 ttatctgaca gcagacgtgc actggccagg gggatcacca tccgtcgccc gggcgtgtca
     1561 ataatatcac tctgtacatc cacaaacaga cgataacggc tctctctttt ataggtgtaa
     1621 accttaaact gcatttcacc agcccctgtt ctcgtcagca aaagagccgt tcatttcaat
     1681 aaaccgggcg acctcagcca tcccttcctg attttccgct ttccagcgtt cggcacgcag
     1741 acgacgggct tcattctgca tggttgtgct taccagaccg gagatattga catcatatat
     1801 gccttgagca actgatagct gtcgctgtca actgtcactg taatacgctg cttcatagca
     1861 tacctccttt tgacatactt cgggtataca tatcagtata tattcttata ccgcaaaaat
     1921 cagcgcgcaa atacgcatac tgttatctgg cttcgtctct gttttagagc tagaaatagc
     1981 aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt
     2041 tttcccggtg acgcaggtga tgaaaagcct tagggtcagg tggcactttt cggggaaatg
     2101 tgcgatgaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctaatga
     2161 gacaataacc ctgataaatg cttcaattat attgaaaaag gaagagtatg agtatccaac
     2221 atttccgtgt cgcccttatt cccttttttg cggccttctg ccttcctgtt tttgctcacc
     2281 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcccga gtgggttaca
     2341 ttgaactgga tctcaacagc ggcaaaatct tagagagttt tcgccccgaa gaacgcttcc
     2401 caatgatgag cactttcaaa gttctgctat gtggcgcggt gttatcccgt attgatgctg
     2461 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtattcac
     2521 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgt agtgctgcca
     2581 taaccatgag tgataacact gcggccaact tacttctgac aacgattgga ggaccgaagg
     2641 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     2701 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     2761 caacaactct ccgcaaacta ttaactggcg aactacttac tcttgcttca cgccagcaac
     2821 tcattgactg gatggaggcg gacaaagttg caggaccact tctgcgctcg gcacttccgg
     2881 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     2941 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     3001 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     3061 attggtaact gtcagacgaa gggaaataaa tagtagcccg cctgatgtgc gggctttttt
     3121 ttggtggtgg ttaccagtgt cgcttagcta ccgtaggtcc tcttaggcgg tccgattagc
     3181 gacatagtca tcaat
//
