LOCUS       sequence-181905-        2996 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Module B, Promoter: OsU6, 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 2996)
  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 2996)
  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_B2520
FEATURES             Location/Qualifiers
     source          1..2996
                     /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(1130..1435)
                     /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(1232..1251)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     misc_RNA        1762..1837
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        1904..2008
                     /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 ccatgggcgg gatcatgaac caacggcctg
      901 gctgtatttg gtggttgtgt agggagatgg ggagaagaaa agcccgattc tcttcgctgt
      961 gatgggctgg atgcatgcgg gggagcggga ggcccaagta cgtgcacggt gagcggccca
     1021 cagggcgagt gtgagcgcga gaggcgggag gaacagttta gtaccacatt gcccagctaa
     1081 ctcgaacgcg accaacttat aaacccgcgc gctgtcgctt gtggagacgt tatattcccc
     1141 agaacatcag gttaatggcg tttttgatgt cattttcgcg gtggctgaga tcagccactt
     1201 cttccccgat aacggaaacc ggcacactgg ccatatcggt ggtcatcatg cgccagcttt
     1261 catccccgat atgcaccacc gggtaaagtt cacgggagac tttatctgac agcagacgtg
     1321 cactggccag ggggatcacc atccgtcgcc cgggcgtgtc aataatatca ctctgtacat
     1381 ccacaaacag acgataacgg ctctctcttt tataggtgta aaccttaaac tgcatttcac
     1441 cagcccctgt tctcgtcagc aaaagagccg ttcatttcaa taaaccgggc gacctcagcc
     1501 atcccttcct gattttccgc tttccagcgt tcggcacgca gacgacgggc ttcattctgc
     1561 atggttgtgc ttaccagacc ggagatattg acatcatata tgccttgagc aactgatagc
     1621 tgtcgctgtc aactgtcact gtaatacgct gcttcatagc atacctcctt ttgacatact
     1681 tcgggtatac atatcagtat atattcttat accgcaaaaa tcagcgcgca aatacgcata
     1741 ctgttatctg gcttcgtctc tgttttagag ctagaaatag caagttaaaa taaggctagt
     1801 ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt ttttcccggt gacgcaggtg
     1861 atgaaaagcc ttagggtcag gtggcacttt tcggggaaat gtgcgatgaa cccctatttg
     1921 tttatttttc taaatacatt caaatatgta tccgctaatg agacaataac cctgataaat
     1981 gcttcaatta tattgaaaaa ggaagagtat gagtatccaa catttccgtg tcgcccttat
     2041 tccctttttt gcggccttct gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     2101 aaaagatgct gaagatcagt tgggtgcccg agtgggttac attgaactgg atctcaacag
     2161 cggcaaaatc ttagagagtt ttcgccccga agaacgcttc ccaatgatga gcactttcaa
     2221 agttctgcta tgtggcgcgg tgttatcccg tattgatgct gggcaagagc aactcggtcg
     2281 ccgcatacac tattctcaga atgacttggt tgagtattca ccagtcacag aaaagcatct
     2341 tacggatggc atgacagtaa gagaattatg tagtgctgcc ataaccatga gtgataacac
     2401 tgcggccaac ttacttctga caacgattgg aggaccgaag gagctaaccg cttttttgca
     2461 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     2521 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaactc tccgcaaact
     2581 attaactggc gaactactta ctcttgcttc acgccagcaa ctcattgact ggatggaggc
     2641 ggacaaagtt gcaggaccac ttctgcgctc ggcacttccg gctggctggt ttattgctga
     2701 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     2761 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     2821 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacga
     2881 agggaaataa atagtagccc gcctgatgtg cgggcttttt tttggtggtg gttaccagtg
     2941 tcgcttagct accgtaggtc ctcttaggcg gtccgattag cgacatagtc atcaat
//
