LOCUS       sequence-348309-        3781 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty backbone for Golden Gate cloning into Cermak, et al., 2017 
            Toolkit.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3781)
  AUTHORS   Cody JP, Maher MF, Nasti RA, Starker CG, Chamness JC, Voytas DF
  TITLE     Direct delivery and fast-treated Agrobacterium co-culture 
            (Fast-TrACC) plant transformation methods for Nicotiana benthamiana.
  JOURNAL   Nat Protoc. 2022 Oct 17. pii: 10.1038/s41596-022-00749-9. doi: 
            10.1038/s41596-022-00749-9.
  PUBMED    36253612
REFERENCE   2  (bases 1 to 3781)
  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: "Nat Protoc.
            2022 Oct 17. pii: 10.1038/s41596-022-00749-9. doi: 
            10.1038/s41596-022-00749-9."
COMMENT     Sequence Label: pMOD_D4800EC
FEATURES             Location/Qualifiers
     source          1..3781
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    45..169
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(210..515)
                     /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
                     VPRELYPVVHIGDESWRMMTTDMASVPASVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(312..331)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(835..1515)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     promoter        complement(join(1569..1575,1576..1593,1594..1599))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(1624..1748)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1834..1938
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3006..3594
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3495..3514
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3748..3765
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cacctgcaac gtcccggcgc gccctatcga gacggatatc ggtgaccact ttgtacaaga
       61 aagctgaacg agaaacgtaa aatgatataa atatcaatat attaaattag attttgcata
      121 aaaaacagac tacataatac tgtaaaacac aacatatcca gtcactatgg tcgacctgca
      181 gactggctgt gtataaggga gcctgacatt tatattcccc agaacatcag gttaatggcg
      241 tttttgatgt cattttcgcg gtggctgaga tcagccactt cttccccgat aacggaggcc
      301 ggcacactgg ccatatcggt ggtcatcatg cgccagcttt catccccgat atgcaccacc
      361 gggtaaagtt cacgggggac tttatctgac agcagacgtg cactggccag ggggatcacc
      421 atccgtcgcc cgggcgtgtc aataatatca ctctgtacat ccacaaacag acgataacgg
      481 ctctctcttt tataggtgta aaccttaaac tgcatttcac cagcccctgt tctcgtcagc
      541 aaaagagccg ttcatttcaa taaaccgggc gacctcagcc atcccttcct gattttccgc
      601 tttccagcgt tcggcacgca gacgacgggc ttcattctgc atggttgtgc ttaccagacc
      661 ggagatattg acatcatata tgccttgagc aactgatagc tgtcgctgtc aactgtcact
      721 gtaatacgct gcttcatagc atacctcttt ttgacatact tctggtatac atatcagtat
      781 atattcttat accgcaaaaa tcagcgcgca aatacgcata ctgttatctg gcttttagta
      841 agccggatcc tctagattac gccccgcctg ccactcatcg cagtactgtt gtaattcatt
      901 aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa tcgccagcgg
      961 catcagcacc ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa
     1021 gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg gattggctga
     1081 gacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt caccgtaaca
     1141 cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca
     1201 gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt gaacactatc
     1261 ccatatcacc agctcaccgt ctttcattgc catacggaat tccggatgag cattcatcag
     1321 gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct ttacggtctt
     1381 taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacattgag cgactgactg
     1441 aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg tatatccagt
     1501 gatttttttc tccattttag cttccttagc tcctgaaaat ctcgacggat cctaactcaa
     1561 aatccacaca ttatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgcggc
     1621 cgccatagtg actggatatg ttgtgtttta cagtattatg tagtctgttt tttatgcaaa
     1681 atctaattta atatattgat atttatatca ttttacgttt ctcgttcagc ttttttgtac
     1741 aaacttgtca ccgatatccg tctccgttag agctcagtga ctagcaggtg atgaaaagcc
     1801 ttagggtcag gtggcacttt tcggggaaat gtgcgatgaa cccctatttg tttatttttc
     1861 taaatacatt caaatatgta tccgctaatg agacaataac cctgataaat gcttcaatta
     1921 tattgaaaaa ggaagagtat gagtatccaa catttccgtg tcgcccttat tccctttttt
     1981 gcggccttct gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     2041 gaagatcagt tgggtgcccg agtgggttac attgaactgg atctcaacag cggcaaaatc
     2101 ttagagagtt ttcgccccga agaacgcttc ccaatgatga gcactttcaa agttctgcta
     2161 tgtggcgcgg tgttatcccg tattgatgct gggcaagagc aactcggtcg ccgcatacac
     2221 tattctcaga atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc
     2281 atgacagtaa gagaattatg tagtgctgcc ataaccatga gtgataacac tgcggccaac
     2341 ttacttctga caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     2401 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     2461 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaactc tccgcaaact attaactggc
     2521 gaactactta ctcttgcttc acgccagcaa ctcattgact ggatggaggc ggacaaagtt
     2581 gcaggaccac ttctgcgctc ggcacttccg gctggctggt ttattgctga taaatctgga
     2641 gccggtgagc gtggctctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     2701 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     2761 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacga agggaaataa
     2821 atagtagccc gcctgatgtg cgggcttttt tttggtggtg gttaccagtg tcgcttagct
     2881 accgtaggtc ctcttaggcg gtccgattag cgacatagtc atcaatgcct tgacggcaaa
     2941 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     3001 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3061 ctaccagcgg tggtttgttt gccggatcaa gagctactaa ctctttttcc gaaggtaact
     3121 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     3181 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3241 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     3301 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgct cacagcccag cttggagcga
     3361 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     3421 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     3481 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3541 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3601 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     3661 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     3721 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     3781 c
//
