LOCUS       sequence-348311-        3782 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 3782)
  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 3782)
  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_A4800EC
FEATURES             Location/Qualifiers
     source          1..3782
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      95..683
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     584..603
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     837..854
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    915..1039
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1080..1385)
                     /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(1182..1201)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(1705..2385)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     promoter        complement(join(2439..2445,2446..2463,2464..2469))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(2494..2618)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        2705..2809
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacatagtca tcaatgcctt gacggcaaaa tcccttaacg tgagttttcg ttccactgag
       61 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
      121 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
      181 agctactaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
      241 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
      301 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
      361 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
      421 gttcgtgctc acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
      481 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
      541 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
      601 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
      661 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
      721 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
      781 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
      841 agtcagtgag cgaggaagcg gaagagcgcc cacctgctca tgatcggcgc gccctatcga
      901 gacggatatc ggtgaccact ttgtacaaga aagctgaacg agaaacgtaa aatgatataa
      961 atatcaatat attaaattag attttgcata aaaaacagac tacataatac tgtaaaacac
     1021 aacatatcca gtcactatgg tcgacctgca gactggctgt gtataaggga gcctgacatt
     1081 tatattcccc agaacatcag gttaatggcg tttttgatgt cattttcgcg gtggctgaga
     1141 tcagccactt cttccccgat aacggaggcc ggcacactgg ccatatcggt ggtcatcatg
     1201 cgccagcttt catccccgat atgcaccacc gggtaaagtt cacgggggac tttatctgac
     1261 agcagacgtg cactggccag ggggatcacc atccgtcgcc cgggcgtgtc aataatatca
     1321 ctctgtacat ccacaaacag acgataacgg ctctctcttt tataggtgta aaccttaaac
     1381 tgcatttcac cagcccctgt tctcgtcagc aaaagagccg ttcatttcaa taaaccgggc
     1441 gacctcagcc atcccttcct gattttccgc tttccagcgt tcggcacgca gacgacgggc
     1501 ttcattctgc atggttgtgc ttaccagacc ggagatattg acatcatata tgccttgagc
     1561 aactgatagc tgtcgctgtc aactgtcact gtaatacgct gcttcatagc atacctcttt
     1621 ttgacatact tctggtatac atatcagtat atattcttat accgcaaaaa tcagcgcgca
     1681 aatacgcata ctgttatctg gcttttagta agccggatcc tctagattac gccccgcctg
     1741 ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca
     1801 aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata
     1861 tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa
     1921 actggtgaaa ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt
     1981 agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa
     2041 ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg
     2101 gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc
     2161 catacggaat tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata
     2221 aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt
     2281 ctggttatag gtacattgag cgactgactg aaatgcctca aaatgttctt tacgatgcca
     2341 ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc
     2401 tcctgaaaat ctcgacggat cctaactcaa aatccacaca ttatacgagc cggaagcata
     2461 aagtgtaaag cctggggtgc ctaatgcggc cgccatagtg actggatatg ttgtgtttta
     2521 cagtattatg tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca
     2581 ttttacgttt ctcgttcagc ttttttgtac aaacttgtca cgatatccgt ctccgttaga
     2641 gctcggacgt gggcaggtgt gatgaaaagc cttagggtca ggtggcactt ttcggggaaa
     2701 tgtgcgatga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctaat
     2761 gagacaataa ccctgataaa tgcttcaatt atattgaaaa aggaagagta tgagtatcca
     2821 acatttccgt gtcgccctta ttcccttttt tgcggccttc tgccttcctg tttttgctca
     2881 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgccc gagtgggtta
     2941 cattgaactg gatctcaaca gcggcaaaat cttagagagt tttcgccccg aagaacgctt
     3001 cccaatgatg agcactttca aagttctgct atgtggcgcg gtgttatccc gtattgatgc
     3061 tgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtattc
     3121 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gtagtgctgc
     3181 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg gaggaccgaa
     3241 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     3301 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     3361 ggcaacaact ctccgcaaac tattaactgg cgaactactt actcttgctt cacgccagca
     3421 actcattgac tggatggagg cggacaaagt tgcaggacca cttctgcgct cggcacttcc
     3481 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtggctctc gcggtatcat
     3541 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     3601 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     3661 gcattggtaa ctgtcagacg aagggaaata aatagtagcc cgcctgatgt gcgggctttt
     3721 ttttggtggt ggttaccagt gtcgcttagc taccgtaggt cctcttaggc ggtccgatta
     3781 gc
//
