LOCUS       sequence-348312-        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_B4800EC
FEATURES             Location/Qualifiers
     source          1..3781
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    63..187
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(228..533)
                     /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(330..349)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(853..1533)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     promoter        complement(join(1587..1593,1594..1611,1612..1617))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(1642..1766)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1852..1956
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3024..3612
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3513..3532
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(3766..3781,1..2)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aggaagcggg agagcgccca cctgccaggg gacggcgcgc cctatcgaga cggatatcgg
       61 tgaccacttt gtacaagaaa gctgaacgag aaacgtaaaa tgatataaat atcaatatat
      121 taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa catatccagt
      181 cactatggtc gacctgcaga ctggctgtgt ataagggagc ctgacattta tattccccag
      241 aacatcaggt taatggcgtt tttgatgtca ttttcgcggt ggctgagatc agccacttct
      301 tccccgataa cggaggccgg cacactggcc atatcggtgg tcatcatgcg ccagctttca
      361 tccccgatat gcaccaccgg gtaaagttca cgggggactt tatctgacag cagacgtgca
      421 ctggccaggg ggatcaccat ccgtcgcccg ggcgtgtcaa taatatcact ctgtacatcc
      481 acaaacagac gataacggct ctctctttta taggtgtaaa ccttaaactg catttcacca
      541 gcccctgttc tcgtcagcaa aagagccgtt catttcaata aaccgggcga cctcagccat
      601 cccttcctga ttttccgctt tccagcgttc ggcacgcaga cgacgggctt cattctgcat
      661 ggttgtgctt accagaccgg agatattgac atcatatatg ccttgagcaa ctgatagctg
      721 tcgctgtcaa ctgtcactgt aatacgctgc ttcatagcat acctcttttt gacatacttc
      781 tggtatacat atcagtatat attcttatac cgcaaaaatc agcgcgcaaa tacgcatact
      841 gttatctggc ttttagtaag ccggatcctc tagattacgc cccgcctgcc actcatcgca
      901 gtactgttgt aattcattaa gcattctgcc gacatggaag ccatcacaaa cggcatgatg
      961 aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt
     1021 gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact
     1081 cacccaggga ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc
     1141 caggttttca ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc
     1201 gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta
     1261 acaagggtga acactatccc atatcaccag ctcaccgtct ttcattgcca tacggaattc
     1321 cggatgagca ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt
     1381 atttttcttt acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt
     1441 acattgagcg actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc
     1501 aacggtggta tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct
     1561 cgacggatcc taactcaaaa tccacacatt atacgagccg gaagcataaa gtgtaaagcc
     1621 tggggtgcct aatgcggccg ccatagtgac tggatatgtt gtgttttaca gtattatgta
     1681 gtctgttttt tatgcaaaat ctaatttaat atattgatat ttatatcatt ttacgtttct
     1741 cgttcagctt ttttgtacaa acttgtcacc gatatccgtc tccgttagag ctcccggtga
     1801 cgcaggtgat gaaaagcctt agggtcaggt ggcacttttc ggggaaatgt gcgatgaacc
     1861 cctatttgtt tatttttcta aatacattca aatatgtatc cgctaatgag acaataaccc
     1921 tgataaatgc ttcaattata ttgaaaaagg aagagtatga gtatccaaca tttccgtgtc
     1981 gcccttattc ccttttttgc ggccttctgc cttcctgttt ttgctcaccc agaaacgctg
     2041 gtgaaagtaa aagatgctga agatcagttg ggtgcccgag tgggttacat tgaactggat
     2101 ctcaacagcg gcaaaatctt agagagtttt cgccccgaag aacgcttccc aatgatgagc
     2161 actttcaaag ttctgctatg tggcgcggtg ttatcccgta ttgatgctgg gcaagagcaa
     2221 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtattcacc agtcacagaa
     2281 aagcatctta cggatggcat gacagtaaga gaattatgta gtgctgccat aaccatgagt
     2341 gataacactg cggccaactt acttctgaca acgattggag gaccgaagga gctaaccgct
     2401 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     2461 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaactctc
     2521 cgcaaactat taactggcga actacttact cttgcttcac gccagcaact cattgactgg
     2581 atggaggcgg acaaagttgc aggaccactt ctgcgctcgg cacttccggc tggctggttt
     2641 attgctgata aatctggagc cggtgagcgt ggctctcgcg gtatcattgc agcactgggg
     2701 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     2761 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     2821 tcagacgaag ggaaataaat agtagcccgc ctgatgtgcg ggcttttttt tggtggtggt
     2881 taccagtgtc gcttagctac cgtaggtcct cttaggcggt ccgattagcg acatagtcat
     2941 caatgccttg acggcaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     3001 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3061 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctactaact
     3121 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     3181 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3241 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3301 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgctca
     3361 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     3421 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3481 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     3541 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     3601 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     3661 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     3721 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     3781 g
//
