LOCUS       sequence-334498-        7938 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid Double Transcriptional Unit - 
            p35SoTMV_E-Luc_NtEUt+pNOS_Red-F_NOSt.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7938)
  AUTHORS   Gonzalez-Grandio E, Demirer GS, Ma W, Brady S, Landry MP
  TITLE     A Ratiometric Dual Color Luciferase Reporter for Fast 
            Characterization of Transcriptional Regulatory Elements in Plants.
  JOURNAL   ACS Synth Biol. 2021 Sep 14. doi: 10.1021/acssynbio.1c00248.
  PUBMED    34520169
REFERENCE   2  (bases 1 to 7938)
  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: "ACS Synth 
            Biol. 2021 Sep 14. doi: 10.1021/acssynbio.1c00248."
COMMENT     Sequence Label: pGREAT7
FEATURES             Location/Qualifiers
     source          1..7938
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        22..367
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     310..330
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    387..442
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     RBS             1933..1941
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     promoter        2670..2849
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      4554..4806
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4841..4859)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5048..5072
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5163..5751)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5243..5262)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5922..6713)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      7357..7792
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    join(7919..7938,1..3)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
ORIGIN
        1 taacggatcc ggtctcagga gtgagacttt tcaacaaagg gtaatatcgg gaaacctcct
       61 cggattccat tgcccagcta tctgtcactt catcgaaagg acagtagaaa aggaaggtgg
      121 ctcctacaaa tgccatcatt gcgataaagg aaaggctatc gttcaagatg cctctaccga
      181 cagtggtccc aaagatggac ccccacccac gaggaacatc gtggaaaaag aagatgttcc
      241 aaccacgtcc tcaaagcaag tggattgatg tgatatctcc actgacgtaa gggatgacgc
      301 acaatcccac tatccttcgc aagacccttc ctctatataa ggaagttcat ttcatttgga
      361 gaggacaggc ttcttgagat ccttcaacaa ttaccaacaa caacaaacaa caaacaacat
      421 tacaattact atttacaatt acagtcgact ctagaggata atggagagag agaagaacgt
      481 ggtgtacggc cccgagccca agcaccctct gggcaacttc accgccggcg agatgctgta
      541 caacgctctg cacaagcact cccacatccc ccaggccatc ctggacgtga tgggcaacga
      601 gtccctttcc taccaggagt tcttcgacac tactgtgaag ctgggccaga gcctccagaa
      661 ctgtggctac aagatgaacg atgtcgtgtc gatctgtgca gagaacaaca agagattctt
      721 catccccatc atctccgcct ggtacatcgg catggtggtg gcccctgtga acgaggacta
      781 tatcccagac gagctgtgta aagtgaccgg catctccaag ccgatcctgg tctttaccac
      841 taggaagatc ctgcctaagg ttttggaggt taaagacaga accaactaca taaagaggat
      901 catcatactg gactctgaag agaacctgct gggctgcgag agcctgcaca acttcatgtc
      961 caggtactcc gacaacaacc tccaaacatt caagcctctg cactacgacc ctgtggacca
     1021 ggtagccgcc atcctgtgct cctccggcac aaccggcctg cctaaaggcg tgatgcagac
     1081 ccacaggaac atctgtgtga gactcacaca cgcatctgac cccagagtgg gtacacaact
     1141 catccccggc gtatccgtgc tggcctacct gccattcttc cacgccttcg gcttcagtat
     1201 caacctgggc tatttcatgg tgggcctgag agtggtgatg ctccgaaggt ttaaccagga
     1261 ggtgttcctg aaggccatcc aggactacga ggtgaggagc gtgatcaacg ttccctccac
     1321 aatcctgttc ctgtccaaga gccctctggt ggacaagtac gacctatcca ccctggcgga
     1381 gctgtgctgt ggagccgctc ctctggcgaa ggaggtggcc gagatcgccg tgaagaggct
     1441 gaacctgcca gggatacggt gtggctacgg tctaacagag tctacctccg ccaacatcca
     1501 tactctgcac aacgagttca agtccggctc cctgggcaag gtgacacctt acatggccgc
     1561 caagatcatc gacaggaaca ccggcgaggc cctgggtcca aaccaggtgg gcgagctgtg
     1621 catctgggga cctatggtaa caaaaggcta tgtgaacaac ccacaggcta ctaaggaggc
     1681 catcgacgac gacggctggc tgcactctgg cgacttcggc tactacgacg aggacgagta
     1741 tttctacatc gtggaccggt acaaggagct gatcaaatac aagggctatc aggtcgcccc
     1801 tgtggagctg gaggagatcc tccttcagca cccaggcatc agggacgtgg ccgtcgtggg
     1861 tatccctgac atcgaggccg gcgagctgcc agccggcttc gtggtgaagc agcccggcgc
     1921 ccaactcacc gctaaggagg tgtacgactt cctggcccag agggtgtctc actccaagta
     1981 cctgaggggc ggcgtaaggt tcgtggactc tatccccagg aacgtgacag gcaagattag
     2041 tcgaaaagag ctgagggagg ccctgatgga gaaggcttct aagctgtaag cttaaagcag
     2101 aatgctgagc taaaagaaag gctttttcca ttttcgagag acaatgagaa aagaagaaga
     2161 agaagaagaa gaagaagaag aagaagaaga aaagagtaaa taataaagcc ccacaggagg
     2221 cgaagttctt gtagctccat gttatctaag ttattgatat tgtttgccct atattttatt
     2281 tctgtcattg tgtatgtttt gttcagtttc gatctccttg caaaatgcag agattatgag
     2341 atgaataaac taagttatat tattatacgt gttaatattc tcctcctctc tctagctagc
     2401 cttttgtttt ctctttttct tatttgattt tctttaaatc aatccatttt aggagagggc
     2461 cagggagtga tccagcaaaa catgaagatt agaagaaact tccctctttt ttttcctgaa
     2521 aacaatttaa cgtcgagatt tatctctttt tgtaatggaa tcatttctac agttatgacc
     2581 gctgtcagga gatacatgag aattaaggga gtcacggtat gacccccgcc gatgacgcgg
     2641 gacaagccgt tttacgtttg gaactgacag aaccgcaacg ttgaaggagc cactgagccg
     2701 cgggtttctg gagtttaatg agctaagcac atacgtcaga aaccattatt gcgcgttcaa
     2761 aagtcgccta aggtcactat cagctagcaa atatttcttg tcaaaaatgc tccactgacg
     2821 ttccataaat tcccctcggt atccaattag agtctcatat tcactctcaa ctcgatcgag
     2881 gcaatggaga acatggagaa cgacgagaac attgtggtgg gacccaagcc gttctacccg
     2941 atcgaggagg gttccgctgg cacccagctg cgcaagtaca tggagcgcta cgctaagctg
     3001 ggtgctatcg ccttcaccaa cgccgtgacc ggtgtggatt actcctacgc cgagtacctg
     3061 gagaagtcct gctgcctcgg aaaggccctg cagaactacg gtctggtcgt ggatggccgc
     3121 attgccctct gctcggagaa ctgcgaggag ttcttcattc ccgtgatcgc tggtctgttc
     3181 atcggagtcg gcgttgcccc aaccaacgag atctacaccc tgcgcgagct ggtccacagc
     3241 ctgggtatca gcaagccgac catcgttttc tcgagcaaga agggcctcga caaggttatc
     3301 accgtgcaaa aaaccgtcac caccatcaag accatcgtta tcctggattc caaggttgac
     3361 taccgcggtt accagtgcct cgacaccttc atcaagcgta acaccccacc cggcttccaa
     3421 gcctcctcct tcaaaaccgt ggaggttgat cgcaaggagc aggtggccct gatcatgaac
     3481 tcctccggct ccaccggtct gccaaagggc gtgcagctga cccacgagaa caccgtcacc
     3541 cgcttctcgc atgcccgtga tcccatctac ggcaaccagg tgtccccagg caccgccgtc
     3601 ctgaccgtcg tgcccctcca ccacggcttc ggcatgttca ccactctcgg ctacttgatc
     3661 tgcggcttcc gtgtggtgat gctgactaag ttcgacgagg agactttcct caagaccctg
     3721 caagactaca agtgcactta cgtgatcctg gtgccgaccc tgttcgccat cctgaacaag
     3781 agcgagctgt tgaacaaata cgacctgtcg aacctggtgg agatcgcttc cggcggcgcc
     3841 ccactgtcga aggaagttgg cgaggccgtt gctcgccgct tcaacctgcc cggcgtccgt
     3901 cagggttacg gcctgaccga gactacctcg gctatcatca tcacccccga gggcgacgat
     3961 aagcccggcg cttccggcaa ggttgttcca ctcttcaaag ccaaggtgat cgacctggac
     4021 accaagaagt ccctgggtcc caaccgtcgc ggagaagtgt gcgtcaaggg ccccatgctg
     4081 atgaagggct acgtcaacaa cccagaggcc accaaggagt tgatcgacga ggagggctgg
     4141 ctgcacactg gtgacatcgg ttactacgac gaagagaagc acttcttcat cgttgaccgc
     4201 ctgaagtcct tgatcaagta caagggctac caggttcccc ccgccgagct cgagagcgtc
     4261 ctgctgcagc acccctccat tttcgatgcc ggagtggccg gtgtccccga tcccgtggct
     4321 ggcgagctgc caggtgccgt cgtcgtgctg gagtccggca agaacatgac cgagaaggaa
     4381 gtgatggatt acgtggcctc gcaagtgtcc aacgccaagc gcctgcgcgg cggcgtgcgc
     4441 ttcgtggatg aggtgcccaa gggcctcacc ggaaagatcg acggtcgcgc tatccgcgag
     4501 atcctcaaga agcccgtggc caagatgtga gcttggaatg gatcttcgat cccgatcgtt
     4561 caaacatttg gcaataaagt ttcttaagat tgaatcctgt tgccggtctt gcgacgatta
     4621 tcatataatt tctgttgaat tacgttaagc atgtaataat taacatgtaa tgcatgacgt
     4681 tatttatgag atgggttttt atgattagag tcccgcaatt atacatttaa tacgcgatag
     4741 aaaacaaaat atagcgcgca aactaggata aattatcgcg cgcggtgtca tctatgttac
     4801 tagatcggga attgccaagc taattcttga agatgaaagg gcctcgtgat acgcctattt
     4861 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     4921 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4981 atgagacaat aaccctgata aatgcttcaa taatgggacc gactcgcgct gtcatgagac
     5041 cggatcctga caggatatat tggcgggtaa actaagtcgc tgtatgtgtt tgtttgagat
     5101 ctcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5161 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5221 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5281 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5341 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5401 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5461 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5521 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5581 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     5641 accttcggaa gaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5701 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5761 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5821 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5881 aaatcaatct aaagtatata tgtgtaacat tggtctagtg attatttgcc gactaccttg
     5941 gtgatctcgc ctttcacgta gtggacaaat tcttccaact gatctgcgcg cgaggccaag
     6001 cgatcttctt cttgtccaag ataagcctgt ctagcttcaa gtatgacggg ctgatactgg
     6061 gccggcaggc gctccattgc ccagtcggca gcgacatcct tcggcgcgat tttgccggtt
     6121 actgcgctgt accaaatgcg ggacaacgta agcactacat ttcgctcatc accagcccag
     6181 tcgggcggcg agttccatag cgttaaggtt tcatttagcg cctcaaatag atcctgttca
     6241 ggaaccggat caaagagttc ctccgccgct ggacctacca aggcaacgct atgttctctt
     6301 gcttttgtca gcaagatagc cagatcaatg tcgatcgtgg ctggctcgaa gatacctgca
     6361 agaatgtcat tgcgctgcca ttctccaaat tgcagttcgc gcttagctgg ataacgccac
     6421 ggaatgatgt cgtcgtgcac aacaatggtg acttctacag cgcggagaat ctcgctctct
     6481 ccaggggaag ccgaagtttc caaaaggtcg ttgatcaaag ctcgccgcgt tgtttcatca
     6541 agccttacgg tcaccgtaac cagcaaatca atatcactgt gtggcttcag gccgccatcc
     6601 actgcggagc cgtacaaatg tacggccagc aacgtcggtt cgagatggcg ctcgatgacg
     6661 ccaactacct ctgatagttg agtcgatact tcggcgatca ccgcttccct cataatgttt
     6721 aactttgttt tagggcgact gccctgctgc gtaacatcgt tgctgctcca taacatcaaa
     6781 catcgaccca cggcgtaacg cgcttgctgc ttggatgccc gaggcataga ctgtacccca
     6841 aaaaaacagt cataacaagc catgaaaacc gccactgcgc cgttaccacc gctgcgttcg
     6901 gtcaaggttc tggaccagtt gcgtgagcgc atacgctact tgcattacag cttacgaacc
     6961 gaacaggctt atgtccactg ggttcgtgcc ttcatccgtt tccacggtgt gcgtcacccg
     7021 gcaaccttgg gtagcagcga agtcgaggca tttctgtcct ggctggaaca ccccttgtat
     7081 tactgtttat gtaagcagac agttttattg ttcatgatga tatattttta tcttgtgcaa
     7141 tgtaacatca gagattttga gacacaacgt ggctttgttg aataaatcga acttttgctg
     7201 agttgaagga tcagatcacg catcttcccg acaacgcaga ccgttccgtg gcaaagcaaa
     7261 agttcaaaat caccaactgg tccacctaca acaaagctct catcaaccgt ggctccctca
     7321 ctttctggct ggatgatggg gcgattcagg cgatccccat ccaacagccc gccgtcgagc
     7381 gggctttttt atccccggaa gcctgtggat agagggtagt tatccacgtg aaaccgctaa
     7441 tgccccgcaa agccttgatt cacggggctt tccggcccgc tccaaaaact atccacgtga
     7501 aatcgctaat cagggtacgt gaaatcgcta atcggagtac gtgaaatcgc taataaggtc
     7561 acgtgaaatc gctaatcaaa aaggcacgtg agaacgctaa tagccctttc agatcaacag
     7621 cttgcaaaca cccctcgctc cggcaagtag ttacagcaag tagtatgttc aattagcttt
     7681 tcaattatga atatatatat caattattgg tcgcccttgg cttgtggaca atgcgctacg
     7741 cgcaccggct ccgcccgtgg acaaccgcaa gcggttgccc accgtcgagc gcctttgccc
     7801 acaacccggc ggccggccgc aacagatcgt tttataaatt tttttttttg aaaaagaaaa
     7861 agcccgaaag gcggcaacct ctcgggcttc tggatttccg atccccggaa ttagatcttg
     7921 gcaggatata ttgtggtg
//
