LOCUS       sequence-334503-        8110 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid Double Transcriptional Unit - 
            p35SoTMV_E-Luc_AtACT2t+pNOS_Red-F_NOSt.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8110)
  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 8110)
  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: pGREAT12
FEATURES             Location/Qualifiers
     source          1..8110
                     /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        2842..3021
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      4726..4978
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(5013..5031)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5220..5244
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5335..5923)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5415..5434)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6094..6885)
                     /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      7529..7964
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    join(8091..8110,1..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 cttgctctca
     2101 agatcaaagg cttaaaaagc tggggtttta tgaatgggat caaagtttct ttttttcttt
     2161 tatatttgct tctccatttg tttgtttcat ttcccttttt gttttcgttt ctatgatgca
     2221 cttgtgtgtg acaaactctc tgggttttta cttacgtctg cgtttcaaaa aaaaaaaccg
     2281 ctttcgtttt gcgttttagt cccattgttt tgtagctctg agtgatcgaa ttgatgcctc
     2341 tttattcctt ttgttcccta taatttcttt caaaactcag aagaaaaacc ttgaaactct
     2401 ttgcaatgtt aatataagta ttgtataaga tttttattga tttggttatt agtcttactt
     2461 ttgctacctc catcttcact tggaactgat attctgaata gttaaagcgt tacatgtcct
     2521 ccattcacaa atgaacttaa actagcacaa agtcagatat tttaagatcg caccatttta
     2581 tataacccca atcgtcaatt ctactgtttc aagttttaca ccaaaacaat tacgaggtgt
     2641 atctattcgt tactcttttc gcttattaat ccttttacat gatgcaaaat ataacaaatt
     2701 aatttcaaat tgtcctgcac atttggttta tacatcttga ttccaaaaag tcgctgtcag
     2761 gagatacatg agaattaagg gagtcacggt atgacccccg ccgatgacgc gggacaagcc
     2821 gttttacgtt tggaactgac agaaccgcaa cgttgaagga gccactgagc cgcgggtttc
     2881 tggagtttaa tgagctaagc acatacgtca gaaaccatta ttgcgcgttc aaaagtcgcc
     2941 taaggtcact atcagctagc aaatatttct tgtcaaaaat gctccactga cgttccataa
     3001 attcccctcg gtatccaatt agagtctcat attcactctc aactcgatcg aggcaatgga
     3061 gaacatggag aacgacgaga acattgtggt gggacccaag ccgttctacc cgatcgagga
     3121 gggttccgct ggcacccagc tgcgcaagta catggagcgc tacgctaagc tgggtgctat
     3181 cgccttcacc aacgccgtga ccggtgtgga ttactcctac gccgagtacc tggagaagtc
     3241 ctgctgcctc ggaaaggccc tgcagaacta cggtctggtc gtggatggcc gcattgccct
     3301 ctgctcggag aactgcgagg agttcttcat tcccgtgatc gctggtctgt tcatcggagt
     3361 cggcgttgcc ccaaccaacg agatctacac cctgcgcgag ctggtccaca gcctgggtat
     3421 cagcaagccg accatcgttt tctcgagcaa gaagggcctc gacaaggtta tcaccgtgca
     3481 aaaaaccgtc accaccatca agaccatcgt tatcctggat tccaaggttg actaccgcgg
     3541 ttaccagtgc ctcgacacct tcatcaagcg taacacccca cccggcttcc aagcctcctc
     3601 cttcaaaacc gtggaggttg atcgcaagga gcaggtggcc ctgatcatga actcctccgg
     3661 ctccaccggt ctgccaaagg gcgtgcagct gacccacgag aacaccgtca cccgcttctc
     3721 gcatgcccgt gatcccatct acggcaacca ggtgtcccca ggcaccgccg tcctgaccgt
     3781 cgtgcccctc caccacggct tcggcatgtt caccactctc ggctacttga tctgcggctt
     3841 ccgtgtggtg atgctgacta agttcgacga ggagactttc ctcaagaccc tgcaagacta
     3901 caagtgcact tacgtgatcc tggtgccgac cctgttcgcc atcctgaaca agagcgagct
     3961 gttgaacaaa tacgacctgt cgaacctggt ggagatcgct tccggcggcg ccccactgtc
     4021 gaaggaagtt ggcgaggccg ttgctcgccg cttcaacctg cccggcgtcc gtcagggtta
     4081 cggcctgacc gagactacct cggctatcat catcaccccc gagggcgacg ataagcccgg
     4141 cgcttccggc aaggttgttc cactcttcaa agccaaggtg atcgacctgg acaccaagaa
     4201 gtccctgggt cccaaccgtc gcggagaagt gtgcgtcaag ggccccatgc tgatgaaggg
     4261 ctacgtcaac aacccagagg ccaccaagga gttgatcgac gaggagggct ggctgcacac
     4321 tggtgacatc ggttactacg acgaagagaa gcacttcttc atcgttgacc gcctgaagtc
     4381 cttgatcaag tacaagggct accaggttcc ccccgccgag ctcgagagcg tcctgctgca
     4441 gcacccctcc attttcgatg ccggagtggc cggtgtcccc gatcccgtgg ctggcgagct
     4501 gccaggtgcc gtcgtcgtgc tggagtccgg caagaacatg accgagaagg aagtgatgga
     4561 ttacgtggcc tcgcaagtgt ccaacgccaa gcgcctgcgc ggcggcgtgc gcttcgtgga
     4621 tgaggtgccc aagggcctca ccggaaagat cgacggtcgc gctatccgcg agatcctcaa
     4681 gaagcccgtg gccaagatgt gagcttggaa tggatcttcg atcccgatcg ttcaaacatt
     4741 tggcaataaa gtttcttaag attgaatcct gttgccggtc ttgcgacgat tatcatataa
     4801 tttctgttga attacgttaa gcatgtaata attaacatgt aatgcatgac gttatttatg
     4861 agatgggttt ttatgattag agtcccgcaa ttatacattt aatacgcgat agaaaacaaa
     4921 atatagcgcg caaactagga taaattatcg cgcgcggtgt catctatgtt actagatcgg
     4981 gaattgccaa gctaattctt gaagatgaaa gggcctcgtg atacgcctat ttttataggt
     5041 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     5101 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     5161 ataaccctga taaatgcttc aataatggga ccgactcgcg ctgtcatgag accggatcct
     5221 gacaggatat attggcgggt aaactaagtc gctgtatgtg tttgtttgag atctcatgtg
     5281 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5341 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5401 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5461 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5521 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5581 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5641 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5701 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5761 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5821 aagaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5881 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5941 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6001 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6061 ctaaagtata tatgtgtaac attggtctag tgattatttg ccgactacct tggtgatctc
     6121 gcctttcacg tagtggacaa attcttccaa ctgatctgcg cgcgaggcca agcgatcttc
     6181 ttcttgtcca agataagcct gtctagcttc aagtatgacg ggctgatact gggccggcag
     6241 gcgctccatt gcccagtcgg cagcgacatc cttcggcgcg attttgccgg ttactgcgct
     6301 gtaccaaatg cgggacaacg taagcactac atttcgctca tcaccagccc agtcgggcgg
     6361 cgagttccat agcgttaagg tttcatttag cgcctcaaat agatcctgtt caggaaccgg
     6421 atcaaagagt tcctccgccg ctggacctac caaggcaacg ctatgttctc ttgcttttgt
     6481 cagcaagata gccagatcaa tgtcgatcgt ggctggctcg aagatacctg caagaatgtc
     6541 attgcgctgc cattctccaa attgcagttc gcgcttagct ggataacgcc acggaatgat
     6601 gtcgtcgtgc acaacaatgg tgacttctac agcgcggaga atctcgctct ctccagggga
     6661 agccgaagtt tccaaaaggt cgttgatcaa agctcgccgc gttgtttcat caagccttac
     6721 ggtcaccgta accagcaaat caatatcact gtgtggcttc aggccgccat ccactgcgga
     6781 gccgtacaaa tgtacggcca gcaacgtcgg ttcgagatgg cgctcgatga cgccaactac
     6841 ctctgatagt tgagtcgata cttcggcgat caccgcttcc ctcataatgt ttaactttgt
     6901 tttagggcga ctgccctgct gcgtaacatc gttgctgctc cataacatca aacatcgacc
     6961 cacggcgtaa cgcgcttgct gcttggatgc ccgaggcata gactgtaccc caaaaaaaca
     7021 gtcataacaa gccatgaaaa ccgccactgc gccgttacca ccgctgcgtt cggtcaaggt
     7081 tctggaccag ttgcgtgagc gcatacgcta cttgcattac agcttacgaa ccgaacaggc
     7141 ttatgtccac tgggttcgtg ccttcatccg tttccacggt gtgcgtcacc cggcaacctt
     7201 gggtagcagc gaagtcgagg catttctgtc ctggctggaa caccccttgt attactgttt
     7261 atgtaagcag acagttttat tgttcatgat gatatatttt tatcttgtgc aatgtaacat
     7321 cagagatttt gagacacaac gtggctttgt tgaataaatc gaacttttgc tgagttgaag
     7381 gatcagatca cgcatcttcc cgacaacgca gaccgttccg tggcaaagca aaagttcaaa
     7441 atcaccaact ggtccaccta caacaaagct ctcatcaacc gtggctccct cactttctgg
     7501 ctggatgatg gggcgattca ggcgatcccc atccaacagc ccgccgtcga gcgggctttt
     7561 ttatccccgg aagcctgtgg atagagggta gttatccacg tgaaaccgct aatgccccgc
     7621 aaagccttga ttcacggggc tttccggccc gctccaaaaa ctatccacgt gaaatcgcta
     7681 atcagggtac gtgaaatcgc taatcggagt acgtgaaatc gctaataagg tcacgtgaaa
     7741 tcgctaatca aaaaggcacg tgagaacgct aatagccctt tcagatcaac agcttgcaaa
     7801 cacccctcgc tccggcaagt agttacagca agtagtatgt tcaattagct tttcaattat
     7861 gaatatatat atcaattatt ggtcgccctt ggcttgtgga caatgcgcta cgcgcaccgg
     7921 ctccgcccgt ggacaaccgc aagcggttgc ccaccgtcga gcgcctttgc ccacaacccg
     7981 gcggccggcc gcaacagatc gttttataaa tttttttttt tgaaaaagaa aaagcccgaa
     8041 aggcggcaac ctctcgggct tctggatttc cgatccccgg aattagatct tggcaggata
     8101 tattgtggtg
//
