LOCUS       sequence-334551-        7531 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid Double Transcriptional Unit - 
            pNOS_E-Luc_35St+pNOS_Red-F_NOSt.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7531)
  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 7531)
  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: pGREAT21
FEATURES             Location/Qualifiers
     source          1..7531
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        100..279
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     RBS             1786..1794
                     /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        2263..2442
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      4147..4399
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4434..4452)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4641..4665
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4756..5344)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4836..4855)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5515..6306)
                     /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      6950..7385
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    join(7512..7531,1..3)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
ORIGIN
        1 taacggatcc ggtctcagga gatacatgag aattaaggga gtcacggtat gacccccgcc
       61 gatgacgcgg gacaagccgt tttacgtttg gaactgacag aaccgcaacg ttgaaggagc
      121 cactgagccg cgggtttctg gagtttaatg agctaagcac atacgtcaga aaccattatt
      181 gcgcgttcaa aagtcgccta aggtcactat cagctagcaa atatttcttg tcaaaaatgc
      241 tccactgacg ttccataaat tcccctcggt atccaattag agtctcatat tcactctcaa
      301 ctcgatcgag gcaatggaga gagagaagaa cgtggtgtac ggccccgagc ccaagcaccc
      361 tctgggcaac ttcaccgccg gcgagatgct gtacaacgct ctgcacaagc actcccacat
      421 cccccaggcc atcctggacg tgatgggcaa cgagtccctt tcctaccagg agttcttcga
      481 cactactgtg aagctgggcc agagcctcca gaactgtggc tacaagatga acgatgtcgt
      541 gtcgatctgt gcagagaaca acaagagatt cttcatcccc atcatctccg cctggtacat
      601 cggcatggtg gtggcccctg tgaacgagga ctatatccca gacgagctgt gtaaagtgac
      661 cggcatctcc aagccgatcc tggtctttac cactaggaag atcctgccta aggttttgga
      721 ggttaaagac agaaccaact acataaagag gatcatcata ctggactctg aagagaacct
      781 gctgggctgc gagagcctgc acaacttcat gtccaggtac tccgacaaca acctccaaac
      841 attcaagcct ctgcactacg accctgtgga ccaggtagcc gccatcctgt gctcctccgg
      901 cacaaccggc ctgcctaaag gcgtgatgca gacccacagg aacatctgtg tgagactcac
      961 acacgcatct gaccccagag tgggtacaca actcatcccc ggcgtatccg tgctggccta
     1021 cctgccattc ttccacgcct tcggcttcag tatcaacctg ggctatttca tggtgggcct
     1081 gagagtggtg atgctccgaa ggtttaacca ggaggtgttc ctgaaggcca tccaggacta
     1141 cgaggtgagg agcgtgatca acgttccctc cacaatcctg ttcctgtcca agagccctct
     1201 ggtggacaag tacgacctat ccaccctggc ggagctgtgc tgtggagccg ctcctctggc
     1261 gaaggaggtg gccgagatcg ccgtgaagag gctgaacctg ccagggatac ggtgtggcta
     1321 cggtctaaca gagtctacct ccgccaacat ccatactctg cacaacgagt tcaagtccgg
     1381 ctccctgggc aaggtgacac cttacatggc cgccaagatc atcgacagga acaccggcga
     1441 ggccctgggt ccaaaccagg tgggcgagct gtgcatctgg ggacctatgg taacaaaagg
     1501 ctatgtgaac aacccacagg ctactaagga ggccatcgac gacgacggct ggctgcactc
     1561 tggcgacttc ggctactacg acgaggacga gtatttctac atcgtggacc ggtacaagga
     1621 gctgatcaaa tacaagggct atcaggtcgc ccctgtggag ctggaggaga tcctccttca
     1681 gcacccaggc atcagggacg tggccgtcgt gggtatccct gacatcgagg ccggcgagct
     1741 gccagccggc ttcgtggtga agcagcccgg cgcccaactc accgctaagg aggtgtacga
     1801 cttcctggcc cagagggtgt ctcactccaa gtacctgagg ggcggcgtaa ggttcgtgga
     1861 ctctatcccc aggaacgtga caggcaagat tagtcgaaaa gagctgaggg aggccctgat
     1921 ggagaaggct tctaagctgt aagcttcggc catgctagag tccgcaaaaa tcaccagtct
     1981 ctctctacaa atctatctct ctctattttt ctccagaata atgtgtgagt agttcccaga
     2041 taagggaatt agggttctta tagggtttcg ctcatgtgtt gagcatataa gaaaccctta
     2101 gtatgtattt gtatttgtaa aatacttcta tcaataaaat ttctaattcc taaaaccaaa
     2161 atccagtgac ctcgctgtca ggagatacat gagaattaag ggagtcacgg tatgaccccc
     2221 gccgatgacg cgggacaagc cgttttacgt ttggaactga cagaaccgca acgttgaagg
     2281 agccactgag ccgcgggttt ctggagttta atgagctaag cacatacgtc agaaaccatt
     2341 attgcgcgtt caaaagtcgc ctaaggtcac tatcagctag caaatatttc ttgtcaaaaa
     2401 tgctccactg acgttccata aattcccctc ggtatccaat tagagtctca tattcactct
     2461 caactcgatc gaggcaatgg agaacatgga gaacgacgag aacattgtgg tgggacccaa
     2521 gccgttctac ccgatcgagg agggttccgc tggcacccag ctgcgcaagt acatggagcg
     2581 ctacgctaag ctgggtgcta tcgccttcac caacgccgtg accggtgtgg attactccta
     2641 cgccgagtac ctggagaagt cctgctgcct cggaaaggcc ctgcagaact acggtctggt
     2701 cgtggatggc cgcattgccc tctgctcgga gaactgcgag gagttcttca ttcccgtgat
     2761 cgctggtctg ttcatcggag tcggcgttgc cccaaccaac gagatctaca ccctgcgcga
     2821 gctggtccac agcctgggta tcagcaagcc gaccatcgtt ttctcgagca agaagggcct
     2881 cgacaaggtt atcaccgtgc aaaaaaccgt caccaccatc aagaccatcg ttatcctgga
     2941 ttccaaggtt gactaccgcg gttaccagtg cctcgacacc ttcatcaagc gtaacacccc
     3001 acccggcttc caagcctcct ccttcaaaac cgtggaggtt gatcgcaagg agcaggtggc
     3061 cctgatcatg aactcctccg gctccaccgg tctgccaaag ggcgtgcagc tgacccacga
     3121 gaacaccgtc acccgcttct cgcatgcccg tgatcccatc tacggcaacc aggtgtcccc
     3181 aggcaccgcc gtcctgaccg tcgtgcccct ccaccacggc ttcggcatgt tcaccactct
     3241 cggctacttg atctgcggct tccgtgtggt gatgctgact aagttcgacg aggagacttt
     3301 cctcaagacc ctgcaagact acaagtgcac ttacgtgatc ctggtgccga ccctgttcgc
     3361 catcctgaac aagagcgagc tgttgaacaa atacgacctg tcgaacctgg tggagatcgc
     3421 ttccggcggc gccccactgt cgaaggaagt tggcgaggcc gttgctcgcc gcttcaacct
     3481 gcccggcgtc cgtcagggtt acggcctgac cgagactacc tcggctatca tcatcacccc
     3541 cgagggcgac gataagcccg gcgcttccgg caaggttgtt ccactcttca aagccaaggt
     3601 gatcgacctg gacaccaaga agtccctggg tcccaaccgt cgcggagaag tgtgcgtcaa
     3661 gggccccatg ctgatgaagg gctacgtcaa caacccagag gccaccaagg agttgatcga
     3721 cgaggagggc tggctgcaca ctggtgacat cggttactac gacgaagaga agcacttctt
     3781 catcgttgac cgcctgaagt ccttgatcaa gtacaagggc taccaggttc cccccgccga
     3841 gctcgagagc gtcctgctgc agcacccctc cattttcgat gccggagtgg ccggtgtccc
     3901 cgatcccgtg gctggcgagc tgccaggtgc cgtcgtcgtg ctggagtccg gcaagaacat
     3961 gaccgagaag gaagtgatgg attacgtggc ctcgcaagtg tccaacgcca agcgcctgcg
     4021 cggcggcgtg cgcttcgtgg atgaggtgcc caagggcctc accggaaaga tcgacggtcg
     4081 cgctatccgc gagatcctca agaagcccgt ggccaagatg tgagcttgga atggatcttc
     4141 gatcccgatc gttcaaacat ttggcaataa agtttcttaa gattgaatcc tgttgccggt
     4201 cttgcgacga ttatcatata atttctgttg aattacgtta agcatgtaat aattaacatg
     4261 taatgcatga cgttatttat gagatgggtt tttatgatta gagtcccgca attatacatt
     4321 taatacgcga tagaaaacaa aatatagcgc gcaaactagg ataaattatc gcgcgcggtg
     4381 tcatctatgt tactagatcg ggaattgcca agctaattct tgaagatgaa agggcctcgt
     4441 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     4501 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4561 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatggg accgactcgc
     4621 gctgtcatga gaccggatcc tgacaggata tattggcggg taaactaagt cgctgtatgt
     4681 gtttgtttga gatctcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4741 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4801 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4861 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4921 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4981 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5041 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5101 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5161 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5221 gctgaagcca gttaccttcg gaagaagagt tggtagctct tgatccggca aacaaaccac
     5281 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5341 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5401 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5461 aaaatgaagt tttaaatcaa tctaaagtat atatgtgtaa cattggtcta gtgattattt
     5521 gccgactacc ttggtgatct cgcctttcac gtagtggaca aattcttcca actgatctgc
     5581 gcgcgaggcc aagcgatctt cttcttgtcc aagataagcc tgtctagctt caagtatgac
     5641 gggctgatac tgggccggca ggcgctccat tgcccagtcg gcagcgacat ccttcggcgc
     5701 gattttgccg gttactgcgc tgtaccaaat gcgggacaac gtaagcacta catttcgctc
     5761 atcaccagcc cagtcgggcg gcgagttcca tagcgttaag gtttcattta gcgcctcaaa
     5821 tagatcctgt tcaggaaccg gatcaaagag ttcctccgcc gctggaccta ccaaggcaac
     5881 gctatgttct cttgcttttg tcagcaagat agccagatca atgtcgatcg tggctggctc
     5941 gaagatacct gcaagaatgt cattgcgctg ccattctcca aattgcagtt cgcgcttagc
     6001 tggataacgc cacggaatga tgtcgtcgtg cacaacaatg gtgacttcta cagcgcggag
     6061 aatctcgctc tctccagggg aagccgaagt ttccaaaagg tcgttgatca aagctcgccg
     6121 cgttgtttca tcaagcctta cggtcaccgt aaccagcaaa tcaatatcac tgtgtggctt
     6181 caggccgcca tccactgcgg agccgtacaa atgtacggcc agcaacgtcg gttcgagatg
     6241 gcgctcgatg acgccaacta cctctgatag ttgagtcgat acttcggcga tcaccgcttc
     6301 cctcataatg tttaactttg ttttagggcg actgccctgc tgcgtaacat cgttgctgct
     6361 ccataacatc aaacatcgac ccacggcgta acgcgcttgc tgcttggatg cccgaggcat
     6421 agactgtacc ccaaaaaaac agtcataaca agccatgaaa accgccactg cgccgttacc
     6481 accgctgcgt tcggtcaagg ttctggacca gttgcgtgag cgcatacgct acttgcatta
     6541 cagcttacga accgaacagg cttatgtcca ctgggttcgt gccttcatcc gtttccacgg
     6601 tgtgcgtcac ccggcaacct tgggtagcag cgaagtcgag gcatttctgt cctggctgga
     6661 acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga tgatatattt
     6721 ttatcttgtg caatgtaaca tcagagattt tgagacacaa cgtggctttg ttgaataaat
     6781 cgaacttttg ctgagttgaa ggatcagatc acgcatcttc ccgacaacgc agaccgttcc
     6841 gtggcaaagc aaaagttcaa aatcaccaac tggtccacct acaacaaagc tctcatcaac
     6901 cgtggctccc tcactttctg gctggatgat ggggcgattc aggcgatccc catccaacag
     6961 cccgccgtcg agcgggcttt tttatccccg gaagcctgtg gatagagggt agttatccac
     7021 gtgaaaccgc taatgccccg caaagccttg attcacgggg ctttccggcc cgctccaaaa
     7081 actatccacg tgaaatcgct aatcagggta cgtgaaatcg ctaatcggag tacgtgaaat
     7141 cgctaataag gtcacgtgaa atcgctaatc aaaaaggcac gtgagaacgc taatagccct
     7201 ttcagatcaa cagcttgcaa acacccctcg ctccggcaag tagttacagc aagtagtatg
     7261 ttcaattagc ttttcaatta tgaatatata tatcaattat tggtcgccct tggcttgtgg
     7321 acaatgcgct acgcgcaccg gctccgcccg tggacaaccg caagcggttg cccaccgtcg
     7381 agcgcctttg cccacaaccc ggcggccggc cgcaacagat cgttttataa attttttttt
     7441 ttgaaaaaga aaaagcccga aaggcggcaa cctctcgggc ttctggattt ccgatccccg
     7501 gaattagatc ttggcaggat atattgtggt g
//
