LOCUS       sequence-335148-        7791 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid Double Transcriptional Unit - 
            pNOS_E-Luc_NtEUt+pNOS_Red-F_NOSt.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7791)
  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 7791)
  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: pGREAT6
FEATURES             Location/Qualifiers
     source          1..7791
                     /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        2523..2702
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      4407..4659
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4694..4712)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4901..4925
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5016..5604)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5096..5115)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5775..6566)
                     /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      7210..7645
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    join(7772..7791,1..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 aagcttaaag cagaatgctg agctaaaaga aaggcttttt
     1981 ccattttcga gagacaatga gaaaagaaga agaagaagaa gaagaagaag aagaagaaga
     2041 agaaaagagt aaataataaa gccccacagg aggcgaagtt cttgtagctc catgttatct
     2101 aagttattga tattgtttgc cctatatttt atttctgtca ttgtgtatgt tttgttcagt
     2161 ttcgatctcc ttgcaaaatg cagagattat gagatgaata aactaagtta tattattata
     2221 cgtgttaata ttctcctcct ctctctagct agccttttgt tttctctttt tcttatttga
     2281 ttttctttaa atcaatccat tttaggagag ggccagggag tgatccagca aaacatgaag
     2341 attagaagaa acttccctct tttttttcct gaaaacaatt taacgtcgag atttatctct
     2401 ttttgtaatg gaatcatttc tacagttatg accgctgtca ggagatacat gagaattaag
     2461 ggagtcacgg tatgaccccc gccgatgacg cgggacaagc cgttttacgt ttggaactga
     2521 cagaaccgca acgttgaagg agccactgag ccgcgggttt ctggagttta atgagctaag
     2581 cacatacgtc agaaaccatt attgcgcgtt caaaagtcgc ctaaggtcac tatcagctag
     2641 caaatatttc ttgtcaaaaa tgctccactg acgttccata aattcccctc ggtatccaat
     2701 tagagtctca tattcactct caactcgatc gaggcaatgg agaacatgga gaacgacgag
     2761 aacattgtgg tgggacccaa gccgttctac ccgatcgagg agggttccgc tggcacccag
     2821 ctgcgcaagt acatggagcg ctacgctaag ctgggtgcta tcgccttcac caacgccgtg
     2881 accggtgtgg attactccta cgccgagtac ctggagaagt cctgctgcct cggaaaggcc
     2941 ctgcagaact acggtctggt cgtggatggc cgcattgccc tctgctcgga gaactgcgag
     3001 gagttcttca ttcccgtgat cgctggtctg ttcatcggag tcggcgttgc cccaaccaac
     3061 gagatctaca ccctgcgcga gctggtccac agcctgggta tcagcaagcc gaccatcgtt
     3121 ttctcgagca agaagggcct cgacaaggtt atcaccgtgc aaaaaaccgt caccaccatc
     3181 aagaccatcg ttatcctgga ttccaaggtt gactaccgcg gttaccagtg cctcgacacc
     3241 ttcatcaagc gtaacacccc acccggcttc caagcctcct ccttcaaaac cgtggaggtt
     3301 gatcgcaagg agcaggtggc cctgatcatg aactcctccg gctccaccgg tctgccaaag
     3361 ggcgtgcagc tgacccacga gaacaccgtc acccgcttct cgcatgcccg tgatcccatc
     3421 tacggcaacc aggtgtcccc aggcaccgcc gtcctgaccg tcgtgcccct ccaccacggc
     3481 ttcggcatgt tcaccactct cggctacttg atctgcggct tccgtgtggt gatgctgact
     3541 aagttcgacg aggagacttt cctcaagacc ctgcaagact acaagtgcac ttacgtgatc
     3601 ctggtgccga ccctgttcgc catcctgaac aagagcgagc tgttgaacaa atacgacctg
     3661 tcgaacctgg tggagatcgc ttccggcggc gccccactgt cgaaggaagt tggcgaggcc
     3721 gttgctcgcc gcttcaacct gcccggcgtc cgtcagggtt acggcctgac cgagactacc
     3781 tcggctatca tcatcacccc cgagggcgac gataagcccg gcgcttccgg caaggttgtt
     3841 ccactcttca aagccaaggt gatcgacctg gacaccaaga agtccctggg tcccaaccgt
     3901 cgcggagaag tgtgcgtcaa gggccccatg ctgatgaagg gctacgtcaa caacccagag
     3961 gccaccaagg agttgatcga cgaggagggc tggctgcaca ctggtgacat cggttactac
     4021 gacgaagaga agcacttctt catcgttgac cgcctgaagt ccttgatcaa gtacaagggc
     4081 taccaggttc cccccgccga gctcgagagc gtcctgctgc agcacccctc cattttcgat
     4141 gccggagtgg ccggtgtccc cgatcccgtg gctggcgagc tgccaggtgc cgtcgtcgtg
     4201 ctggagtccg gcaagaacat gaccgagaag gaagtgatgg attacgtggc ctcgcaagtg
     4261 tccaacgcca agcgcctgcg cggcggcgtg cgcttcgtgg atgaggtgcc caagggcctc
     4321 accggaaaga tcgacggtcg cgctatccgc gagatcctca agaagcccgt ggccaagatg
     4381 tgagcttgga atggatcttc gatcccgatc gttcaaacat ttggcaataa agtttcttaa
     4441 gattgaatcc tgttgccggt cttgcgacga ttatcatata atttctgttg aattacgtta
     4501 agcatgtaat aattaacatg taatgcatga cgttatttat gagatgggtt tttatgatta
     4561 gagtcccgca attatacatt taatacgcga tagaaaacaa aatatagcgc gcaaactagg
     4621 ataaattatc gcgcgcggtg tcatctatgt tactagatcg ggaattgcca agctaattct
     4681 tgaagatgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     4741 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4801 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     4861 caataatggg accgactcgc gctgtcatga gaccggatcc tgacaggata tattggcggg
     4921 taaactaagt cgctgtatgt gtttgtttga gatctcatgt gagcaaaagg ccagcaaaag
     4981 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5041 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5101 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5161 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5221 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5281 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5341 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5401 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5461 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaagaagagt tggtagctct
     5521 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5581 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5641 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5701 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgtgtaa
     5761 cattggtcta gtgattattt gccgactacc ttggtgatct cgcctttcac gtagtggaca
     5821 aattcttcca actgatctgc gcgcgaggcc aagcgatctt cttcttgtcc aagataagcc
     5881 tgtctagctt caagtatgac gggctgatac tgggccggca ggcgctccat tgcccagtcg
     5941 gcagcgacat ccttcggcgc gattttgccg gttactgcgc tgtaccaaat gcgggacaac
     6001 gtaagcacta catttcgctc atcaccagcc cagtcgggcg gcgagttcca tagcgttaag
     6061 gtttcattta gcgcctcaaa tagatcctgt tcaggaaccg gatcaaagag ttcctccgcc
     6121 gctggaccta ccaaggcaac gctatgttct cttgcttttg tcagcaagat agccagatca
     6181 atgtcgatcg tggctggctc gaagatacct gcaagaatgt cattgcgctg ccattctcca
     6241 aattgcagtt cgcgcttagc tggataacgc cacggaatga tgtcgtcgtg cacaacaatg
     6301 gtgacttcta cagcgcggag aatctcgctc tctccagggg aagccgaagt ttccaaaagg
     6361 tcgttgatca aagctcgccg cgttgtttca tcaagcctta cggtcaccgt aaccagcaaa
     6421 tcaatatcac tgtgtggctt caggccgcca tccactgcgg agccgtacaa atgtacggcc
     6481 agcaacgtcg gttcgagatg gcgctcgatg acgccaacta cctctgatag ttgagtcgat
     6541 acttcggcga tcaccgcttc cctcataatg tttaactttg ttttagggcg actgccctgc
     6601 tgcgtaacat cgttgctgct ccataacatc aaacatcgac ccacggcgta acgcgcttgc
     6661 tgcttggatg cccgaggcat agactgtacc ccaaaaaaac agtcataaca agccatgaaa
     6721 accgccactg cgccgttacc accgctgcgt tcggtcaagg ttctggacca gttgcgtgag
     6781 cgcatacgct acttgcatta cagcttacga accgaacagg cttatgtcca ctgggttcgt
     6841 gccttcatcc gtttccacgg tgtgcgtcac ccggcaacct tgggtagcag cgaagtcgag
     6901 gcatttctgt cctggctgga acaccccttg tattactgtt tatgtaagca gacagtttta
     6961 ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt tgagacacaa
     7021 cgtggctttg ttgaataaat cgaacttttg ctgagttgaa ggatcagatc acgcatcttc
     7081 ccgacaacgc agaccgttcc gtggcaaagc aaaagttcaa aatcaccaac tggtccacct
     7141 acaacaaagc tctcatcaac cgtggctccc tcactttctg gctggatgat ggggcgattc
     7201 aggcgatccc catccaacag cccgccgtcg agcgggcttt tttatccccg gaagcctgtg
     7261 gatagagggt agttatccac gtgaaaccgc taatgccccg caaagccttg attcacgggg
     7321 ctttccggcc cgctccaaaa actatccacg tgaaatcgct aatcagggta cgtgaaatcg
     7381 ctaatcggag tacgtgaaat cgctaataag gtcacgtgaa atcgctaatc aaaaaggcac
     7441 gtgagaacgc taatagccct ttcagatcaa cagcttgcaa acacccctcg ctccggcaag
     7501 tagttacagc aagtagtatg ttcaattagc ttttcaatta tgaatatata tatcaattat
     7561 tggtcgccct tggcttgtgg acaatgcgct acgcgcaccg gctccgcccg tggacaaccg
     7621 caagcggttg cccaccgtcg agcgcctttg cccacaaccc ggcggccggc cgcaacagat
     7681 cgttttataa attttttttt ttgaaaaaga aaaagcccga aaggcggcaa cctctcgggc
     7741 ttctggattt ccgatccccg gaattagatc ttggcaggat atattgtggt g
//
