LOCUS       sequence-334486-        7555 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid Double Transcriptional Unit - 
            pNOS_E-Luc_AtHSP18.2t+pNOS_Red-F_NOSt.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7555)
  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 7555)
  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: pGREAT16
FEATURES             Location/Qualifiers
     source          1..7555
                     /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)"
     terminator      1948..2196
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     promoter        2287..2466
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      4171..4423
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4458..4476)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4665..4689
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4780..5368)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4860..4879)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5539..6330)
                     /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      6974..7409
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    join(7536..7555,1..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 aagcttatat gaagatgaag atgaaatatt tggtgtgtca
     1981 aataaaaagc ttgtgtgctt aagtttgtgt ttttttcttg gcttgttgtg ttatgaattt
     2041 gtggcttttt ctaatattaa atgaatgtaa gatctcatta taatgaataa acaaatgttt
     2101 ctataatcca ttgtgaatgt tttgttggat ctcttctgca gcatataact actgtatgtg
     2161 ctatggtatg gactatggaa tatgattaaa gataagcgct gtcaggagat acatgagaat
     2221 taagggagtc acggtatgac ccccgccgat gacgcgggac aagccgtttt acgtttggaa
     2281 ctgacagaac cgcaacgttg aaggagccac tgagccgcgg gtttctggag tttaatgagc
     2341 taagcacata cgtcagaaac cattattgcg cgttcaaaag tcgcctaagg tcactatcag
     2401 ctagcaaata tttcttgtca aaaatgctcc actgacgttc cataaattcc cctcggtatc
     2461 caattagagt ctcatattca ctctcaactc gatcgaggca atggagaaca tggagaacga
     2521 cgagaacatt gtggtgggac ccaagccgtt ctacccgatc gaggagggtt ccgctggcac
     2581 ccagctgcgc aagtacatgg agcgctacgc taagctgggt gctatcgcct tcaccaacgc
     2641 cgtgaccggt gtggattact cctacgccga gtacctggag aagtcctgct gcctcggaaa
     2701 ggccctgcag aactacggtc tggtcgtgga tggccgcatt gccctctgct cggagaactg
     2761 cgaggagttc ttcattcccg tgatcgctgg tctgttcatc ggagtcggcg ttgccccaac
     2821 caacgagatc tacaccctgc gcgagctggt ccacagcctg ggtatcagca agccgaccat
     2881 cgttttctcg agcaagaagg gcctcgacaa ggttatcacc gtgcaaaaaa ccgtcaccac
     2941 catcaagacc atcgttatcc tggattccaa ggttgactac cgcggttacc agtgcctcga
     3001 caccttcatc aagcgtaaca ccccacccgg cttccaagcc tcctccttca aaaccgtgga
     3061 ggttgatcgc aaggagcagg tggccctgat catgaactcc tccggctcca ccggtctgcc
     3121 aaagggcgtg cagctgaccc acgagaacac cgtcacccgc ttctcgcatg cccgtgatcc
     3181 catctacggc aaccaggtgt ccccaggcac cgccgtcctg accgtcgtgc ccctccacca
     3241 cggcttcggc atgttcacca ctctcggcta cttgatctgc ggcttccgtg tggtgatgct
     3301 gactaagttc gacgaggaga ctttcctcaa gaccctgcaa gactacaagt gcacttacgt
     3361 gatcctggtg ccgaccctgt tcgccatcct gaacaagagc gagctgttga acaaatacga
     3421 cctgtcgaac ctggtggaga tcgcttccgg cggcgcccca ctgtcgaagg aagttggcga
     3481 ggccgttgct cgccgcttca acctgcccgg cgtccgtcag ggttacggcc tgaccgagac
     3541 tacctcggct atcatcatca cccccgaggg cgacgataag cccggcgctt ccggcaaggt
     3601 tgttccactc ttcaaagcca aggtgatcga cctggacacc aagaagtccc tgggtcccaa
     3661 ccgtcgcgga gaagtgtgcg tcaagggccc catgctgatg aagggctacg tcaacaaccc
     3721 agaggccacc aaggagttga tcgacgagga gggctggctg cacactggtg acatcggtta
     3781 ctacgacgaa gagaagcact tcttcatcgt tgaccgcctg aagtccttga tcaagtacaa
     3841 gggctaccag gttccccccg ccgagctcga gagcgtcctg ctgcagcacc cctccatttt
     3901 cgatgccgga gtggccggtg tccccgatcc cgtggctggc gagctgccag gtgccgtcgt
     3961 cgtgctggag tccggcaaga acatgaccga gaaggaagtg atggattacg tggcctcgca
     4021 agtgtccaac gccaagcgcc tgcgcggcgg cgtgcgcttc gtggatgagg tgcccaaggg
     4081 cctcaccgga aagatcgacg gtcgcgctat ccgcgagatc ctcaagaagc ccgtggccaa
     4141 gatgtgagct tggaatggat cttcgatccc gatcgttcaa acatttggca ataaagtttc
     4201 ttaagattga atcctgttgc cggtcttgcg acgattatca tataatttct gttgaattac
     4261 gttaagcatg taataattaa catgtaatgc atgacgttat ttatgagatg ggtttttatg
     4321 attagagtcc cgcaattata catttaatac gcgatagaaa acaaaatata gcgcgcaaac
     4381 taggataaat tatcgcgcgc ggtgtcatct atgttactag atcgggaatt gccaagctaa
     4441 ttcttgaaga tgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     4501 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4561 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4621 gcttcaataa tgggaccgac tcgcgctgtc atgagaccgg atcctgacag gatatattgg
     4681 cgggtaaact aagtcgctgt atgtgtttgt ttgagatctc atgtgagcaa aaggccagca
     4741 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4801 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4861 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4921 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4981 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5041 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5101 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5161 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5221 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaagaa gagttggtag
     5281 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5341 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5401 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5461 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatgt
     5521 gtaacattgg tctagtgatt atttgccgac taccttggtg atctcgcctt tcacgtagtg
     5581 gacaaattct tccaactgat ctgcgcgcga ggccaagcga tcttcttctt gtccaagata
     5641 agcctgtcta gcttcaagta tgacgggctg atactgggcc ggcaggcgct ccattgccca
     5701 gtcggcagcg acatccttcg gcgcgatttt gccggttact gcgctgtacc aaatgcggga
     5761 caacgtaagc actacatttc gctcatcacc agcccagtcg ggcggcgagt tccatagcgt
     5821 taaggtttca tttagcgcct caaatagatc ctgttcagga accggatcaa agagttcctc
     5881 cgccgctgga cctaccaagg caacgctatg ttctcttgct tttgtcagca agatagccag
     5941 atcaatgtcg atcgtggctg gctcgaagat acctgcaaga atgtcattgc gctgccattc
     6001 tccaaattgc agttcgcgct tagctggata acgccacgga atgatgtcgt cgtgcacaac
     6061 aatggtgact tctacagcgc ggagaatctc gctctctcca ggggaagccg aagtttccaa
     6121 aaggtcgttg atcaaagctc gccgcgttgt ttcatcaagc cttacggtca ccgtaaccag
     6181 caaatcaata tcactgtgtg gcttcaggcc gccatccact gcggagccgt acaaatgtac
     6241 ggccagcaac gtcggttcga gatggcgctc gatgacgcca actacctctg atagttgagt
     6301 cgatacttcg gcgatcaccg cttccctcat aatgtttaac tttgttttag ggcgactgcc
     6361 ctgctgcgta acatcgttgc tgctccataa catcaaacat cgacccacgg cgtaacgcgc
     6421 ttgctgcttg gatgcccgag gcatagactg taccccaaaa aaacagtcat aacaagccat
     6481 gaaaaccgcc actgcgccgt taccaccgct gcgttcggtc aaggttctgg accagttgcg
     6541 tgagcgcata cgctacttgc attacagctt acgaaccgaa caggcttatg tccactgggt
     6601 tcgtgccttc atccgtttcc acggtgtgcg tcacccggca accttgggta gcagcgaagt
     6661 cgaggcattt ctgtcctggc tggaacaccc cttgtattac tgtttatgta agcagacagt
     6721 tttattgttc atgatgatat atttttatct tgtgcaatgt aacatcagag attttgagac
     6781 acaacgtggc tttgttgaat aaatcgaact tttgctgagt tgaaggatca gatcacgcat
     6841 cttcccgaca acgcagaccg ttccgtggca aagcaaaagt tcaaaatcac caactggtcc
     6901 acctacaaca aagctctcat caaccgtggc tccctcactt tctggctgga tgatggggcg
     6961 attcaggcga tccccatcca acagcccgcc gtcgagcggg cttttttatc cccggaagcc
     7021 tgtggataga gggtagttat ccacgtgaaa ccgctaatgc cccgcaaagc cttgattcac
     7081 ggggctttcc ggcccgctcc aaaaactatc cacgtgaaat cgctaatcag ggtacgtgaa
     7141 atcgctaatc ggagtacgtg aaatcgctaa taaggtcacg tgaaatcgct aatcaaaaag
     7201 gcacgtgaga acgctaatag ccctttcaga tcaacagctt gcaaacaccc ctcgctccgg
     7261 caagtagtta cagcaagtag tatgttcaat tagcttttca attatgaata tatatatcaa
     7321 ttattggtcg cccttggctt gtggacaatg cgctacgcgc accggctccg cccgtggaca
     7381 accgcaagcg gttgcccacc gtcgagcgcc tttgcccaca acccggcggc cggccgcaac
     7441 agatcgtttt ataaattttt ttttttgaaa aagaaaaagc ccgaaaggcg gcaacctctc
     7501 gggcttctgg atttccgatc cccggaatta gatcttggca ggatatattg tggtg
//
