LOCUS       sequence-334550-        7743 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid Double Transcriptional Unit - 
            pG10-90_E-Luc_NtEUt+pNOS_Red-F_NOSt.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7743)
  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 7743)
  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: pGREAT9
FEATURES             Location/Qualifiers
     source          1..7743
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     196..216
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     promoter        205..251
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     RBS             1738..1746
                     /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        2475..2654
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      4359..4611
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4646..4664)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4853..4877
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4968..5556)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5048..5067)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5727..6518)
                     /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      7162..7597
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    join(7724..7743,1..3)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
ORIGIN
        1 taacggatcc ggtctcagga gatagtttaa actgaaggcg ggaaacgaca atctgatcca
       61 agctcaagct aagcttgcat gcctgcagga tatcgtggat ccaagcttgc cacgtgccgc
      121 cacgtgccgc cacgtgccgc cacgtgcctc tagaggatcc atctccactg acgtaaggga
      181 tgacgcacaa tcccactatc cttcgcaaga cccttcctct atataaggaa gttcatttca
      241 tttggagagg acacgctggg atccaatgga gagagagaag aacgtggtgt acggccccga
      301 gcccaagcac cctctgggca acttcaccgc cggcgagatg ctgtacaacg ctctgcacaa
      361 gcactcccac atcccccagg ccatcctgga cgtgatgggc aacgagtccc tttcctacca
      421 ggagttcttc gacactactg tgaagctggg ccagagcctc cagaactgtg gctacaagat
      481 gaacgatgtc gtgtcgatct gtgcagagaa caacaagaga ttcttcatcc ccatcatctc
      541 cgcctggtac atcggcatgg tggtggcccc tgtgaacgag gactatatcc cagacgagct
      601 gtgtaaagtg accggcatct ccaagccgat cctggtcttt accactagga agatcctgcc
      661 taaggttttg gaggttaaag acagaaccaa ctacataaag aggatcatca tactggactc
      721 tgaagagaac ctgctgggct gcgagagcct gcacaacttc atgtccaggt actccgacaa
      781 caacctccaa acattcaagc ctctgcacta cgaccctgtg gaccaggtag ccgccatcct
      841 gtgctcctcc ggcacaaccg gcctgcctaa aggcgtgatg cagacccaca ggaacatctg
      901 tgtgagactc acacacgcat ctgaccccag agtgggtaca caactcatcc ccggcgtatc
      961 cgtgctggcc tacctgccat tcttccacgc cttcggcttc agtatcaacc tgggctattt
     1021 catggtgggc ctgagagtgg tgatgctccg aaggtttaac caggaggtgt tcctgaaggc
     1081 catccaggac tacgaggtga ggagcgtgat caacgttccc tccacaatcc tgttcctgtc
     1141 caagagccct ctggtggaca agtacgacct atccaccctg gcggagctgt gctgtggagc
     1201 cgctcctctg gcgaaggagg tggccgagat cgccgtgaag aggctgaacc tgccagggat
     1261 acggtgtggc tacggtctaa cagagtctac ctccgccaac atccatactc tgcacaacga
     1321 gttcaagtcc ggctccctgg gcaaggtgac accttacatg gccgccaaga tcatcgacag
     1381 gaacaccggc gaggccctgg gtccaaacca ggtgggcgag ctgtgcatct ggggacctat
     1441 ggtaacaaaa ggctatgtga acaacccaca ggctactaag gaggccatcg acgacgacgg
     1501 ctggctgcac tctggcgact tcggctacta cgacgaggac gagtatttct acatcgtgga
     1561 ccggtacaag gagctgatca aatacaaggg ctatcaggtc gcccctgtgg agctggagga
     1621 gatcctcctt cagcacccag gcatcaggga cgtggccgtc gtgggtatcc ctgacatcga
     1681 ggccggcgag ctgccagccg gcttcgtggt gaagcagccc ggcgcccaac tcaccgctaa
     1741 ggaggtgtac gacttcctgg cccagagggt gtctcactcc aagtacctga ggggcggcgt
     1801 aaggttcgtg gactctatcc ccaggaacgt gacaggcaag attagtcgaa aagagctgag
     1861 ggaggccctg atggagaagg cttctaagct gtaagcttaa agcagaatgc tgagctaaaa
     1921 gaaaggcttt ttccattttc gagagacaat gagaaaagaa gaagaagaag aagaagaaga
     1981 agaagaagaa gaagaaaaga gtaaataata aagccccaca ggaggcgaag ttcttgtagc
     2041 tccatgttat ctaagttatt gatattgttt gccctatatt ttatttctgt cattgtgtat
     2101 gttttgttca gtttcgatct ccttgcaaaa tgcagagatt atgagatgaa taaactaagt
     2161 tatattatta tacgtgttaa tattctcctc ctctctctag ctagcctttt gttttctctt
     2221 tttcttattt gattttcttt aaatcaatcc attttaggag agggccaggg agtgatccag
     2281 caaaacatga agattagaag aaacttccct cttttttttc ctgaaaacaa tttaacgtcg
     2341 agatttatct ctttttgtaa tggaatcatt tctacagtta tgaccgctgt caggagatac
     2401 atgagaatta agggagtcac ggtatgaccc ccgccgatga cgcgggacaa gccgttttac
     2461 gtttggaact gacagaaccg caacgttgaa ggagccactg agccgcgggt ttctggagtt
     2521 taatgagcta agcacatacg tcagaaacca ttattgcgcg ttcaaaagtc gcctaaggtc
     2581 actatcagct agcaaatatt tcttgtcaaa aatgctccac tgacgttcca taaattcccc
     2641 tcggtatcca attagagtct catattcact ctcaactcga tcgaggcaat ggagaacatg
     2701 gagaacgacg agaacattgt ggtgggaccc aagccgttct acccgatcga ggagggttcc
     2761 gctggcaccc agctgcgcaa gtacatggag cgctacgcta agctgggtgc tatcgccttc
     2821 accaacgccg tgaccggtgt ggattactcc tacgccgagt acctggagaa gtcctgctgc
     2881 ctcggaaagg ccctgcagaa ctacggtctg gtcgtggatg gccgcattgc cctctgctcg
     2941 gagaactgcg aggagttctt cattcccgtg atcgctggtc tgttcatcgg agtcggcgtt
     3001 gccccaacca acgagatcta caccctgcgc gagctggtcc acagcctggg tatcagcaag
     3061 ccgaccatcg ttttctcgag caagaagggc ctcgacaagg ttatcaccgt gcaaaaaacc
     3121 gtcaccacca tcaagaccat cgttatcctg gattccaagg ttgactaccg cggttaccag
     3181 tgcctcgaca ccttcatcaa gcgtaacacc ccacccggct tccaagcctc ctccttcaaa
     3241 accgtggagg ttgatcgcaa ggagcaggtg gccctgatca tgaactcctc cggctccacc
     3301 ggtctgccaa agggcgtgca gctgacccac gagaacaccg tcacccgctt ctcgcatgcc
     3361 cgtgatccca tctacggcaa ccaggtgtcc ccaggcaccg ccgtcctgac cgtcgtgccc
     3421 ctccaccacg gcttcggcat gttcaccact ctcggctact tgatctgcgg cttccgtgtg
     3481 gtgatgctga ctaagttcga cgaggagact ttcctcaaga ccctgcaaga ctacaagtgc
     3541 acttacgtga tcctggtgcc gaccctgttc gccatcctga acaagagcga gctgttgaac
     3601 aaatacgacc tgtcgaacct ggtggagatc gcttccggcg gcgccccact gtcgaaggaa
     3661 gttggcgagg ccgttgctcg ccgcttcaac ctgcccggcg tccgtcaggg ttacggcctg
     3721 accgagacta cctcggctat catcatcacc cccgagggcg acgataagcc cggcgcttcc
     3781 ggcaaggttg ttccactctt caaagccaag gtgatcgacc tggacaccaa gaagtccctg
     3841 ggtcccaacc gtcgcggaga agtgtgcgtc aagggcccca tgctgatgaa gggctacgtc
     3901 aacaacccag aggccaccaa ggagttgatc gacgaggagg gctggctgca cactggtgac
     3961 atcggttact acgacgaaga gaagcacttc ttcatcgttg accgcctgaa gtccttgatc
     4021 aagtacaagg gctaccaggt tccccccgcc gagctcgaga gcgtcctgct gcagcacccc
     4081 tccattttcg atgccggagt ggccggtgtc cccgatcccg tggctggcga gctgccaggt
     4141 gccgtcgtcg tgctggagtc cggcaagaac atgaccgaga aggaagtgat ggattacgtg
     4201 gcctcgcaag tgtccaacgc caagcgcctg cgcggcggcg tgcgcttcgt ggatgaggtg
     4261 cccaagggcc tcaccggaaa gatcgacggt cgcgctatcc gcgagatcct caagaagccc
     4321 gtggccaaga tgtgagcttg gaatggatct tcgatcccga tcgttcaaac atttggcaat
     4381 aaagtttctt aagattgaat cctgttgccg gtcttgcgac gattatcata taatttctgt
     4441 tgaattacgt taagcatgta ataattaaca tgtaatgcat gacgttattt atgagatggg
     4501 tttttatgat tagagtcccg caattataca tttaatacgc gatagaaaac aaaatatagc
     4561 gcgcaaacta ggataaatta tcgcgcgcgg tgtcatctat gttactagat cgggaattgc
     4621 caagctaatt cttgaagatg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     4681 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     4741 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4801 tgataaatgc ttcaataatg ggaccgactc gcgctgtcat gagaccggat cctgacagga
     4861 tatattggcg ggtaaactaa gtcgctgtat gtgtttgttt gagatctcat gtgagcaaaa
     4921 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4981 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5041 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5101 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5161 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5221 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5281 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5341 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5401 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaagaaga
     5461 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5521 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5581 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5641 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5701 atatatgtgt aacattggtc tagtgattat ttgccgacta ccttggtgat ctcgcctttc
     5761 acgtagtgga caaattcttc caactgatct gcgcgcgagg ccaagcgatc ttcttcttgt
     5821 ccaagataag cctgtctagc ttcaagtatg acgggctgat actgggccgg caggcgctcc
     5881 attgcccagt cggcagcgac atccttcggc gcgattttgc cggttactgc gctgtaccaa
     5941 atgcgggaca acgtaagcac tacatttcgc tcatcaccag cccagtcggg cggcgagttc
     6001 catagcgtta aggtttcatt tagcgcctca aatagatcct gttcaggaac cggatcaaag
     6061 agttcctccg ccgctggacc taccaaggca acgctatgtt ctcttgcttt tgtcagcaag
     6121 atagccagat caatgtcgat cgtggctggc tcgaagatac ctgcaagaat gtcattgcgc
     6181 tgccattctc caaattgcag ttcgcgctta gctggataac gccacggaat gatgtcgtcg
     6241 tgcacaacaa tggtgacttc tacagcgcgg agaatctcgc tctctccagg ggaagccgaa
     6301 gtttccaaaa ggtcgttgat caaagctcgc cgcgttgttt catcaagcct tacggtcacc
     6361 gtaaccagca aatcaatatc actgtgtggc ttcaggccgc catccactgc ggagccgtac
     6421 aaatgtacgg ccagcaacgt cggttcgaga tggcgctcga tgacgccaac tacctctgat
     6481 agttgagtcg atacttcggc gatcaccgct tccctcataa tgtttaactt tgttttaggg
     6541 cgactgccct gctgcgtaac atcgttgctg ctccataaca tcaaacatcg acccacggcg
     6601 taacgcgctt gctgcttgga tgcccgaggc atagactgta ccccaaaaaa acagtcataa
     6661 caagccatga aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac
     6721 cagttgcgtg agcgcatacg ctacttgcat tacagcttac gaaccgaaca ggcttatgtc
     6781 cactgggttc gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac cttgggtagc
     6841 agcgaagtcg aggcatttct gtcctggctg gaacacccct tgtattactg tttatgtaag
     6901 cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat
     6961 tttgagacac aacgtggctt tgttgaataa atcgaacttt tgctgagttg aaggatcaga
     7021 tcacgcatct tcccgacaac gcagaccgtt ccgtggcaaa gcaaaagttc aaaatcacca
     7081 actggtccac ctacaacaaa gctctcatca accgtggctc cctcactttc tggctggatg
     7141 atggggcgat tcaggcgatc cccatccaac agcccgccgt cgagcgggct tttttatccc
     7201 cggaagcctg tggatagagg gtagttatcc acgtgaaacc gctaatgccc cgcaaagcct
     7261 tgattcacgg ggctttccgg cccgctccaa aaactatcca cgtgaaatcg ctaatcaggg
     7321 tacgtgaaat cgctaatcgg agtacgtgaa atcgctaata aggtcacgtg aaatcgctaa
     7381 tcaaaaaggc acgtgagaac gctaatagcc ctttcagatc aacagcttgc aaacacccct
     7441 cgctccggca agtagttaca gcaagtagta tgttcaatta gcttttcaat tatgaatata
     7501 tatatcaatt attggtcgcc cttggcttgt ggacaatgcg ctacgcgcac cggctccgcc
     7561 cgtggacaac cgcaagcggt tgcccaccgt cgagcgcctt tgcccacaac ccggcggccg
     7621 gccgcaacag atcgttttat aaattttttt ttttgaaaaa gaaaaagccc gaaaggcggc
     7681 aacctctcgg gcttctggat ttccgatccc cggaattaga tcttggcagg atatattgtg
     7741 gtg
//
