LOCUS       sequence-334497-        7483 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid Double Transcriptional Unit - 
            pG10-90_E-Luc_35St+pNOS_Red-F_NOSt.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7483)
  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 7483)
  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: pGREAT24
FEATURES             Location/Qualifiers
     source          1..7483
                     /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        2215..2394
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      4099..4351
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4386..4404)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4593..4617
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4708..5296)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4788..4807)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5467..6258)
                     /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      6902..7337
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    join(7464..7483,1..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 gtaagcttcg gccatgctag agtccgcaaa
     1921 aatcaccagt ctctctctac aaatctatct ctctctattt ttctccagaa taatgtgtga
     1981 gtagttccca gataagggaa ttagggttct tatagggttt cgctcatgtg ttgagcatat
     2041 aagaaaccct tagtatgtat ttgtatttgt aaaatacttc tatcaataaa atttctaatt
     2101 cctaaaacca aaatccagtg acctcgctgt caggagatac atgagaatta agggagtcac
     2161 ggtatgaccc ccgccgatga cgcgggacaa gccgttttac gtttggaact gacagaaccg
     2221 caacgttgaa ggagccactg agccgcgggt ttctggagtt taatgagcta agcacatacg
     2281 tcagaaacca ttattgcgcg ttcaaaagtc gcctaaggtc actatcagct agcaaatatt
     2341 tcttgtcaaa aatgctccac tgacgttcca taaattcccc tcggtatcca attagagtct
     2401 catattcact ctcaactcga tcgaggcaat ggagaacatg gagaacgacg agaacattgt
     2461 ggtgggaccc aagccgttct acccgatcga ggagggttcc gctggcaccc agctgcgcaa
     2521 gtacatggag cgctacgcta agctgggtgc tatcgccttc accaacgccg tgaccggtgt
     2581 ggattactcc tacgccgagt acctggagaa gtcctgctgc ctcggaaagg ccctgcagaa
     2641 ctacggtctg gtcgtggatg gccgcattgc cctctgctcg gagaactgcg aggagttctt
     2701 cattcccgtg atcgctggtc tgttcatcgg agtcggcgtt gccccaacca acgagatcta
     2761 caccctgcgc gagctggtcc acagcctggg tatcagcaag ccgaccatcg ttttctcgag
     2821 caagaagggc ctcgacaagg ttatcaccgt gcaaaaaacc gtcaccacca tcaagaccat
     2881 cgttatcctg gattccaagg ttgactaccg cggttaccag tgcctcgaca ccttcatcaa
     2941 gcgtaacacc ccacccggct tccaagcctc ctccttcaaa accgtggagg ttgatcgcaa
     3001 ggagcaggtg gccctgatca tgaactcctc cggctccacc ggtctgccaa agggcgtgca
     3061 gctgacccac gagaacaccg tcacccgctt ctcgcatgcc cgtgatccca tctacggcaa
     3121 ccaggtgtcc ccaggcaccg ccgtcctgac cgtcgtgccc ctccaccacg gcttcggcat
     3181 gttcaccact ctcggctact tgatctgcgg cttccgtgtg gtgatgctga ctaagttcga
     3241 cgaggagact ttcctcaaga ccctgcaaga ctacaagtgc acttacgtga tcctggtgcc
     3301 gaccctgttc gccatcctga acaagagcga gctgttgaac aaatacgacc tgtcgaacct
     3361 ggtggagatc gcttccggcg gcgccccact gtcgaaggaa gttggcgagg ccgttgctcg
     3421 ccgcttcaac ctgcccggcg tccgtcaggg ttacggcctg accgagacta cctcggctat
     3481 catcatcacc cccgagggcg acgataagcc cggcgcttcc ggcaaggttg ttccactctt
     3541 caaagccaag gtgatcgacc tggacaccaa gaagtccctg ggtcccaacc gtcgcggaga
     3601 agtgtgcgtc aagggcccca tgctgatgaa gggctacgtc aacaacccag aggccaccaa
     3661 ggagttgatc gacgaggagg gctggctgca cactggtgac atcggttact acgacgaaga
     3721 gaagcacttc ttcatcgttg accgcctgaa gtccttgatc aagtacaagg gctaccaggt
     3781 tccccccgcc gagctcgaga gcgtcctgct gcagcacccc tccattttcg atgccggagt
     3841 ggccggtgtc cccgatcccg tggctggcga gctgccaggt gccgtcgtcg tgctggagtc
     3901 cggcaagaac atgaccgaga aggaagtgat ggattacgtg gcctcgcaag tgtccaacgc
     3961 caagcgcctg cgcggcggcg tgcgcttcgt ggatgaggtg cccaagggcc tcaccggaaa
     4021 gatcgacggt cgcgctatcc gcgagatcct caagaagccc gtggccaaga tgtgagcttg
     4081 gaatggatct tcgatcccga tcgttcaaac atttggcaat aaagtttctt aagattgaat
     4141 cctgttgccg gtcttgcgac gattatcata taatttctgt tgaattacgt taagcatgta
     4201 ataattaaca tgtaatgcat gacgttattt atgagatggg tttttatgat tagagtcccg
     4261 caattataca tttaatacgc gatagaaaac aaaatatagc gcgcaaacta ggataaatta
     4321 tcgcgcgcgg tgtcatctat gttactagat cgggaattgc caagctaatt cttgaagatg
     4381 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     4441 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4501 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataatg
     4561 ggaccgactc gcgctgtcat gagaccggat cctgacagga tatattggcg ggtaaactaa
     4621 gtcgctgtat gtgtttgttt gagatctcat gtgagcaaaa ggccagcaaa aggccaggaa
     4681 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4741 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4801 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4861 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4921 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4981 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5041 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5101 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     5161 tatctgcgct ctgctgaagc cagttacctt cggaagaaga gttggtagct cttgatccgg
     5221 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5281 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5341 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5401 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgtgt aacattggtc
     5461 tagtgattat ttgccgacta ccttggtgat ctcgcctttc acgtagtgga caaattcttc
     5521 caactgatct gcgcgcgagg ccaagcgatc ttcttcttgt ccaagataag cctgtctagc
     5581 ttcaagtatg acgggctgat actgggccgg caggcgctcc attgcccagt cggcagcgac
     5641 atccttcggc gcgattttgc cggttactgc gctgtaccaa atgcgggaca acgtaagcac
     5701 tacatttcgc tcatcaccag cccagtcggg cggcgagttc catagcgtta aggtttcatt
     5761 tagcgcctca aatagatcct gttcaggaac cggatcaaag agttcctccg ccgctggacc
     5821 taccaaggca acgctatgtt ctcttgcttt tgtcagcaag atagccagat caatgtcgat
     5881 cgtggctggc tcgaagatac ctgcaagaat gtcattgcgc tgccattctc caaattgcag
     5941 ttcgcgctta gctggataac gccacggaat gatgtcgtcg tgcacaacaa tggtgacttc
     6001 tacagcgcgg agaatctcgc tctctccagg ggaagccgaa gtttccaaaa ggtcgttgat
     6061 caaagctcgc cgcgttgttt catcaagcct tacggtcacc gtaaccagca aatcaatatc
     6121 actgtgtggc ttcaggccgc catccactgc ggagccgtac aaatgtacgg ccagcaacgt
     6181 cggttcgaga tggcgctcga tgacgccaac tacctctgat agttgagtcg atacttcggc
     6241 gatcaccgct tccctcataa tgtttaactt tgttttaggg cgactgccct gctgcgtaac
     6301 atcgttgctg ctccataaca tcaaacatcg acccacggcg taacgcgctt gctgcttgga
     6361 tgcccgaggc atagactgta ccccaaaaaa acagtcataa caagccatga aaaccgccac
     6421 tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg agcgcatacg
     6481 ctacttgcat tacagcttac gaaccgaaca ggcttatgtc cactgggttc gtgccttcat
     6541 ccgtttccac ggtgtgcgtc acccggcaac cttgggtagc agcgaagtcg aggcatttct
     6601 gtcctggctg gaacacccct tgtattactg tttatgtaag cagacagttt tattgttcat
     6661 gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac aacgtggctt
     6721 tgttgaataa atcgaacttt tgctgagttg aaggatcaga tcacgcatct tcccgacaac
     6781 gcagaccgtt ccgtggcaaa gcaaaagttc aaaatcacca actggtccac ctacaacaaa
     6841 gctctcatca accgtggctc cctcactttc tggctggatg atggggcgat tcaggcgatc
     6901 cccatccaac agcccgccgt cgagcgggct tttttatccc cggaagcctg tggatagagg
     6961 gtagttatcc acgtgaaacc gctaatgccc cgcaaagcct tgattcacgg ggctttccgg
     7021 cccgctccaa aaactatcca cgtgaaatcg ctaatcaggg tacgtgaaat cgctaatcgg
     7081 agtacgtgaa atcgctaata aggtcacgtg aaatcgctaa tcaaaaaggc acgtgagaac
     7141 gctaatagcc ctttcagatc aacagcttgc aaacacccct cgctccggca agtagttaca
     7201 gcaagtagta tgttcaatta gcttttcaat tatgaatata tatatcaatt attggtcgcc
     7261 cttggcttgt ggacaatgcg ctacgcgcac cggctccgcc cgtggacaac cgcaagcggt
     7321 tgcccaccgt cgagcgcctt tgcccacaac ccggcggccg gccgcaacag atcgttttat
     7381 aaattttttt ttttgaaaaa gaaaaagccc gaaaggcggc aacctctcgg gcttctggat
     7441 ttccgatccc cggaattaga tcttggcagg atatattgtg gtg
//
