LOCUS       sequence-334469-        7749 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid Double Transcriptional Unit - 
            pG10-90_E-Luc_NOSt+pNOS_Red-F_NOSt.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7749)
  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 7749)
  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: pGREAT4
FEATURES             Location/Qualifiers
     source          1..7749
                     /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)"
     terminator      1918..2170
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        2481..2660
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      4365..4617
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    4859..4883
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4974..5562)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5054..5073)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5733..6524)
                     /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      7168..7603
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    join(7730..7749,1..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 gtaagcttgg aatggatctt cgatcccgat
     1921 cgttcaaaca tttggcaata aagtttctta agattgaatc ctgttgccgg tcttgcgacg
     1981 attatcatat aatttctgtt gaattacgtt aagcatgtaa taattaacat gtaatgcatg
     2041 acgttattta tgagatgggt ttttatgatt agagtcccgc aattatacat ttaatacgcg
     2101 atagaaaaca aaatatagcg cgcaaactag gataaattat cgcgcgcggt gtcatctatg
     2161 ttactagatc gggaattgcc aagctaattc ttgaagatga aagggcctcg tgatacgcct
     2221 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     2281 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2341 gctcatgaga caataaccct gataaatgct tcaataatgg gaccgactcg cgctgtcagg
     2401 agatacatga gaattaaggg agtcacggta tgacccccgc cgatgacgcg ggacaagccg
     2461 ttttacgttt ggaactgaca gaaccgcaac gttgaaggag ccactgagcc gcgggtttct
     2521 ggagtttaat gagctaagca catacgtcag aaaccattat tgcgcgttca aaagtcgcct
     2581 aaggtcacta tcagctagca aatatttctt gtcaaaaatg ctccactgac gttccataaa
     2641 ttcccctcgg tatccaatta gagtctcata ttcactctca actcgatcga ggcaatggag
     2701 aacatggaga acgacgagaa cattgtggtg ggacccaagc cgttctaccc gatcgaggag
     2761 ggttccgctg gcacccagct gcgcaagtac atggagcgct acgctaagct gggtgctatc
     2821 gccttcacca acgccgtgac cggtgtggat tactcctacg ccgagtacct ggagaagtcc
     2881 tgctgcctcg gaaaggccct gcagaactac ggtctggtcg tggatggccg cattgccctc
     2941 tgctcggaga actgcgagga gttcttcatt cccgtgatcg ctggtctgtt catcggagtc
     3001 ggcgttgccc caaccaacga gatctacacc ctgcgcgagc tggtccacag cctgggtatc
     3061 agcaagccga ccatcgtttt ctcgagcaag aagggcctcg acaaggttat caccgtgcaa
     3121 aaaaccgtca ccaccatcaa gaccatcgtt atcctggatt ccaaggttga ctaccgcggt
     3181 taccagtgcc tcgacacctt catcaagcgt aacaccccac ccggcttcca agcctcctcc
     3241 ttcaaaaccg tggaggttga tcgcaaggag caggtggccc tgatcatgaa ctcctccggc
     3301 tccaccggtc tgccaaaggg cgtgcagctg acccacgaga acaccgtcac ccgcttctcg
     3361 catgcccgtg atcccatcta cggcaaccag gtgtccccag gcaccgccgt cctgaccgtc
     3421 gtgcccctcc accacggctt cggcatgttc accactctcg gctacttgat ctgcggcttc
     3481 cgtgtggtga tgctgactaa gttcgacgag gagactttcc tcaagaccct gcaagactac
     3541 aagtgcactt acgtgatcct ggtgccgacc ctgttcgcca tcctgaacaa gagcgagctg
     3601 ttgaacaaat acgacctgtc gaacctggtg gagatcgctt ccggcggcgc cccactgtcg
     3661 aaggaagttg gcgaggccgt tgctcgccgc ttcaacctgc ccggcgtccg tcagggttac
     3721 ggcctgaccg agactacctc ggctatcatc atcacccccg agggcgacga taagcccggc
     3781 gcttccggca aggttgttcc actcttcaaa gccaaggtga tcgacctgga caccaagaag
     3841 tccctgggtc ccaaccgtcg cggagaagtg tgcgtcaagg gccccatgct gatgaagggc
     3901 tacgtcaaca acccagaggc caccaaggag ttgatcgacg aggagggctg gctgcacact
     3961 ggtgacatcg gttactacga cgaagagaag cacttcttca tcgttgaccg cctgaagtcc
     4021 ttgatcaagt acaagggcta ccaggttccc cccgccgagc tcgagagcgt cctgctgcag
     4081 cacccctcca ttttcgatgc cggagtggcc ggtgtccccg atcccgtggc tggcgagctg
     4141 ccaggtgccg tcgtcgtgct ggagtccggc aagaacatga ccgagaagga agtgatggat
     4201 tacgtggcct cgcaagtgtc caacgccaag cgcctgcgcg gcggcgtgcg cttcgtggat
     4261 gaggtgccca agggcctcac cggaaagatc gacggtcgcg ctatccgcga gatcctcaag
     4321 aagcccgtgg ccaagatgtg agcttggaat ggatcttcga tcccgatcgt tcaaacattt
     4381 ggcaataaag tttcttaaga ttgaatcctg ttgccggtct tgcgacgatt atcatataat
     4441 ttctgttgaa ttacgttaag catgtaataa ttaacatgta atgcatgacg ttatttatga
     4501 gatgggtttt tatgattaga gtcccgcaat tatacattta atacgcgata gaaaacaaaa
     4561 tatagcgcgc aaactaggat aaattatcgc gcgcggtgtc atctatgtta ctagatcggg
     4621 aattgccaag ctaattcttg aagatgaaag ggcctcgtga tacgcctatt tttataggtt
     4681 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     4741 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4801 taaccctgat aaatgcttca ataatgggac cgactcgcgc tgtcatgaga ccggatcctg
     4861 acaggatata ttggcgggta aactaagtcg ctgtatgtgt ttgtttgaga tctcatgtga
     4921 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4981 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5041 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5101 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5161 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5221 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5281 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5341 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5401 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5461 agaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5521 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5581 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5641 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5701 taaagtatat atgtgtaaca ttggtctagt gattatttgc cgactacctt ggtgatctcg
     5761 cctttcacgt agtggacaaa ttcttccaac tgatctgcgc gcgaggccaa gcgatcttct
     5821 tcttgtccaa gataagcctg tctagcttca agtatgacgg gctgatactg ggccggcagg
     5881 cgctccattg cccagtcggc agcgacatcc ttcggcgcga ttttgccggt tactgcgctg
     5941 taccaaatgc gggacaacgt aagcactaca tttcgctcat caccagccca gtcgggcggc
     6001 gagttccata gcgttaaggt ttcatttagc gcctcaaata gatcctgttc aggaaccgga
     6061 tcaaagagtt cctccgccgc tggacctacc aaggcaacgc tatgttctct tgcttttgtc
     6121 agcaagatag ccagatcaat gtcgatcgtg gctggctcga agatacctgc aagaatgtca
     6181 ttgcgctgcc attctccaaa ttgcagttcg cgcttagctg gataacgcca cggaatgatg
     6241 tcgtcgtgca caacaatggt gacttctaca gcgcggagaa tctcgctctc tccaggggaa
     6301 gccgaagttt ccaaaaggtc gttgatcaaa gctcgccgcg ttgtttcatc aagccttacg
     6361 gtcaccgtaa ccagcaaatc aatatcactg tgtggcttca ggccgccatc cactgcggag
     6421 ccgtacaaat gtacggccag caacgtcggt tcgagatggc gctcgatgac gccaactacc
     6481 tctgatagtt gagtcgatac ttcggcgatc accgcttccc tcataatgtt taactttgtt
     6541 ttagggcgac tgccctgctg cgtaacatcg ttgctgctcc ataacatcaa acatcgaccc
     6601 acggcgtaac gcgcttgctg cttggatgcc cgaggcatag actgtacccc aaaaaaacag
     6661 tcataacaag ccatgaaaac cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt
     6721 ctggaccagt tgcgtgagcg catacgctac ttgcattaca gcttacgaac cgaacaggct
     6781 tatgtccact gggttcgtgc cttcatccgt ttccacggtg tgcgtcaccc ggcaaccttg
     6841 ggtagcagcg aagtcgaggc atttctgtcc tggctggaac accccttgta ttactgttta
     6901 tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca atgtaacatc
     6961 agagattttg agacacaacg tggctttgtt gaataaatcg aacttttgct gagttgaagg
     7021 atcagatcac gcatcttccc gacaacgcag accgttccgt ggcaaagcaa aagttcaaaa
     7081 tcaccaactg gtccacctac aacaaagctc tcatcaaccg tggctccctc actttctggc
     7141 tggatgatgg ggcgattcag gcgatcccca tccaacagcc cgccgtcgag cgggcttttt
     7201 tatccccgga agcctgtgga tagagggtag ttatccacgt gaaaccgcta atgccccgca
     7261 aagccttgat tcacggggct ttccggcccg ctccaaaaac tatccacgtg aaatcgctaa
     7321 tcagggtacg tgaaatcgct aatcggagta cgtgaaatcg ctaataaggt cacgtgaaat
     7381 cgctaatcaa aaaggcacgt gagaacgcta atagcccttt cagatcaaca gcttgcaaac
     7441 acccctcgct ccggcaagta gttacagcaa gtagtatgtt caattagctt ttcaattatg
     7501 aatatatata tcaattattg gtcgcccttg gcttgtggac aatgcgctac gcgcaccggc
     7561 tccgcccgtg gacaaccgca agcggttgcc caccgtcgag cgcctttgcc cacaacccgg
     7621 cggccggccg caacagatcg ttttataaat tttttttttt gaaaaagaaa aagcccgaaa
     7681 ggcggcaacc tctcgggctt ctggatttcc gatccccgga attagatctt ggcaggatat
     7741 attgtggtg
//
