LOCUS       sequence-341132-        7963 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid Double Transcriptional Unit - 
            pNOS_E-Luc_AtACT2t+pNOS_Red-F_NOSt.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7963)
  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 7963)
  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: pGREAT11
FEATURES             Location/Qualifiers
     source          1..7963
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(108..287)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    384..406
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     rep_origin      533..968
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     CDS             1612..2403
                     /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      2574..3162
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3063..3082
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    3253..3277
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     3466..3484
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3519..3771)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(5476..5655)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     RBS             6556..6564
                     /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)"
ORIGIN
        1 tctcgccggc ggtgaagttg cccagagggt gcttgggctc ggggccgtac accacgttct
       61 tctctctctc cattgcctcg atcgagttga gagtgaatat gagactctaa ttggataccg
      121 aggggaattt atggaacgtc agtggagcat ttttgacaag aaatatttgc tagctgatag
      181 tgaccttagg cgacttttga acgcgcaata atggtttctg acgtatgtgc ttagctcatt
      241 aaactccaga aacccgcggc tcagtggctc cttcaacgtt gcggttctgt cagttccaaa
      301 cgtaaaacgg cttgtcccgc gtcatcggcg ggggtcatac cgtgactccc ttaattctca
      361 tgtatctcct gagaccggat ccgttacacc acaatatatc ctgccaagat ctaattccgg
      421 ggatcggaaa tccagaagcc cgagaggttg ccgcctttcg ggctttttct ttttcaaaaa
      481 aaaaaattta taaaacgatc tgttgcggcc ggccgccggg ttgtgggcaa aggcgctcga
      541 cggtgggcaa ccgcttgcgg ttgtccacgg gcggagccgg tgcgcgtagc gcattgtcca
      601 caagccaagg gcgaccaata attgatatat atattcataa ttgaaaagct aattgaacat
      661 actacttgct gtaactactt gccggagcga ggggtgtttg caagctgttg atctgaaagg
      721 gctattagcg ttctcacgtg cctttttgat tagcgatttc acgtgacctt attagcgatt
      781 tcacgtactc cgattagcga tttcacgtac cctgattagc gatttcacgt ggatagtttt
      841 tggagcgggc cggaaagccc cgtgaatcaa ggctttgcgg ggcattagcg gtttcacgtg
      901 gataactacc ctctatccac aggcttccgg ggataaaaaa gcccgctcga cggcgggctg
      961 ttggatgggg atcgcctgaa tcgccccatc atccagccag aaagtgaggg agccacggtt
     1021 gatgagagct ttgttgtagg tggaccagtt ggtgattttg aacttttgct ttgccacgga
     1081 acggtctgcg ttgtcgggaa gatgcgtgat ctgatccttc aactcagcaa aagttcgatt
     1141 tattcaacaa agccacgttg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa
     1201 tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttcc
     1261 agccaggaca gaaatgcctc gacttcgctg ctacccaagg ttgccgggtg acgcacaccg
     1321 tggaaacgga tgaaggcacg aacccagtgg acataagcct gttcggttcg taagctgtaa
     1381 tgcaagtagc gtatgcgctc acgcaactgg tccagaacct tgaccgaacg cagcggtggt
     1441 aacggcgcag tggcggtttt catggcttgt tatgactgtt tttttggggt acagtctatg
     1501 cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc
     1561 agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa agttaaacat tatgagggaa
     1621 gcggtgatcg ccgaagtatc gactcaacta tcagaggtag ttggcgtcat cgagcgccat
     1681 ctcgaaccga cgttgctggc cgtacatttg tacggctccg cagtggatgg cggcctgaag
     1741 ccacacagtg atattgattt gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg
     1801 cgagctttga tcaacgacct tttggaaact tcggcttccc ctggagagag cgagattctc
     1861 cgcgctgtag aagtcaccat tgttgtgcac gacgacatca ttccgtggcg ttatccagct
     1921 aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg tatcttcgag
     1981 ccagccacga tcgacattga tctggctatc ttgctgacaa aagcaagaga acatagcgtt
     2041 gccttggtag gtccagcggc ggaggaactc tttgatccgg ttcctgaaca ggatctattt
     2101 gaggcgctaa atgaaacctt aacgctatgg aactcgccgc ccgactgggc tggtgatgag
     2161 cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg
     2221 ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc
     2281 atacttgaag ctagacaggc ttatcttgga caagaagaag atcgcttggc ctcgcgcgca
     2341 gatcagttgg aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa
     2401 taatcactag accaatgtta cacatatata ctttagattg atttaaaact tcatttttaa
     2461 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     2521 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2581 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2641 gtttgtttgc cggatcaaga gctaccaact cttcttccga aggtaactgg cttcagcaga
     2701 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     2761 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2821 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2881 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2941 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3001 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3061 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3121 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3181 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgagatctca aacaaacaca
     3241 tacagcgact tagtttaccc gccaatatat cctgtcagga tccggtctca tgacagcgcg
     3301 agtcggtccc attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     3361 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     3421 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     3481 aggccctttc atcttcaaga attagcttgg caattcccga tctagtaaca tagatgacac
     3541 cgcgcgcgat aatttatcct agtttgcgcg ctatattttg ttttctatcg cgtattaaat
     3601 gtataattgc gggactctaa tcataaaaac ccatctcata aataacgtca tgcattacat
     3661 gttaattatt acatgcttaa cgtaattcaa cagaaattat atgataatcg tcgcaagacc
     3721 ggcaacagga ttcaatctta agaaacttta ttgccaaatg tttgaacgat cgggatcgaa
     3781 gatccattcc aagctcacat cttggccacg ggcttcttga ggatctcgcg gatagcgcga
     3841 ccgtcgatct ttccggtgag gcccttgggc acctcatcca cgaagcgcac gccgccgcgc
     3901 aggcgcttgg cgttggacac ttgcgaggcc acgtaatcca tcacttcctt ctcggtcatg
     3961 ttcttgccgg actccagcac gacgacggca cctggcagct cgccagccac gggatcgggg
     4021 acaccggcca ctccggcatc gaaaatggag gggtgctgca gcaggacgct ctcgagctcg
     4081 gcggggggaa cctggtagcc cttgtacttg atcaaggact tcaggcggtc aacgatgaag
     4141 aagtgcttct cttcgtcgta gtaaccgatg tcaccagtgt gcagccagcc ctcctcgtcg
     4201 atcaactcct tggtggcctc tgggttgttg acgtagccct tcatcagcat ggggcccttg
     4261 acgcacactt ctccgcgacg gttgggaccc agggacttct tggtgtccag gtcgatcacc
     4321 ttggctttga agagtggaac aaccttgccg gaagcgccgg gcttatcgtc gccctcgggg
     4381 gtgatgatga tagccgaggt agtctcggtc aggccgtaac cctgacggac gccgggcagg
     4441 ttgaagcggc gagcaacggc ctcgccaact tccttcgaca gtggggcgcc gccggaagcg
     4501 atctccacca ggttcgacag gtcgtatttg ttcaacagct cgctcttgtt caggatggcg
     4561 aacagggtcg gcaccaggat cacgtaagtg cacttgtagt cttgcagggt cttgaggaaa
     4621 gtctcctcgt cgaacttagt cagcatcacc acacggaagc cgcagatcaa gtagccgaga
     4681 gtggtgaaca tgccgaagcc gtggtggagg ggcacgacgg tcaggacggc ggtgcctggg
     4741 gacacctggt tgccgtagat gggatcacgg gcatgcgaga agcgggtgac ggtgttctcg
     4801 tgggtcagct gcacgccctt tggcagaccg gtggagccgg aggagttcat gatcagggcc
     4861 acctgctcct tgcgatcaac ctccacggtt ttgaaggagg aggcttggaa gccgggtggg
     4921 gtgttacgct tgatgaaggt gtcgaggcac tggtaaccgc ggtagtcaac cttggaatcc
     4981 aggataacga tggtcttgat ggtggtgacg gttttttgca cggtgataac cttgtcgagg
     5041 cccttcttgc tcgagaaaac gatggtcggc ttgctgatac ccaggctgtg gaccagctcg
     5101 cgcagggtgt agatctcgtt ggttggggca acgccgactc cgatgaacag accagcgatc
     5161 acgggaatga agaactcctc gcagttctcc gagcagaggg caatgcggcc atccacgacc
     5221 agaccgtagt tctgcagggc ctttccgagg cagcaggact tctccaggta ctcggcgtag
     5281 gagtaatcca caccggtcac ggcgttggtg aaggcgatag cacccagctt agcgtagcgc
     5341 tccatgtact tgcgcagctg ggtgccagcg gaaccctcct cgatcgggta gaacggcttg
     5401 ggtcccacca caatgttctc gtcgttctcc atgttctcca ttgcctcgat cgagttgaga
     5461 gtgaatatga gactctaatt ggataccgag gggaatttat ggaacgtcag tggagcattt
     5521 ttgacaagaa atatttgcta gctgatagtg accttaggcg acttttgaac gcgcaataat
     5581 ggtttctgac gtatgtgctt agctcattaa actccagaaa cccgcggctc agtggctcct
     5641 tcaacgttgc ggttctgtca gttccaaacg taaaacggct tgtcccgcgt catcggcggg
     5701 ggtcataccg tgactccctt aattctcatg tatctcctga cagcgacttt ttggaatcaa
     5761 gatgtataaa ccaaatgtgc aggacaattt gaaattaatt tgttatattt tgcatcatgt
     5821 aaaaggatta ataagcgaaa agagtaacga atagatacac ctcgtaattg ttttggtgta
     5881 aaacttgaaa cagtagaatt gacgattggg gttatataaa atggtgcgat cttaaaatat
     5941 ctgactttgt gctagtttaa gttcatttgt gaatggagga catgtaacgc tttaactatt
     6001 cagaatatca gttccaagtg aagatggagg tagcaaaagt aagactaata accaaatcaa
     6061 taaaaatctt atacaatact tatattaaca ttgcaaagag tttcaaggtt tttcttctga
     6121 gttttgaaag aaattatagg gaacaaaagg aataaagagg catcaattcg atcactcaga
     6181 gctacaaaac aatgggacta aaacgcaaaa cgaaagcggt tttttttttt gaaacgcaga
     6241 cgtaagtaaa aacccagaga gtttgtcaca cacaagtgca tcatagaaac gaaaacaaaa
     6301 agggaaatga aacaaacaaa tggagaagca aatataaaag aaaaaaagaa actttgatcc
     6361 cattcataaa accccagctt tttaagcctt tgatcttgag agcaagctta cagcttagaa
     6421 gccttctcca tcagggcctc cctcagctct tttcgactaa tcttgcctgt cacgttcctg
     6481 gggatagagt ccacgaacct tacgccgccc ctcaggtact tggagtgaga caccctctgg
     6541 gccaggaagt cgtacacctc cttagcggtg agttgggcgc cgggctgctt caccacgaag
     6601 ccggctggca gctcgccggc ctcgatgtca gggataccca cgacggccac gtccctgatg
     6661 cctgggtgct gaaggaggat ctcctccagc tccacagggg cgacctgata gcccttgtat
     6721 ttgatcagct ccttgtaccg gtccacgatg tagaaatact cgtcctcgtc gtagtagccg
     6781 aagtcgccag agtgcagcca gccgtcgtcg tcgatggcct ccttagtagc ctgtgggttg
     6841 ttcacatagc cttttgttac cataggtccc cagatgcaca gctcgcccac ctggtttgga
     6901 cccagggcct cgccggtgtt cctgtcgatg atcttggcgg ccatgtaagg tgtcaccttg
     6961 cccagggagc cggacttgaa ctcgttgtgc agagtatgga tgttggcgga ggtagactct
     7021 gttagaccgt agccacaccg tatccctggc aggttcagcc tcttcacggc gatctcggcc
     7081 acctccttcg ccagaggagc ggctccacag cacagctccg ccagggtgga taggtcgtac
     7141 ttgtccacca gagggctctt ggacaggaac aggattgtgg agggaacgtt gatcacgctc
     7201 ctcacctcgt agtcctggat ggccttcagg aacacctcct ggttaaacct tcggagcatc
     7261 accactctca ggcccaccat gaaatagccc aggttgatac tgaagccgaa ggcgtggaag
     7321 aatggcaggt aggccagcac ggatacgccg gggatgagtt gtgtacccac tctggggtca
     7381 gatgcgtgtg tgagtctcac acagatgttc ctgtgggtct gcatcacgcc tttaggcagg
     7441 ccggttgtgc cggaggagca caggatggcg gctacctggt ccacagggtc gtagtgcaga
     7501 ggcttgaatg tttggaggtt gttgtcggag tacctggaca tgaagttgtg caggctctcg
     7561 cagcccagca ggttctcttc agagtccagt atgatgatcc tctttatgta gttggttctg
     7621 tctttaacct ccaaaacctt aggcaggatc ttcctagtgg taaagaccag gatcggcttg
     7681 gagatgccgg tcactttaca cagctcgtct gggatatagt cctcgttcac aggggccacc
     7741 accatgccga tgtaccaggc ggagatgatg gggatgaaga atctcttgtt gttctctgca
     7801 cagatcgaca cgacatcgtt catcttgtag ccacagttct ggaggctctg gcccagcttc
     7861 acagtagtgt cgaagaactc ctggtaggaa agggactcgt tgcccatcac gtccaggatg
     7921 gcctggggga tgtgggagtg cttgtgcaga gcgttgtaca gca
//
