LOCUS       sequence-334479-        8136 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid Double Transcriptional Unit - 
            pAtUBQ10_E-Luc_NtEUt+pNOS_Red-F_NOSt.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8136)
  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 8136)
  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: pGREAT8
FEATURES             Location/Qualifiers
     source          1..8136
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             2131..2139
                     /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        2868..3047
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      4752..5004
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(5039..5057)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5246..5270
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5361..5949)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5441..5460)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6120..6911)
                     /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      7555..7990
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    join(8117..8136,1..3)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
ORIGIN
        1 taacggatcc ggtctcagga ggtcgacgag tcagtaataa acggcgtcaa agtggttgca
       61 gccggcacac acgagtcgtg tttatcaact caaagcacaa atacttttcc tcaacctaaa
      121 aataaggcaa ttagccaaaa acaactttgc gtgtaaacaa cgctcaatac acgtgtcatt
      181 ttattattag ctattgcttc accgccttag ctttctcgtg acctagtcgt cctcgtcttt
      241 tcttcttctt cttctataaa acaataccca aagagctctt cttcttcaca attcagattt
      301 caatttctca aaatcttaaa aactttctct caattctctc taccgtgatc aaggtaaatt
      361 tctgtgttcc ttattctctc aaaatcttcg attttgtttt cgttcgatcc caatttcgta
      421 tatgttcttt ggtttagatt ctgttaatct tagatcgaag atgattttct gggtttgatc
      481 gttagatatc atcttaattc tcgattaggg tttcatagat atcatccgat ttgttcaaat
      541 aatttgagtt ttgtcgaata attactcttc gatttgtgat ttctatctag atctggtgtt
      601 agtttctagt ttgtgcgatc gaatttgtcg attaatctga gtttttctga ttaacagaat
      661 ggagagagag aagaacgtgg tgtacggccc cgagcccaag caccctctgg gcaacttcac
      721 cgccggcgag atgctgtaca acgctctgca caagcactcc cacatccccc aggccatcct
      781 ggacgtgatg ggcaacgagt ccctttccta ccaggagttc ttcgacacta ctgtgaagct
      841 gggccagagc ctccagaact gtggctacaa gatgaacgat gtcgtgtcga tctgtgcaga
      901 gaacaacaag agattcttca tccccatcat ctccgcctgg tacatcggca tggtggtggc
      961 ccctgtgaac gaggactata tcccagacga gctgtgtaaa gtgaccggca tctccaagcc
     1021 gatcctggtc tttaccacta ggaagatcct gcctaaggtt ttggaggtta aagacagaac
     1081 caactacata aagaggatca tcatactgga ctctgaagag aacctgctgg gctgcgagag
     1141 cctgcacaac ttcatgtcca ggtactccga caacaacctc caaacattca agcctctgca
     1201 ctacgaccct gtggaccagg tagccgccat cctgtgctcc tccggcacaa ccggcctgcc
     1261 taaaggcgtg atgcagaccc acaggaacat ctgtgtgaga ctcacacacg catctgaccc
     1321 cagagtgggt acacaactca tccccggcgt atccgtgctg gcctacctgc cattcttcca
     1381 cgccttcggc ttcagtatca acctgggcta tttcatggtg ggcctgagag tggtgatgct
     1441 ccgaaggttt aaccaggagg tgttcctgaa ggccatccag gactacgagg tgaggagcgt
     1501 gatcaacgtt ccctccacaa tcctgttcct gtccaagagc cctctggtgg acaagtacga
     1561 cctatccacc ctggcggagc tgtgctgtgg agccgctcct ctggcgaagg aggtggccga
     1621 gatcgccgtg aagaggctga acctgccagg gatacggtgt ggctacggtc taacagagtc
     1681 tacctccgcc aacatccata ctctgcacaa cgagttcaag tccggctccc tgggcaaggt
     1741 gacaccttac atggccgcca agatcatcga caggaacacc ggcgaggccc tgggtccaaa
     1801 ccaggtgggc gagctgtgca tctggggacc tatggtaaca aaaggctatg tgaacaaccc
     1861 acaggctact aaggaggcca tcgacgacga cggctggctg cactctggcg acttcggcta
     1921 ctacgacgag gacgagtatt tctacatcgt ggaccggtac aaggagctga tcaaatacaa
     1981 gggctatcag gtcgcccctg tggagctgga ggagatcctc cttcagcacc caggcatcag
     2041 ggacgtggcc gtcgtgggta tccctgacat cgaggccggc gagctgccag ccggcttcgt
     2101 ggtgaagcag cccggcgccc aactcaccgc taaggaggtg tacgacttcc tggcccagag
     2161 ggtgtctcac tccaagtacc tgaggggcgg cgtaaggttc gtggactcta tccccaggaa
     2221 cgtgacaggc aagattagtc gaaaagagct gagggaggcc ctgatggaga aggcttctaa
     2281 gctgtaagct taaagcagaa tgctgagcta aaagaaaggc tttttccatt ttcgagagac
     2341 aatgagaaaa gaagaagaag aagaagaaga agaagaagaa gaagaagaaa agagtaaata
     2401 ataaagcccc acaggaggcg aagttcttgt agctccatgt tatctaagtt attgatattg
     2461 tttgccctat attttatttc tgtcattgtg tatgttttgt tcagtttcga tctccttgca
     2521 aaatgcagag attatgagat gaataaacta agttatatta ttatacgtgt taatattctc
     2581 ctcctctctc tagctagcct tttgttttct ctttttctta tttgattttc tttaaatcaa
     2641 tccattttag gagagggcca gggagtgatc cagcaaaaca tgaagattag aagaaacttc
     2701 cctctttttt ttcctgaaaa caatttaacg tcgagattta tctctttttg taatggaatc
     2761 atttctacag ttatgaccgc tgtcaggaga tacatgagaa ttaagggagt cacggtatga
     2821 cccccgccga tgacgcggga caagccgttt tacgtttgga actgacagaa ccgcaacgtt
     2881 gaaggagcca ctgagccgcg ggtttctgga gtttaatgag ctaagcacat acgtcagaaa
     2941 ccattattgc gcgttcaaaa gtcgcctaag gtcactatca gctagcaaat atttcttgtc
     3001 aaaaatgctc cactgacgtt ccataaattc ccctcggtat ccaattagag tctcatattc
     3061 actctcaact cgatcgaggc aatggagaac atggagaacg acgagaacat tgtggtggga
     3121 cccaagccgt tctacccgat cgaggagggt tccgctggca cccagctgcg caagtacatg
     3181 gagcgctacg ctaagctggg tgctatcgcc ttcaccaacg ccgtgaccgg tgtggattac
     3241 tcctacgccg agtacctgga gaagtcctgc tgcctcggaa aggccctgca gaactacggt
     3301 ctggtcgtgg atggccgcat tgccctctgc tcggagaact gcgaggagtt cttcattccc
     3361 gtgatcgctg gtctgttcat cggagtcggc gttgccccaa ccaacgagat ctacaccctg
     3421 cgcgagctgg tccacagcct gggtatcagc aagccgacca tcgttttctc gagcaagaag
     3481 ggcctcgaca aggttatcac cgtgcaaaaa accgtcacca ccatcaagac catcgttatc
     3541 ctggattcca aggttgacta ccgcggttac cagtgcctcg acaccttcat caagcgtaac
     3601 accccacccg gcttccaagc ctcctccttc aaaaccgtgg aggttgatcg caaggagcag
     3661 gtggccctga tcatgaactc ctccggctcc accggtctgc caaagggcgt gcagctgacc
     3721 cacgagaaca ccgtcacccg cttctcgcat gcccgtgatc ccatctacgg caaccaggtg
     3781 tccccaggca ccgccgtcct gaccgtcgtg cccctccacc acggcttcgg catgttcacc
     3841 actctcggct acttgatctg cggcttccgt gtggtgatgc tgactaagtt cgacgaggag
     3901 actttcctca agaccctgca agactacaag tgcacttacg tgatcctggt gccgaccctg
     3961 ttcgccatcc tgaacaagag cgagctgttg aacaaatacg acctgtcgaa cctggtggag
     4021 atcgcttccg gcggcgcccc actgtcgaag gaagttggcg aggccgttgc tcgccgcttc
     4081 aacctgcccg gcgtccgtca gggttacggc ctgaccgaga ctacctcggc tatcatcatc
     4141 acccccgagg gcgacgataa gcccggcgct tccggcaagg ttgttccact cttcaaagcc
     4201 aaggtgatcg acctggacac caagaagtcc ctgggtccca accgtcgcgg agaagtgtgc
     4261 gtcaagggcc ccatgctgat gaagggctac gtcaacaacc cagaggccac caaggagttg
     4321 atcgacgagg agggctggct gcacactggt gacatcggtt actacgacga agagaagcac
     4381 ttcttcatcg ttgaccgcct gaagtccttg atcaagtaca agggctacca ggttcccccc
     4441 gccgagctcg agagcgtcct gctgcagcac ccctccattt tcgatgccgg agtggccggt
     4501 gtccccgatc ccgtggctgg cgagctgcca ggtgccgtcg tcgtgctgga gtccggcaag
     4561 aacatgaccg agaaggaagt gatggattac gtggcctcgc aagtgtccaa cgccaagcgc
     4621 ctgcgcggcg gcgtgcgctt cgtggatgag gtgcccaagg gcctcaccgg aaagatcgac
     4681 ggtcgcgcta tccgcgagat cctcaagaag cccgtggcca agatgtgagc ttggaatgga
     4741 tcttcgatcc cgatcgttca aacatttggc aataaagttt cttaagattg aatcctgttg
     4801 ccggtcttgc gacgattatc atataatttc tgttgaatta cgttaagcat gtaataatta
     4861 acatgtaatg catgacgtta tttatgagat gggtttttat gattagagtc ccgcaattat
     4921 acatttaata cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg
     4981 cggtgtcatc tatgttacta gatcgggaat tgccaagcta attcttgaag atgaaagggc
     5041 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     5101 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     5161 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atgggaccga
     5221 ctcgcgctgt catgagaccg gatcctgaca ggatatattg gcgggtaaac taagtcgctg
     5281 tatgtgtttg tttgagatct catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5341 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5401 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5461 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5521 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5581 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5641 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5701 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5761 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5821 gctctgctga agccagttac cttcggaaga agagttggta gctcttgatc cggcaaacaa
     5881 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5941 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6001 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6061 aattaaaaat gaagttttaa atcaatctaa agtatatatg tgtaacattg gtctagtgat
     6121 tatttgccga ctaccttggt gatctcgcct ttcacgtagt ggacaaattc ttccaactga
     6181 tctgcgcgcg aggccaagcg atcttcttct tgtccaagat aagcctgtct agcttcaagt
     6241 atgacgggct gatactgggc cggcaggcgc tccattgccc agtcggcagc gacatccttc
     6301 ggcgcgattt tgccggttac tgcgctgtac caaatgcggg acaacgtaag cactacattt
     6361 cgctcatcac cagcccagtc gggcggcgag ttccatagcg ttaaggtttc atttagcgcc
     6421 tcaaatagat cctgttcagg aaccggatca aagagttcct ccgccgctgg acctaccaag
     6481 gcaacgctat gttctcttgc ttttgtcagc aagatagcca gatcaatgtc gatcgtggct
     6541 ggctcgaaga tacctgcaag aatgtcattg cgctgccatt ctccaaattg cagttcgcgc
     6601 ttagctggat aacgccacgg aatgatgtcg tcgtgcacaa caatggtgac ttctacagcg
     6661 cggagaatct cgctctctcc aggggaagcc gaagtttcca aaaggtcgtt gatcaaagct
     6721 cgccgcgttg tttcatcaag ccttacggtc accgtaacca gcaaatcaat atcactgtgt
     6781 ggcttcaggc cgccatccac tgcggagccg tacaaatgta cggccagcaa cgtcggttcg
     6841 agatggcgct cgatgacgcc aactacctct gatagttgag tcgatacttc ggcgatcacc
     6901 gcttccctca taatgtttaa ctttgtttta gggcgactgc cctgctgcgt aacatcgttg
     6961 ctgctccata acatcaaaca tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga
     7021 ggcatagact gtaccccaaa aaaacagtca taacaagcca tgaaaaccgc cactgcgccg
     7081 ttaccaccgc tgcgttcggt caaggttctg gaccagttgc gtgagcgcat acgctacttg
     7141 cattacagct tacgaaccga acaggcttat gtccactggg ttcgtgcctt catccgtttc
     7201 cacggtgtgc gtcacccggc aaccttgggt agcagcgaag tcgaggcatt tctgtcctgg
     7261 ctggaacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata
     7321 tatttttatc ttgtgcaatg taacatcaga gattttgaga cacaacgtgg ctttgttgaa
     7381 taaatcgaac ttttgctgag ttgaaggatc agatcacgca tcttcccgac aacgcagacc
     7441 gttccgtggc aaagcaaaag ttcaaaatca ccaactggtc cacctacaac aaagctctca
     7501 tcaaccgtgg ctccctcact ttctggctgg atgatggggc gattcaggcg atccccatcc
     7561 aacagcccgc cgtcgagcgg gcttttttat ccccggaagc ctgtggatag agggtagtta
     7621 tccacgtgaa accgctaatg ccccgcaaag ccttgattca cggggctttc cggcccgctc
     7681 caaaaactat ccacgtgaaa tcgctaatca gggtacgtga aatcgctaat cggagtacgt
     7741 gaaatcgcta ataaggtcac gtgaaatcgc taatcaaaaa ggcacgtgag aacgctaata
     7801 gccctttcag atcaacagct tgcaaacacc cctcgctccg gcaagtagtt acagcaagta
     7861 gtatgttcaa ttagcttttc aattatgaat atatatatca attattggtc gcccttggct
     7921 tgtggacaat gcgctacgcg caccggctcc gcccgtggac aaccgcaagc ggttgcccac
     7981 cgtcgagcgc ctttgcccac aacccggcgg ccggccgcaa cagatcgttt tataaatttt
     8041 tttttttgaa aaagaaaaag cccgaaaggc ggcaacctct cgggcttctg gatttccgat
     8101 ccccggaatt agatcttggc aggatatatt gtggtg
//
