LOCUS       sequence-334553-        8308 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid Double Transcriptional Unit - 
            pAtUBQ10_E-Luc_AtACT2t+pNOS_Red-F_NOSt.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8308)
  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 8308)
  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: pGREAT13
FEATURES             Location/Qualifiers
     source          1..8308
                     /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        3040..3219
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      4924..5176
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(5211..5229)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5418..5442
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5533..6121)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5613..5632)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6292..7083)
                     /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      7727..8162
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    join(8289..8308,1..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 tgctctcaag atcaaaggct taaaaagctg gggttttatg aatgggatca
     2341 aagtttcttt ttttctttta tatttgcttc tccatttgtt tgtttcattt ccctttttgt
     2401 tttcgtttct atgatgcact tgtgtgtgac aaactctctg ggtttttact tacgtctgcg
     2461 tttcaaaaaa aaaaaccgct ttcgttttgc gttttagtcc cattgttttg tagctctgag
     2521 tgatcgaatt gatgcctctt tattcctttt gttccctata atttctttca aaactcagaa
     2581 gaaaaacctt gaaactcttt gcaatgttaa tataagtatt gtataagatt tttattgatt
     2641 tggttattag tcttactttt gctacctcca tcttcacttg gaactgatat tctgaatagt
     2701 taaagcgtta catgtcctcc attcacaaat gaacttaaac tagcacaaag tcagatattt
     2761 taagatcgca ccattttata taaccccaat cgtcaattct actgtttcaa gttttacacc
     2821 aaaacaatta cgaggtgtat ctattcgtta ctcttttcgc ttattaatcc ttttacatga
     2881 tgcaaaatat aacaaattaa tttcaaattg tcctgcacat ttggtttata catcttgatt
     2941 ccaaaaagtc gctgtcagga gatacatgag aattaaggga gtcacggtat gacccccgcc
     3001 gatgacgcgg gacaagccgt tttacgtttg gaactgacag aaccgcaacg ttgaaggagc
     3061 cactgagccg cgggtttctg gagtttaatg agctaagcac atacgtcaga aaccattatt
     3121 gcgcgttcaa aagtcgccta aggtcactat cagctagcaa atatttcttg tcaaaaatgc
     3181 tccactgacg ttccataaat tcccctcggt atccaattag agtctcatat tcactctcaa
     3241 ctcgatcgag gcaatggaga acatggagaa cgacgagaac attgtggtgg gacccaagcc
     3301 gttctacccg atcgaggagg gttccgctgg cacccagctg cgcaagtaca tggagcgcta
     3361 cgctaagctg ggtgctatcg ccttcaccaa cgccgtgacc ggtgtggatt actcctacgc
     3421 cgagtacctg gagaagtcct gctgcctcgg aaaggccctg cagaactacg gtctggtcgt
     3481 ggatggccgc attgccctct gctcggagaa ctgcgaggag ttcttcattc ccgtgatcgc
     3541 tggtctgttc atcggagtcg gcgttgcccc aaccaacgag atctacaccc tgcgcgagct
     3601 ggtccacagc ctgggtatca gcaagccgac catcgttttc tcgagcaaga agggcctcga
     3661 caaggttatc accgtgcaaa aaaccgtcac caccatcaag accatcgtta tcctggattc
     3721 caaggttgac taccgcggtt accagtgcct cgacaccttc atcaagcgta acaccccacc
     3781 cggcttccaa gcctcctcct tcaaaaccgt ggaggttgat cgcaaggagc aggtggccct
     3841 gatcatgaac tcctccggct ccaccggtct gccaaagggc gtgcagctga cccacgagaa
     3901 caccgtcacc cgcttctcgc atgcccgtga tcccatctac ggcaaccagg tgtccccagg
     3961 caccgccgtc ctgaccgtcg tgcccctcca ccacggcttc ggcatgttca ccactctcgg
     4021 ctacttgatc tgcggcttcc gtgtggtgat gctgactaag ttcgacgagg agactttcct
     4081 caagaccctg caagactaca agtgcactta cgtgatcctg gtgccgaccc tgttcgccat
     4141 cctgaacaag agcgagctgt tgaacaaata cgacctgtcg aacctggtgg agatcgcttc
     4201 cggcggcgcc ccactgtcga aggaagttgg cgaggccgtt gctcgccgct tcaacctgcc
     4261 cggcgtccgt cagggttacg gcctgaccga gactacctcg gctatcatca tcacccccga
     4321 gggcgacgat aagcccggcg cttccggcaa ggttgttcca ctcttcaaag ccaaggtgat
     4381 cgacctggac accaagaagt ccctgggtcc caaccgtcgc ggagaagtgt gcgtcaaggg
     4441 ccccatgctg atgaagggct acgtcaacaa cccagaggcc accaaggagt tgatcgacga
     4501 ggagggctgg ctgcacactg gtgacatcgg ttactacgac gaagagaagc acttcttcat
     4561 cgttgaccgc ctgaagtcct tgatcaagta caagggctac caggttcccc ccgccgagct
     4621 cgagagcgtc ctgctgcagc acccctccat tttcgatgcc ggagtggccg gtgtccccga
     4681 tcccgtggct ggcgagctgc caggtgccgt cgtcgtgctg gagtccggca agaacatgac
     4741 cgagaaggaa gtgatggatt acgtggcctc gcaagtgtcc aacgccaagc gcctgcgcgg
     4801 cggcgtgcgc ttcgtggatg aggtgcccaa gggcctcacc ggaaagatcg acggtcgcgc
     4861 tatccgcgag atcctcaaga agcccgtggc caagatgtga gcttggaatg gatcttcgat
     4921 cccgatcgtt caaacatttg gcaataaagt ttcttaagat tgaatcctgt tgccggtctt
     4981 gcgacgatta tcatataatt tctgttgaat tacgttaagc atgtaataat taacatgtaa
     5041 tgcatgacgt tatttatgag atgggttttt atgattagag tcccgcaatt atacatttaa
     5101 tacgcgatag aaaacaaaat atagcgcgca aactaggata aattatcgcg cgcggtgtca
     5161 tctatgttac tagatcggga attgccaagc taattcttga agatgaaagg gcctcgtgat
     5221 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     5281 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     5341 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatgggacc gactcgcgct
     5401 gtcatgagac cggatcctga caggatatat tggcgggtaa actaagtcgc tgtatgtgtt
     5461 tgtttgagat ctcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5521 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5581 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5641 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5701 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5761 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5821 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5881 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5941 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     6001 gaagccagtt accttcggaa gaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6061 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6121 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6181 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6241 atgaagtttt aaatcaatct aaagtatata tgtgtaacat tggtctagtg attatttgcc
     6301 gactaccttg gtgatctcgc ctttcacgta gtggacaaat tcttccaact gatctgcgcg
     6361 cgaggccaag cgatcttctt cttgtccaag ataagcctgt ctagcttcaa gtatgacggg
     6421 ctgatactgg gccggcaggc gctccattgc ccagtcggca gcgacatcct tcggcgcgat
     6481 tttgccggtt actgcgctgt accaaatgcg ggacaacgta agcactacat ttcgctcatc
     6541 accagcccag tcgggcggcg agttccatag cgttaaggtt tcatttagcg cctcaaatag
     6601 atcctgttca ggaaccggat caaagagttc ctccgccgct ggacctacca aggcaacgct
     6661 atgttctctt gcttttgtca gcaagatagc cagatcaatg tcgatcgtgg ctggctcgaa
     6721 gatacctgca agaatgtcat tgcgctgcca ttctccaaat tgcagttcgc gcttagctgg
     6781 ataacgccac ggaatgatgt cgtcgtgcac aacaatggtg acttctacag cgcggagaat
     6841 ctcgctctct ccaggggaag ccgaagtttc caaaaggtcg ttgatcaaag ctcgccgcgt
     6901 tgtttcatca agccttacgg tcaccgtaac cagcaaatca atatcactgt gtggcttcag
     6961 gccgccatcc actgcggagc cgtacaaatg tacggccagc aacgtcggtt cgagatggcg
     7021 ctcgatgacg ccaactacct ctgatagttg agtcgatact tcggcgatca ccgcttccct
     7081 cataatgttt aactttgttt tagggcgact gccctgctgc gtaacatcgt tgctgctcca
     7141 taacatcaaa catcgaccca cggcgtaacg cgcttgctgc ttggatgccc gaggcataga
     7201 ctgtacccca aaaaaacagt cataacaagc catgaaaacc gccactgcgc cgttaccacc
     7261 gctgcgttcg gtcaaggttc tggaccagtt gcgtgagcgc atacgctact tgcattacag
     7321 cttacgaacc gaacaggctt atgtccactg ggttcgtgcc ttcatccgtt tccacggtgt
     7381 gcgtcacccg gcaaccttgg gtagcagcga agtcgaggca tttctgtcct ggctggaaca
     7441 ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgatga tatattttta
     7501 tcttgtgcaa tgtaacatca gagattttga gacacaacgt ggctttgttg aataaatcga
     7561 acttttgctg agttgaagga tcagatcacg catcttcccg acaacgcaga ccgttccgtg
     7621 gcaaagcaaa agttcaaaat caccaactgg tccacctaca acaaagctct catcaaccgt
     7681 ggctccctca ctttctggct ggatgatggg gcgattcagg cgatccccat ccaacagccc
     7741 gccgtcgagc gggctttttt atccccggaa gcctgtggat agagggtagt tatccacgtg
     7801 aaaccgctaa tgccccgcaa agccttgatt cacggggctt tccggcccgc tccaaaaact
     7861 atccacgtga aatcgctaat cagggtacgt gaaatcgcta atcggagtac gtgaaatcgc
     7921 taataaggtc acgtgaaatc gctaatcaaa aaggcacgtg agaacgctaa tagccctttc
     7981 agatcaacag cttgcaaaca cccctcgctc cggcaagtag ttacagcaag tagtatgttc
     8041 aattagcttt tcaattatga atatatatat caattattgg tcgcccttgg cttgtggaca
     8101 atgcgctacg cgcaccggct ccgcccgtgg acaaccgcaa gcggttgccc accgtcgagc
     8161 gcctttgccc acaacccggc ggccggccgc aacagatcgt tttataaatt tttttttttg
     8221 aaaaagaaaa agcccgaaag gcggcaacct ctcgggcttc tggatttccg atccccggaa
     8281 ttagatcttg gcaggatata ttgtggtg
//
