LOCUS       sequence-335140-        7876 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid Double Transcriptional Unit - 
            pAtUBQ10_E-Luc_35St+pNOS_Red-F_NOSt.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7876)
  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 7876)
  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: pGREAT23
FEATURES             Location/Qualifiers
     source          1..7876
                     /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        2608..2787
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      4492..4744
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4779..4797)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4986..5010
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5101..5689)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5181..5200)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5860..6651)
                     /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      7295..7730
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    join(7857..7876,1..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 tcggccatgc tagagtccgc aaaaatcacc agtctctctc tacaaatcta
     2341 tctctctcta tttttctcca gaataatgtg tgagtagttc ccagataagg gaattagggt
     2401 tcttataggg tttcgctcat gtgttgagca tataagaaac ccttagtatg tatttgtatt
     2461 tgtaaaatac ttctatcaat aaaatttcta attcctaaaa ccaaaatcca gtgacctcgc
     2521 tgtcaggaga tacatgagaa ttaagggagt cacggtatga cccccgccga tgacgcggga
     2581 caagccgttt tacgtttgga actgacagaa ccgcaacgtt gaaggagcca ctgagccgcg
     2641 ggtttctgga gtttaatgag ctaagcacat acgtcagaaa ccattattgc gcgttcaaaa
     2701 gtcgcctaag gtcactatca gctagcaaat atttcttgtc aaaaatgctc cactgacgtt
     2761 ccataaattc ccctcggtat ccaattagag tctcatattc actctcaact cgatcgaggc
     2821 aatggagaac atggagaacg acgagaacat tgtggtggga cccaagccgt tctacccgat
     2881 cgaggagggt tccgctggca cccagctgcg caagtacatg gagcgctacg ctaagctggg
     2941 tgctatcgcc ttcaccaacg ccgtgaccgg tgtggattac tcctacgccg agtacctgga
     3001 gaagtcctgc tgcctcggaa aggccctgca gaactacggt ctggtcgtgg atggccgcat
     3061 tgccctctgc tcggagaact gcgaggagtt cttcattccc gtgatcgctg gtctgttcat
     3121 cggagtcggc gttgccccaa ccaacgagat ctacaccctg cgcgagctgg tccacagcct
     3181 gggtatcagc aagccgacca tcgttttctc gagcaagaag ggcctcgaca aggttatcac
     3241 cgtgcaaaaa accgtcacca ccatcaagac catcgttatc ctggattcca aggttgacta
     3301 ccgcggttac cagtgcctcg acaccttcat caagcgtaac accccacccg gcttccaagc
     3361 ctcctccttc aaaaccgtgg aggttgatcg caaggagcag gtggccctga tcatgaactc
     3421 ctccggctcc accggtctgc caaagggcgt gcagctgacc cacgagaaca ccgtcacccg
     3481 cttctcgcat gcccgtgatc ccatctacgg caaccaggtg tccccaggca ccgccgtcct
     3541 gaccgtcgtg cccctccacc acggcttcgg catgttcacc actctcggct acttgatctg
     3601 cggcttccgt gtggtgatgc tgactaagtt cgacgaggag actttcctca agaccctgca
     3661 agactacaag tgcacttacg tgatcctggt gccgaccctg ttcgccatcc tgaacaagag
     3721 cgagctgttg aacaaatacg acctgtcgaa cctggtggag atcgcttccg gcggcgcccc
     3781 actgtcgaag gaagttggcg aggccgttgc tcgccgcttc aacctgcccg gcgtccgtca
     3841 gggttacggc ctgaccgaga ctacctcggc tatcatcatc acccccgagg gcgacgataa
     3901 gcccggcgct tccggcaagg ttgttccact cttcaaagcc aaggtgatcg acctggacac
     3961 caagaagtcc ctgggtccca accgtcgcgg agaagtgtgc gtcaagggcc ccatgctgat
     4021 gaagggctac gtcaacaacc cagaggccac caaggagttg atcgacgagg agggctggct
     4081 gcacactggt gacatcggtt actacgacga agagaagcac ttcttcatcg ttgaccgcct
     4141 gaagtccttg atcaagtaca agggctacca ggttcccccc gccgagctcg agagcgtcct
     4201 gctgcagcac ccctccattt tcgatgccgg agtggccggt gtccccgatc ccgtggctgg
     4261 cgagctgcca ggtgccgtcg tcgtgctgga gtccggcaag aacatgaccg agaaggaagt
     4321 gatggattac gtggcctcgc aagtgtccaa cgccaagcgc ctgcgcggcg gcgtgcgctt
     4381 cgtggatgag gtgcccaagg gcctcaccgg aaagatcgac ggtcgcgcta tccgcgagat
     4441 cctcaagaag cccgtggcca agatgtgagc ttggaatgga tcttcgatcc cgatcgttca
     4501 aacatttggc aataaagttt cttaagattg aatcctgttg ccggtcttgc gacgattatc
     4561 atataatttc tgttgaatta cgttaagcat gtaataatta acatgtaatg catgacgtta
     4621 tttatgagat gggtttttat gattagagtc ccgcaattat acatttaata cgcgatagaa
     4681 aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg cggtgtcatc tatgttacta
     4741 gatcgggaat tgccaagcta attcttgaag atgaaagggc ctcgtgatac gcctattttt
     4801 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     4861 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4921 gagacaataa ccctgataaa tgcttcaata atgggaccga ctcgcgctgt catgagaccg
     4981 gatcctgaca ggatatattg gcgggtaaac taagtcgctg tatgtgtttg tttgagatct
     5041 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5101 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5161 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5221 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5281 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5341 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5401 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5461 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5521 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5581 cttcggaaga agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5641 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5701 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5761 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5821 atcaatctaa agtatatatg tgtaacattg gtctagtgat tatttgccga ctaccttggt
     5881 gatctcgcct ttcacgtagt ggacaaattc ttccaactga tctgcgcgcg aggccaagcg
     5941 atcttcttct tgtccaagat aagcctgtct agcttcaagt atgacgggct gatactgggc
     6001 cggcaggcgc tccattgccc agtcggcagc gacatccttc ggcgcgattt tgccggttac
     6061 tgcgctgtac caaatgcggg acaacgtaag cactacattt cgctcatcac cagcccagtc
     6121 gggcggcgag ttccatagcg ttaaggtttc atttagcgcc tcaaatagat cctgttcagg
     6181 aaccggatca aagagttcct ccgccgctgg acctaccaag gcaacgctat gttctcttgc
     6241 ttttgtcagc aagatagcca gatcaatgtc gatcgtggct ggctcgaaga tacctgcaag
     6301 aatgtcattg cgctgccatt ctccaaattg cagttcgcgc ttagctggat aacgccacgg
     6361 aatgatgtcg tcgtgcacaa caatggtgac ttctacagcg cggagaatct cgctctctcc
     6421 aggggaagcc gaagtttcca aaaggtcgtt gatcaaagct cgccgcgttg tttcatcaag
     6481 ccttacggtc accgtaacca gcaaatcaat atcactgtgt ggcttcaggc cgccatccac
     6541 tgcggagccg tacaaatgta cggccagcaa cgtcggttcg agatggcgct cgatgacgcc
     6601 aactacctct gatagttgag tcgatacttc ggcgatcacc gcttccctca taatgtttaa
     6661 ctttgtttta gggcgactgc cctgctgcgt aacatcgttg ctgctccata acatcaaaca
     6721 tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact gtaccccaaa
     6781 aaaacagtca taacaagcca tgaaaaccgc cactgcgccg ttaccaccgc tgcgttcggt
     6841 caaggttctg gaccagttgc gtgagcgcat acgctacttg cattacagct tacgaaccga
     6901 acaggcttat gtccactggg ttcgtgcctt catccgtttc cacggtgtgc gtcacccggc
     6961 aaccttgggt agcagcgaag tcgaggcatt tctgtcctgg ctggaacacc ccttgtatta
     7021 ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc ttgtgcaatg
     7081 taacatcaga gattttgaga cacaacgtgg ctttgttgaa taaatcgaac ttttgctgag
     7141 ttgaaggatc agatcacgca tcttcccgac aacgcagacc gttccgtggc aaagcaaaag
     7201 ttcaaaatca ccaactggtc cacctacaac aaagctctca tcaaccgtgg ctccctcact
     7261 ttctggctgg atgatggggc gattcaggcg atccccatcc aacagcccgc cgtcgagcgg
     7321 gcttttttat ccccggaagc ctgtggatag agggtagtta tccacgtgaa accgctaatg
     7381 ccccgcaaag ccttgattca cggggctttc cggcccgctc caaaaactat ccacgtgaaa
     7441 tcgctaatca gggtacgtga aatcgctaat cggagtacgt gaaatcgcta ataaggtcac
     7501 gtgaaatcgc taatcaaaaa ggcacgtgag aacgctaata gccctttcag atcaacagct
     7561 tgcaaacacc cctcgctccg gcaagtagtt acagcaagta gtatgttcaa ttagcttttc
     7621 aattatgaat atatatatca attattggtc gcccttggct tgtggacaat gcgctacgcg
     7681 caccggctcc gcccgtggac aaccgcaagc ggttgcccac cgtcgagcgc ctttgcccac
     7741 aacccggcgg ccggccgcaa cagatcgttt tataaatttt tttttttgaa aaagaaaaag
     7801 cccgaaaggc ggcaacctct cgggcttctg gatttccgat ccccggaatt agatcttggc
     7861 aggatatatt gtggtg
//
