LOCUS       sequence-334465-        8137 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid Double Transcriptional Unit - 
            pCmYLCV911_E-Luc_AtACT2t+pNOS_Red-F_NOSt.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8137)
  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 8137)
  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: pGREAT15
FEATURES             Location/Qualifiers
     source          1..8137
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             1960..1968
                     /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        2869..3048
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      4753..5005
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(5040..5058)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5247..5271
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5362..5950)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5442..5461)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6121..6912)
                     /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      7556..7991
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    join(8118..8137,1..3)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
ORIGIN
        1 taacggatcc ggtctcagga gtggcagaca tactgtccca caaatgaaga tggaatctgt
       61 aaaagaaaac gcgtgaaata atgcgtctga caaaggttag gtcggctgcc tttaatcaat
      121 accaaagtgg tccctaccac gatggaaaaa ctgtgcagtc ggtttggctt tttctgacga
      181 acaaataaga ttcgtggccg acaggtgggg gtccaccatg tgaaggcatc ttcagactcc
      241 aataatggag caatgacgta agggcttacg aaataagtaa gggtagtttg ggaaatgtcc
      301 actcacccgt cagtctataa atacttagcc cctccctcat tgttaaggga gcaaaatctc
      361 agagagatag tcctagagag agaaagagag caagtagcct agaagtagtc aaggcggcga
      421 agtattcagg caggtggcca ggaagaagaa aagccaagac gacgaaaaca ggtaagagct
      481 aagcttaatg gagagagaga agaacgtggt gtacggcccc gagcccaagc accctctggg
      541 caacttcacc gccggcgaga tgctgtacaa cgctctgcac aagcactccc acatccccca
      601 ggccatcctg gacgtgatgg gcaacgagtc cctttcctac caggagttct tcgacactac
      661 tgtgaagctg ggccagagcc tccagaactg tggctacaag atgaacgatg tcgtgtcgat
      721 ctgtgcagag aacaacaaga gattcttcat ccccatcatc tccgcctggt acatcggcat
      781 ggtggtggcc cctgtgaacg aggactatat cccagacgag ctgtgtaaag tgaccggcat
      841 ctccaagccg atcctggtct ttaccactag gaagatcctg cctaaggttt tggaggttaa
      901 agacagaacc aactacataa agaggatcat catactggac tctgaagaga acctgctggg
      961 ctgcgagagc ctgcacaact tcatgtccag gtactccgac aacaacctcc aaacattcaa
     1021 gcctctgcac tacgaccctg tggaccaggt agccgccatc ctgtgctcct ccggcacaac
     1081 cggcctgcct aaaggcgtga tgcagaccca caggaacatc tgtgtgagac tcacacacgc
     1141 atctgacccc agagtgggta cacaactcat ccccggcgta tccgtgctgg cctacctgcc
     1201 attcttccac gccttcggct tcagtatcaa cctgggctat ttcatggtgg gcctgagagt
     1261 ggtgatgctc cgaaggttta accaggaggt gttcctgaag gccatccagg actacgaggt
     1321 gaggagcgtg atcaacgttc cctccacaat cctgttcctg tccaagagcc ctctggtgga
     1381 caagtacgac ctatccaccc tggcggagct gtgctgtgga gccgctcctc tggcgaagga
     1441 ggtggccgag atcgccgtga agaggctgaa cctgccaggg atacggtgtg gctacggtct
     1501 aacagagtct acctccgcca acatccatac tctgcacaac gagttcaagt ccggctccct
     1561 gggcaaggtg acaccttaca tggccgccaa gatcatcgac aggaacaccg gcgaggccct
     1621 gggtccaaac caggtgggcg agctgtgcat ctggggacct atggtaacaa aaggctatgt
     1681 gaacaaccca caggctacta aggaggccat cgacgacgac ggctggctgc actctggcga
     1741 cttcggctac tacgacgagg acgagtattt ctacatcgtg gaccggtaca aggagctgat
     1801 caaatacaag ggctatcagg tcgcccctgt ggagctggag gagatcctcc ttcagcaccc
     1861 aggcatcagg gacgtggccg tcgtgggtat ccctgacatc gaggccggcg agctgccagc
     1921 cggcttcgtg gtgaagcagc ccggcgccca actcaccgct aaggaggtgt acgacttcct
     1981 ggcccagagg gtgtctcact ccaagtacct gaggggcggc gtaaggttcg tggactctat
     2041 ccccaggaac gtgacaggca agattagtcg aaaagagctg agggaggccc tgatggagaa
     2101 ggcttctaag ctgtaagctt gctctcaaga tcaaaggctt aaaaagctgg ggttttatga
     2161 atgggatcaa agtttctttt tttcttttat atttgcttct ccatttgttt gtttcatttc
     2221 cctttttgtt ttcgtttcta tgatgcactt gtgtgtgaca aactctctgg gtttttactt
     2281 acgtctgcgt ttcaaaaaaa aaaaccgctt tcgttttgcg ttttagtccc attgttttgt
     2341 agctctgagt gatcgaattg atgcctcttt attccttttg ttccctataa tttctttcaa
     2401 aactcagaag aaaaaccttg aaactctttg caatgttaat ataagtattg tataagattt
     2461 ttattgattt ggttattagt cttacttttg ctacctccat cttcacttgg aactgatatt
     2521 ctgaatagtt aaagcgttac atgtcctcca ttcacaaatg aacttaaact agcacaaagt
     2581 cagatatttt aagatcgcac cattttatat aaccccaatc gtcaattcta ctgtttcaag
     2641 ttttacacca aaacaattac gaggtgtatc tattcgttac tcttttcgct tattaatcct
     2701 tttacatgat gcaaaatata acaaattaat ttcaaattgt cctgcacatt tggtttatac
     2761 atcttgattc caaaaagtcg ctgtcaggag atacatgaga attaagggag tcacggtatg
     2821 acccccgccg atgacgcggg acaagccgtt ttacgtttgg aactgacaga accgcaacgt
     2881 tgaaggagcc actgagccgc gggtttctgg agtttaatga gctaagcaca tacgtcagaa
     2941 accattattg cgcgttcaaa agtcgcctaa ggtcactatc agctagcaaa tatttcttgt
     3001 caaaaatgct ccactgacgt tccataaatt cccctcggta tccaattaga gtctcatatt
     3061 cactctcaac tcgatcgagg caatggagaa catggagaac gacgagaaca ttgtggtggg
     3121 acccaagccg ttctacccga tcgaggaggg ttccgctggc acccagctgc gcaagtacat
     3181 ggagcgctac gctaagctgg gtgctatcgc cttcaccaac gccgtgaccg gtgtggatta
     3241 ctcctacgcc gagtacctgg agaagtcctg ctgcctcgga aaggccctgc agaactacgg
     3301 tctggtcgtg gatggccgca ttgccctctg ctcggagaac tgcgaggagt tcttcattcc
     3361 cgtgatcgct ggtctgttca tcggagtcgg cgttgcccca accaacgaga tctacaccct
     3421 gcgcgagctg gtccacagcc tgggtatcag caagccgacc atcgttttct cgagcaagaa
     3481 gggcctcgac aaggttatca ccgtgcaaaa aaccgtcacc accatcaaga ccatcgttat
     3541 cctggattcc aaggttgact accgcggtta ccagtgcctc gacaccttca tcaagcgtaa
     3601 caccccaccc ggcttccaag cctcctcctt caaaaccgtg gaggttgatc gcaaggagca
     3661 ggtggccctg atcatgaact cctccggctc caccggtctg ccaaagggcg tgcagctgac
     3721 ccacgagaac accgtcaccc gcttctcgca tgcccgtgat cccatctacg gcaaccaggt
     3781 gtccccaggc accgccgtcc tgaccgtcgt gcccctccac cacggcttcg gcatgttcac
     3841 cactctcggc tacttgatct gcggcttccg tgtggtgatg ctgactaagt tcgacgagga
     3901 gactttcctc aagaccctgc aagactacaa gtgcacttac gtgatcctgg tgccgaccct
     3961 gttcgccatc ctgaacaaga gcgagctgtt gaacaaatac gacctgtcga acctggtgga
     4021 gatcgcttcc ggcggcgccc cactgtcgaa ggaagttggc gaggccgttg ctcgccgctt
     4081 caacctgccc ggcgtccgtc agggttacgg cctgaccgag actacctcgg ctatcatcat
     4141 cacccccgag ggcgacgata agcccggcgc ttccggcaag gttgttccac tcttcaaagc
     4201 caaggtgatc gacctggaca ccaagaagtc cctgggtccc aaccgtcgcg gagaagtgtg
     4261 cgtcaagggc cccatgctga tgaagggcta cgtcaacaac ccagaggcca ccaaggagtt
     4321 gatcgacgag gagggctggc tgcacactgg tgacatcggt tactacgacg aagagaagca
     4381 cttcttcatc gttgaccgcc tgaagtcctt gatcaagtac aagggctacc aggttccccc
     4441 cgccgagctc gagagcgtcc tgctgcagca cccctccatt ttcgatgccg gagtggccgg
     4501 tgtccccgat cccgtggctg gcgagctgcc aggtgccgtc gtcgtgctgg agtccggcaa
     4561 gaacatgacc gagaaggaag tgatggatta cgtggcctcg caagtgtcca acgccaagcg
     4621 cctgcgcggc ggcgtgcgct tcgtggatga ggtgcccaag ggcctcaccg gaaagatcga
     4681 cggtcgcgct atccgcgaga tcctcaagaa gcccgtggcc aagatgtgag cttggaatgg
     4741 atcttcgatc ccgatcgttc aaacatttgg caataaagtt tcttaagatt gaatcctgtt
     4801 gccggtcttg cgacgattat catataattt ctgttgaatt acgttaagca tgtaataatt
     4861 aacatgtaat gcatgacgtt atttatgaga tgggttttta tgattagagt cccgcaatta
     4921 tacatttaat acgcgataga aaacaaaata tagcgcgcaa actaggataa attatcgcgc
     4981 gcggtgtcat ctatgttact agatcgggaa ttgccaagct aattcttgaa gatgaaaggg
     5041 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     5101 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     5161 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatgggaccg
     5221 actcgcgctg tcatgagacc ggatcctgac aggatatatt ggcgggtaaa ctaagtcgct
     5281 gtatgtgttt gtttgagatc tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5341 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5401 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5461 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5521 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5581 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5641 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5701 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5761 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5821 cgctctgctg aagccagtta ccttcggaag aagagttggt agctcttgat ccggcaaaca
     5881 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5941 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6001 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6061 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gtgtaacatt ggtctagtga
     6121 ttatttgccg actaccttgg tgatctcgcc tttcacgtag tggacaaatt cttccaactg
     6181 atctgcgcgc gaggccaagc gatcttcttc ttgtccaaga taagcctgtc tagcttcaag
     6241 tatgacgggc tgatactggg ccggcaggcg ctccattgcc cagtcggcag cgacatcctt
     6301 cggcgcgatt ttgccggtta ctgcgctgta ccaaatgcgg gacaacgtaa gcactacatt
     6361 tcgctcatca ccagcccagt cgggcggcga gttccatagc gttaaggttt catttagcgc
     6421 ctcaaataga tcctgttcag gaaccggatc aaagagttcc tccgccgctg gacctaccaa
     6481 ggcaacgcta tgttctcttg cttttgtcag caagatagcc agatcaatgt cgatcgtggc
     6541 tggctcgaag atacctgcaa gaatgtcatt gcgctgccat tctccaaatt gcagttcgcg
     6601 cttagctgga taacgccacg gaatgatgtc gtcgtgcaca acaatggtga cttctacagc
     6661 gcggagaatc tcgctctctc caggggaagc cgaagtttcc aaaaggtcgt tgatcaaagc
     6721 tcgccgcgtt gtttcatcaa gccttacggt caccgtaacc agcaaatcaa tatcactgtg
     6781 tggcttcagg ccgccatcca ctgcggagcc gtacaaatgt acggccagca acgtcggttc
     6841 gagatggcgc tcgatgacgc caactacctc tgatagttga gtcgatactt cggcgatcac
     6901 cgcttccctc ataatgttta actttgtttt agggcgactg ccctgctgcg taacatcgtt
     6961 gctgctccat aacatcaaac atcgacccac ggcgtaacgc gcttgctgct tggatgcccg
     7021 aggcatagac tgtaccccaa aaaaacagtc ataacaagcc atgaaaaccg ccactgcgcc
     7081 gttaccaccg ctgcgttcgg tcaaggttct ggaccagttg cgtgagcgca tacgctactt
     7141 gcattacagc ttacgaaccg aacaggctta tgtccactgg gttcgtgcct tcatccgttt
     7201 ccacggtgtg cgtcacccgg caaccttggg tagcagcgaa gtcgaggcat ttctgtcctg
     7261 gctggaacac cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat
     7321 atatttttat cttgtgcaat gtaacatcag agattttgag acacaacgtg gctttgttga
     7381 ataaatcgaa cttttgctga gttgaaggat cagatcacgc atcttcccga caacgcagac
     7441 cgttccgtgg caaagcaaaa gttcaaaatc accaactggt ccacctacaa caaagctctc
     7501 atcaaccgtg gctccctcac tttctggctg gatgatgggg cgattcaggc gatccccatc
     7561 caacagcccg ccgtcgagcg ggctttttta tccccggaag cctgtggata gagggtagtt
     7621 atccacgtga aaccgctaat gccccgcaaa gccttgattc acggggcttt ccggcccgct
     7681 ccaaaaacta tccacgtgaa atcgctaatc agggtacgtg aaatcgctaa tcggagtacg
     7741 tgaaatcgct aataaggtca cgtgaaatcg ctaatcaaaa aggcacgtga gaacgctaat
     7801 agccctttca gatcaacagc ttgcaaacac ccctcgctcc ggcaagtagt tacagcaagt
     7861 agtatgttca attagctttt caattatgaa tatatatatc aattattggt cgcccttggc
     7921 ttgtggacaa tgcgctacgc gcaccggctc cgcccgtgga caaccgcaag cggttgccca
     7981 ccgtcgagcg cctttgccca caacccggcg gccggccgca acagatcgtt ttataaattt
     8041 ttttttttga aaaagaaaaa gcccgaaagg cggcaacctc tcgggcttct ggatttccga
     8101 tccccggaat tagatcttgg caggatatat tgtggtg
//
