LOCUS       sequence-334552-        7915 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid Double Transcriptional Unit - 
            pG10-90_E-Luc_AtACT2t+pNOS_Red-F_NOSt.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7915)
  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 7915)
  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: pGREAT14
FEATURES             Location/Qualifiers
     source          1..7915
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     196..216
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     promoter        205..251
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     RBS             1738..1746
                     /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        2647..2826
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      4531..4783
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4818..4836)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5025..5049
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5140..5728)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5220..5239)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5899..6690)
                     /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      7334..7769
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    join(7896..7915,1..3)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
ORIGIN
        1 taacggatcc ggtctcagga gatagtttaa actgaaggcg ggaaacgaca atctgatcca
       61 agctcaagct aagcttgcat gcctgcagga tatcgtggat ccaagcttgc cacgtgccgc
      121 cacgtgccgc cacgtgccgc cacgtgcctc tagaggatcc atctccactg acgtaaggga
      181 tgacgcacaa tcccactatc cttcgcaaga cccttcctct atataaggaa gttcatttca
      241 tttggagagg acacgctggg atccaatgga gagagagaag aacgtggtgt acggccccga
      301 gcccaagcac cctctgggca acttcaccgc cggcgagatg ctgtacaacg ctctgcacaa
      361 gcactcccac atcccccagg ccatcctgga cgtgatgggc aacgagtccc tttcctacca
      421 ggagttcttc gacactactg tgaagctggg ccagagcctc cagaactgtg gctacaagat
      481 gaacgatgtc gtgtcgatct gtgcagagaa caacaagaga ttcttcatcc ccatcatctc
      541 cgcctggtac atcggcatgg tggtggcccc tgtgaacgag gactatatcc cagacgagct
      601 gtgtaaagtg accggcatct ccaagccgat cctggtcttt accactagga agatcctgcc
      661 taaggttttg gaggttaaag acagaaccaa ctacataaag aggatcatca tactggactc
      721 tgaagagaac ctgctgggct gcgagagcct gcacaacttc atgtccaggt actccgacaa
      781 caacctccaa acattcaagc ctctgcacta cgaccctgtg gaccaggtag ccgccatcct
      841 gtgctcctcc ggcacaaccg gcctgcctaa aggcgtgatg cagacccaca ggaacatctg
      901 tgtgagactc acacacgcat ctgaccccag agtgggtaca caactcatcc ccggcgtatc
      961 cgtgctggcc tacctgccat tcttccacgc cttcggcttc agtatcaacc tgggctattt
     1021 catggtgggc ctgagagtgg tgatgctccg aaggtttaac caggaggtgt tcctgaaggc
     1081 catccaggac tacgaggtga ggagcgtgat caacgttccc tccacaatcc tgttcctgtc
     1141 caagagccct ctggtggaca agtacgacct atccaccctg gcggagctgt gctgtggagc
     1201 cgctcctctg gcgaaggagg tggccgagat cgccgtgaag aggctgaacc tgccagggat
     1261 acggtgtggc tacggtctaa cagagtctac ctccgccaac atccatactc tgcacaacga
     1321 gttcaagtcc ggctccctgg gcaaggtgac accttacatg gccgccaaga tcatcgacag
     1381 gaacaccggc gaggccctgg gtccaaacca ggtgggcgag ctgtgcatct ggggacctat
     1441 ggtaacaaaa ggctatgtga acaacccaca ggctactaag gaggccatcg acgacgacgg
     1501 ctggctgcac tctggcgact tcggctacta cgacgaggac gagtatttct acatcgtgga
     1561 ccggtacaag gagctgatca aatacaaggg ctatcaggtc gcccctgtgg agctggagga
     1621 gatcctcctt cagcacccag gcatcaggga cgtggccgtc gtgggtatcc ctgacatcga
     1681 ggccggcgag ctgccagccg gcttcgtggt gaagcagccc ggcgcccaac tcaccgctaa
     1741 ggaggtgtac gacttcctgg cccagagggt gtctcactcc aagtacctga ggggcggcgt
     1801 aaggttcgtg gactctatcc ccaggaacgt gacaggcaag attagtcgaa aagagctgag
     1861 ggaggccctg atggagaagg cttctaagct gtaagcttgc tctcaagatc aaaggcttaa
     1921 aaagctgggg ttttatgaat gggatcaaag tttctttttt tcttttatat ttgcttctcc
     1981 atttgtttgt ttcatttccc tttttgtttt cgtttctatg atgcacttgt gtgtgacaaa
     2041 ctctctgggt ttttacttac gtctgcgttt caaaaaaaaa aaccgctttc gttttgcgtt
     2101 ttagtcccat tgttttgtag ctctgagtga tcgaattgat gcctctttat tccttttgtt
     2161 ccctataatt tctttcaaaa ctcagaagaa aaaccttgaa actctttgca atgttaatat
     2221 aagtattgta taagattttt attgatttgg ttattagtct tacttttgct acctccatct
     2281 tcacttggaa ctgatattct gaatagttaa agcgttacat gtcctccatt cacaaatgaa
     2341 cttaaactag cacaaagtca gatattttaa gatcgcacca ttttatataa ccccaatcgt
     2401 caattctact gtttcaagtt ttacaccaaa acaattacga ggtgtatcta ttcgttactc
     2461 ttttcgctta ttaatccttt tacatgatgc aaaatataac aaattaattt caaattgtcc
     2521 tgcacatttg gtttatacat cttgattcca aaaagtcgct gtcaggagat acatgagaat
     2581 taagggagtc acggtatgac ccccgccgat gacgcgggac aagccgtttt acgtttggaa
     2641 ctgacagaac cgcaacgttg aaggagccac tgagccgcgg gtttctggag tttaatgagc
     2701 taagcacata cgtcagaaac cattattgcg cgttcaaaag tcgcctaagg tcactatcag
     2761 ctagcaaata tttcttgtca aaaatgctcc actgacgttc cataaattcc cctcggtatc
     2821 caattagagt ctcatattca ctctcaactc gatcgaggca atggagaaca tggagaacga
     2881 cgagaacatt gtggtgggac ccaagccgtt ctacccgatc gaggagggtt ccgctggcac
     2941 ccagctgcgc aagtacatgg agcgctacgc taagctgggt gctatcgcct tcaccaacgc
     3001 cgtgaccggt gtggattact cctacgccga gtacctggag aagtcctgct gcctcggaaa
     3061 ggccctgcag aactacggtc tggtcgtgga tggccgcatt gccctctgct cggagaactg
     3121 cgaggagttc ttcattcccg tgatcgctgg tctgttcatc ggagtcggcg ttgccccaac
     3181 caacgagatc tacaccctgc gcgagctggt ccacagcctg ggtatcagca agccgaccat
     3241 cgttttctcg agcaagaagg gcctcgacaa ggttatcacc gtgcaaaaaa ccgtcaccac
     3301 catcaagacc atcgttatcc tggattccaa ggttgactac cgcggttacc agtgcctcga
     3361 caccttcatc aagcgtaaca ccccacccgg cttccaagcc tcctccttca aaaccgtgga
     3421 ggttgatcgc aaggagcagg tggccctgat catgaactcc tccggctcca ccggtctgcc
     3481 aaagggcgtg cagctgaccc acgagaacac cgtcacccgc ttctcgcatg cccgtgatcc
     3541 catctacggc aaccaggtgt ccccaggcac cgccgtcctg accgtcgtgc ccctccacca
     3601 cggcttcggc atgttcacca ctctcggcta cttgatctgc ggcttccgtg tggtgatgct
     3661 gactaagttc gacgaggaga ctttcctcaa gaccctgcaa gactacaagt gcacttacgt
     3721 gatcctggtg ccgaccctgt tcgccatcct gaacaagagc gagctgttga acaaatacga
     3781 cctgtcgaac ctggtggaga tcgcttccgg cggcgcccca ctgtcgaagg aagttggcga
     3841 ggccgttgct cgccgcttca acctgcccgg cgtccgtcag ggttacggcc tgaccgagac
     3901 tacctcggct atcatcatca cccccgaggg cgacgataag cccggcgctt ccggcaaggt
     3961 tgttccactc ttcaaagcca aggtgatcga cctggacacc aagaagtccc tgggtcccaa
     4021 ccgtcgcgga gaagtgtgcg tcaagggccc catgctgatg aagggctacg tcaacaaccc
     4081 agaggccacc aaggagttga tcgacgagga gggctggctg cacactggtg acatcggtta
     4141 ctacgacgaa gagaagcact tcttcatcgt tgaccgcctg aagtccttga tcaagtacaa
     4201 gggctaccag gttccccccg ccgagctcga gagcgtcctg ctgcagcacc cctccatttt
     4261 cgatgccgga gtggccggtg tccccgatcc cgtggctggc gagctgccag gtgccgtcgt
     4321 cgtgctggag tccggcaaga acatgaccga gaaggaagtg atggattacg tggcctcgca
     4381 agtgtccaac gccaagcgcc tgcgcggcgg cgtgcgcttc gtggatgagg tgcccaaggg
     4441 cctcaccgga aagatcgacg gtcgcgctat ccgcgagatc ctcaagaagc ccgtggccaa
     4501 gatgtgagct tggaatggat cttcgatccc gatcgttcaa acatttggca ataaagtttc
     4561 ttaagattga atcctgttgc cggtcttgcg acgattatca tataatttct gttgaattac
     4621 gttaagcatg taataattaa catgtaatgc atgacgttat ttatgagatg ggtttttatg
     4681 attagagtcc cgcaattata catttaatac gcgatagaaa acaaaatata gcgcgcaaac
     4741 taggataaat tatcgcgcgc ggtgtcatct atgttactag atcgggaatt gccaagctaa
     4801 ttcttgaaga tgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     4861 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4921 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4981 gcttcaataa tgggaccgac tcgcgctgtc atgagaccgg atcctgacag gatatattgg
     5041 cgggtaaact aagtcgctgt atgtgtttgt ttgagatctc atgtgagcaa aaggccagca
     5101 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5161 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5221 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5281 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5341 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5401 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5461 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5521 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5581 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaagaa gagttggtag
     5641 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5701 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5761 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5821 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatgt
     5881 gtaacattgg tctagtgatt atttgccgac taccttggtg atctcgcctt tcacgtagtg
     5941 gacaaattct tccaactgat ctgcgcgcga ggccaagcga tcttcttctt gtccaagata
     6001 agcctgtcta gcttcaagta tgacgggctg atactgggcc ggcaggcgct ccattgccca
     6061 gtcggcagcg acatccttcg gcgcgatttt gccggttact gcgctgtacc aaatgcggga
     6121 caacgtaagc actacatttc gctcatcacc agcccagtcg ggcggcgagt tccatagcgt
     6181 taaggtttca tttagcgcct caaatagatc ctgttcagga accggatcaa agagttcctc
     6241 cgccgctgga cctaccaagg caacgctatg ttctcttgct tttgtcagca agatagccag
     6301 atcaatgtcg atcgtggctg gctcgaagat acctgcaaga atgtcattgc gctgccattc
     6361 tccaaattgc agttcgcgct tagctggata acgccacgga atgatgtcgt cgtgcacaac
     6421 aatggtgact tctacagcgc ggagaatctc gctctctcca ggggaagccg aagtttccaa
     6481 aaggtcgttg atcaaagctc gccgcgttgt ttcatcaagc cttacggtca ccgtaaccag
     6541 caaatcaata tcactgtgtg gcttcaggcc gccatccact gcggagccgt acaaatgtac
     6601 ggccagcaac gtcggttcga gatggcgctc gatgacgcca actacctctg atagttgagt
     6661 cgatacttcg gcgatcaccg cttccctcat aatgtttaac tttgttttag ggcgactgcc
     6721 ctgctgcgta acatcgttgc tgctccataa catcaaacat cgacccacgg cgtaacgcgc
     6781 ttgctgcttg gatgcccgag gcatagactg taccccaaaa aaacagtcat aacaagccat
     6841 gaaaaccgcc actgcgccgt taccaccgct gcgttcggtc aaggttctgg accagttgcg
     6901 tgagcgcata cgctacttgc attacagctt acgaaccgaa caggcttatg tccactgggt
     6961 tcgtgccttc atccgtttcc acggtgtgcg tcacccggca accttgggta gcagcgaagt
     7021 cgaggcattt ctgtcctggc tggaacaccc cttgtattac tgtttatgta agcagacagt
     7081 tttattgttc atgatgatat atttttatct tgtgcaatgt aacatcagag attttgagac
     7141 acaacgtggc tttgttgaat aaatcgaact tttgctgagt tgaaggatca gatcacgcat
     7201 cttcccgaca acgcagaccg ttccgtggca aagcaaaagt tcaaaatcac caactggtcc
     7261 acctacaaca aagctctcat caaccgtggc tccctcactt tctggctgga tgatggggcg
     7321 attcaggcga tccccatcca acagcccgcc gtcgagcggg cttttttatc cccggaagcc
     7381 tgtggataga gggtagttat ccacgtgaaa ccgctaatgc cccgcaaagc cttgattcac
     7441 ggggctttcc ggcccgctcc aaaaactatc cacgtgaaat cgctaatcag ggtacgtgaa
     7501 atcgctaatc ggagtacgtg aaatcgctaa taaggtcacg tgaaatcgct aatcaaaaag
     7561 gcacgtgaga acgctaatag ccctttcaga tcaacagctt gcaaacaccc ctcgctccgg
     7621 caagtagtta cagcaagtag tatgttcaat tagcttttca attatgaata tatatatcaa
     7681 ttattggtcg cccttggctt gtggacaatg cgctacgcgc accggctccg cccgtggaca
     7741 accgcaagcg gttgcccacc gtcgagcgcc tttgcccaca acccggcggc cggccgcaac
     7801 agatcgtttt ataaattttt ttttttgaaa aagaaaaagc ccgaaaggcg gcaacctctc
     7861 gggcttctgg atttccgatc cccggaatta gatcttggca ggatatattg tggtg
//
