LOCUS       sequence-334491-        7705 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid Double Transcriptional Unit - 
            pCmYLCV911_E-Luc_35St+pNOS_Red-F_NOSt.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7705)
  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 7705)
  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: pGREAT25
FEATURES             Location/Qualifiers
     source          1..7705
                     /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        2437..2616
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      4321..4573
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4608..4626)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4815..4839
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4930..5518)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5010..5029)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5689..6480)
                     /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      7124..7559
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    join(7686..7705,1..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 cggccatgct agagtccgca aaaatcacca gtctctctct
     2161 acaaatctat ctctctctat ttttctccag aataatgtgt gagtagttcc cagataaggg
     2221 aattagggtt cttatagggt ttcgctcatg tgttgagcat ataagaaacc cttagtatgt
     2281 atttgtattt gtaaaatact tctatcaata aaatttctaa ttcctaaaac caaaatccag
     2341 tgacctcgct gtcaggagat acatgagaat taagggagtc acggtatgac ccccgccgat
     2401 gacgcgggac aagccgtttt acgtttggaa ctgacagaac cgcaacgttg aaggagccac
     2461 tgagccgcgg gtttctggag tttaatgagc taagcacata cgtcagaaac cattattgcg
     2521 cgttcaaaag tcgcctaagg tcactatcag ctagcaaata tttcttgtca aaaatgctcc
     2581 actgacgttc cataaattcc cctcggtatc caattagagt ctcatattca ctctcaactc
     2641 gatcgaggca atggagaaca tggagaacga cgagaacatt gtggtgggac ccaagccgtt
     2701 ctacccgatc gaggagggtt ccgctggcac ccagctgcgc aagtacatgg agcgctacgc
     2761 taagctgggt gctatcgcct tcaccaacgc cgtgaccggt gtggattact cctacgccga
     2821 gtacctggag aagtcctgct gcctcggaaa ggccctgcag aactacggtc tggtcgtgga
     2881 tggccgcatt gccctctgct cggagaactg cgaggagttc ttcattcccg tgatcgctgg
     2941 tctgttcatc ggagtcggcg ttgccccaac caacgagatc tacaccctgc gcgagctggt
     3001 ccacagcctg ggtatcagca agccgaccat cgttttctcg agcaagaagg gcctcgacaa
     3061 ggttatcacc gtgcaaaaaa ccgtcaccac catcaagacc atcgttatcc tggattccaa
     3121 ggttgactac cgcggttacc agtgcctcga caccttcatc aagcgtaaca ccccacccgg
     3181 cttccaagcc tcctccttca aaaccgtgga ggttgatcgc aaggagcagg tggccctgat
     3241 catgaactcc tccggctcca ccggtctgcc aaagggcgtg cagctgaccc acgagaacac
     3301 cgtcacccgc ttctcgcatg cccgtgatcc catctacggc aaccaggtgt ccccaggcac
     3361 cgccgtcctg accgtcgtgc ccctccacca cggcttcggc atgttcacca ctctcggcta
     3421 cttgatctgc ggcttccgtg tggtgatgct gactaagttc gacgaggaga ctttcctcaa
     3481 gaccctgcaa gactacaagt gcacttacgt gatcctggtg ccgaccctgt tcgccatcct
     3541 gaacaagagc gagctgttga acaaatacga cctgtcgaac ctggtggaga tcgcttccgg
     3601 cggcgcccca ctgtcgaagg aagttggcga ggccgttgct cgccgcttca acctgcccgg
     3661 cgtccgtcag ggttacggcc tgaccgagac tacctcggct atcatcatca cccccgaggg
     3721 cgacgataag cccggcgctt ccggcaaggt tgttccactc ttcaaagcca aggtgatcga
     3781 cctggacacc aagaagtccc tgggtcccaa ccgtcgcgga gaagtgtgcg tcaagggccc
     3841 catgctgatg aagggctacg tcaacaaccc agaggccacc aaggagttga tcgacgagga
     3901 gggctggctg cacactggtg acatcggtta ctacgacgaa gagaagcact tcttcatcgt
     3961 tgaccgcctg aagtccttga tcaagtacaa gggctaccag gttccccccg ccgagctcga
     4021 gagcgtcctg ctgcagcacc cctccatttt cgatgccgga gtggccggtg tccccgatcc
     4081 cgtggctggc gagctgccag gtgccgtcgt cgtgctggag tccggcaaga acatgaccga
     4141 gaaggaagtg atggattacg tggcctcgca agtgtccaac gccaagcgcc tgcgcggcgg
     4201 cgtgcgcttc gtggatgagg tgcccaaggg cctcaccgga aagatcgacg gtcgcgctat
     4261 ccgcgagatc ctcaagaagc ccgtggccaa gatgtgagct tggaatggat cttcgatccc
     4321 gatcgttcaa acatttggca ataaagtttc ttaagattga atcctgttgc cggtcttgcg
     4381 acgattatca tataatttct gttgaattac gttaagcatg taataattaa catgtaatgc
     4441 atgacgttat ttatgagatg ggtttttatg attagagtcc cgcaattata catttaatac
     4501 gcgatagaaa acaaaatata gcgcgcaaac taggataaat tatcgcgcgc ggtgtcatct
     4561 atgttactag atcgggaatt gccaagctaa ttcttgaaga tgaaagggcc tcgtgatacg
     4621 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     4681 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4741 tccgctcatg agacaataac cctgataaat gcttcaataa tgggaccgac tcgcgctgtc
     4801 atgagaccgg atcctgacag gatatattgg cgggtaaact aagtcgctgt atgtgtttgt
     4861 ttgagatctc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4921 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4981 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5041 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5101 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5161 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5221 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5281 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5341 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     5401 gccagttacc ttcggaagaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5461 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5521 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5581 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5641 aagttttaaa tcaatctaaa gtatatatgt gtaacattgg tctagtgatt atttgccgac
     5701 taccttggtg atctcgcctt tcacgtagtg gacaaattct tccaactgat ctgcgcgcga
     5761 ggccaagcga tcttcttctt gtccaagata agcctgtcta gcttcaagta tgacgggctg
     5821 atactgggcc ggcaggcgct ccattgccca gtcggcagcg acatccttcg gcgcgatttt
     5881 gccggttact gcgctgtacc aaatgcggga caacgtaagc actacatttc gctcatcacc
     5941 agcccagtcg ggcggcgagt tccatagcgt taaggtttca tttagcgcct caaatagatc
     6001 ctgttcagga accggatcaa agagttcctc cgccgctgga cctaccaagg caacgctatg
     6061 ttctcttgct tttgtcagca agatagccag atcaatgtcg atcgtggctg gctcgaagat
     6121 acctgcaaga atgtcattgc gctgccattc tccaaattgc agttcgcgct tagctggata
     6181 acgccacgga atgatgtcgt cgtgcacaac aatggtgact tctacagcgc ggagaatctc
     6241 gctctctcca ggggaagccg aagtttccaa aaggtcgttg atcaaagctc gccgcgttgt
     6301 ttcatcaagc cttacggtca ccgtaaccag caaatcaata tcactgtgtg gcttcaggcc
     6361 gccatccact gcggagccgt acaaatgtac ggccagcaac gtcggttcga gatggcgctc
     6421 gatgacgcca actacctctg atagttgagt cgatacttcg gcgatcaccg cttccctcat
     6481 aatgtttaac tttgttttag ggcgactgcc ctgctgcgta acatcgttgc tgctccataa
     6541 catcaaacat cgacccacgg cgtaacgcgc ttgctgcttg gatgcccgag gcatagactg
     6601 taccccaaaa aaacagtcat aacaagccat gaaaaccgcc actgcgccgt taccaccgct
     6661 gcgttcggtc aaggttctgg accagttgcg tgagcgcata cgctacttgc attacagctt
     6721 acgaaccgaa caggcttatg tccactgggt tcgtgccttc atccgtttcc acggtgtgcg
     6781 tcacccggca accttgggta gcagcgaagt cgaggcattt ctgtcctggc tggaacaccc
     6841 cttgtattac tgtttatgta agcagacagt tttattgttc atgatgatat atttttatct
     6901 tgtgcaatgt aacatcagag attttgagac acaacgtggc tttgttgaat aaatcgaact
     6961 tttgctgagt tgaaggatca gatcacgcat cttcccgaca acgcagaccg ttccgtggca
     7021 aagcaaaagt tcaaaatcac caactggtcc acctacaaca aagctctcat caaccgtggc
     7081 tccctcactt tctggctgga tgatggggcg attcaggcga tccccatcca acagcccgcc
     7141 gtcgagcggg cttttttatc cccggaagcc tgtggataga gggtagttat ccacgtgaaa
     7201 ccgctaatgc cccgcaaagc cttgattcac ggggctttcc ggcccgctcc aaaaactatc
     7261 cacgtgaaat cgctaatcag ggtacgtgaa atcgctaatc ggagtacgtg aaatcgctaa
     7321 taaggtcacg tgaaatcgct aatcaaaaag gcacgtgaga acgctaatag ccctttcaga
     7381 tcaacagctt gcaaacaccc ctcgctccgg caagtagtta cagcaagtag tatgttcaat
     7441 tagcttttca attatgaata tatatatcaa ttattggtcg cccttggctt gtggacaatg
     7501 cgctacgcgc accggctccg cccgtggaca accgcaagcg gttgcccacc gtcgagcgcc
     7561 tttgcccaca acccggcggc cggccgcaac agatcgtttt ataaattttt ttttttgaaa
     7621 aagaaaaagc ccgaaaggcg gcaacctctc gggcttctgg atttccgatc cccggaatta
     7681 gatcttggca ggatatattg tggtg
//
