LOCUS       sequence-335142-        7971 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid Double Transcriptional Unit - 
            pCmYLCV911_E-Luc_NOSt+pNOS_Red-F_NOSt.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7971)
  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 7971)
  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: pGREAT5
FEATURES             Location/Qualifiers
     source          1..7971
                     /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)"
     terminator      2140..2392
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        2703..2882
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      4587..4839
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    5081..5105
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5196..5784)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5276..5295)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5955..6746)
                     /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      7390..7825
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    join(7952..7971,1..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 ggaatggatc ttcgatcccg atcgttcaaa catttggcaa
     2161 taaagtttct taagattgaa tcctgttgcc ggtcttgcga cgattatcat ataatttctg
     2221 ttgaattacg ttaagcatgt aataattaac atgtaatgca tgacgttatt tatgagatgg
     2281 gtttttatga ttagagtccc gcaattatac atttaatacg cgatagaaaa caaaatatag
     2341 cgcgcaaact aggataaatt atcgcgcgcg gtgtcatcta tgttactaga tcgggaattg
     2401 ccaagctaat tcttgaagat gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     2461 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     2521 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     2581 ctgataaatg cttcaataat gggaccgact cgcgctgtca ggagatacat gagaattaag
     2641 ggagtcacgg tatgaccccc gccgatgacg cgggacaagc cgttttacgt ttggaactga
     2701 cagaaccgca acgttgaagg agccactgag ccgcgggttt ctggagttta atgagctaag
     2761 cacatacgtc agaaaccatt attgcgcgtt caaaagtcgc ctaaggtcac tatcagctag
     2821 caaatatttc ttgtcaaaaa tgctccactg acgttccata aattcccctc ggtatccaat
     2881 tagagtctca tattcactct caactcgatc gaggcaatgg agaacatgga gaacgacgag
     2941 aacattgtgg tgggacccaa gccgttctac ccgatcgagg agggttccgc tggcacccag
     3001 ctgcgcaagt acatggagcg ctacgctaag ctgggtgcta tcgccttcac caacgccgtg
     3061 accggtgtgg attactccta cgccgagtac ctggagaagt cctgctgcct cggaaaggcc
     3121 ctgcagaact acggtctggt cgtggatggc cgcattgccc tctgctcgga gaactgcgag
     3181 gagttcttca ttcccgtgat cgctggtctg ttcatcggag tcggcgttgc cccaaccaac
     3241 gagatctaca ccctgcgcga gctggtccac agcctgggta tcagcaagcc gaccatcgtt
     3301 ttctcgagca agaagggcct cgacaaggtt atcaccgtgc aaaaaaccgt caccaccatc
     3361 aagaccatcg ttatcctgga ttccaaggtt gactaccgcg gttaccagtg cctcgacacc
     3421 ttcatcaagc gtaacacccc acccggcttc caagcctcct ccttcaaaac cgtggaggtt
     3481 gatcgcaagg agcaggtggc cctgatcatg aactcctccg gctccaccgg tctgccaaag
     3541 ggcgtgcagc tgacccacga gaacaccgtc acccgcttct cgcatgcccg tgatcccatc
     3601 tacggcaacc aggtgtcccc aggcaccgcc gtcctgaccg tcgtgcccct ccaccacggc
     3661 ttcggcatgt tcaccactct cggctacttg atctgcggct tccgtgtggt gatgctgact
     3721 aagttcgacg aggagacttt cctcaagacc ctgcaagact acaagtgcac ttacgtgatc
     3781 ctggtgccga ccctgttcgc catcctgaac aagagcgagc tgttgaacaa atacgacctg
     3841 tcgaacctgg tggagatcgc ttccggcggc gccccactgt cgaaggaagt tggcgaggcc
     3901 gttgctcgcc gcttcaacct gcccggcgtc cgtcagggtt acggcctgac cgagactacc
     3961 tcggctatca tcatcacccc cgagggcgac gataagcccg gcgcttccgg caaggttgtt
     4021 ccactcttca aagccaaggt gatcgacctg gacaccaaga agtccctggg tcccaaccgt
     4081 cgcggagaag tgtgcgtcaa gggccccatg ctgatgaagg gctacgtcaa caacccagag
     4141 gccaccaagg agttgatcga cgaggagggc tggctgcaca ctggtgacat cggttactac
     4201 gacgaagaga agcacttctt catcgttgac cgcctgaagt ccttgatcaa gtacaagggc
     4261 taccaggttc cccccgccga gctcgagagc gtcctgctgc agcacccctc cattttcgat
     4321 gccggagtgg ccggtgtccc cgatcccgtg gctggcgagc tgccaggtgc cgtcgtcgtg
     4381 ctggagtccg gcaagaacat gaccgagaag gaagtgatgg attacgtggc ctcgcaagtg
     4441 tccaacgcca agcgcctgcg cggcggcgtg cgcttcgtgg atgaggtgcc caagggcctc
     4501 accggaaaga tcgacggtcg cgctatccgc gagatcctca agaagcccgt ggccaagatg
     4561 tgagcttgga atggatcttc gatcccgatc gttcaaacat ttggcaataa agtttcttaa
     4621 gattgaatcc tgttgccggt cttgcgacga ttatcatata atttctgttg aattacgtta
     4681 agcatgtaat aattaacatg taatgcatga cgttatttat gagatgggtt tttatgatta
     4741 gagtcccgca attatacatt taatacgcga tagaaaacaa aatatagcgc gcaaactagg
     4801 ataaattatc gcgcgcggtg tcatctatgt tactagatcg ggaattgcca agctaattct
     4861 tgaagatgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     4921 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4981 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5041 caataatggg accgactcgc gctgtcatga gaccggatcc tgacaggata tattggcggg
     5101 taaactaagt cgctgtatgt gtttgtttga gatctcatgt gagcaaaagg ccagcaaaag
     5161 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5221 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5281 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5341 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5401 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5461 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5521 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5581 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5641 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaagaagagt tggtagctct
     5701 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5761 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5821 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5881 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgtgtaa
     5941 cattggtcta gtgattattt gccgactacc ttggtgatct cgcctttcac gtagtggaca
     6001 aattcttcca actgatctgc gcgcgaggcc aagcgatctt cttcttgtcc aagataagcc
     6061 tgtctagctt caagtatgac gggctgatac tgggccggca ggcgctccat tgcccagtcg
     6121 gcagcgacat ccttcggcgc gattttgccg gttactgcgc tgtaccaaat gcgggacaac
     6181 gtaagcacta catttcgctc atcaccagcc cagtcgggcg gcgagttcca tagcgttaag
     6241 gtttcattta gcgcctcaaa tagatcctgt tcaggaaccg gatcaaagag ttcctccgcc
     6301 gctggaccta ccaaggcaac gctatgttct cttgcttttg tcagcaagat agccagatca
     6361 atgtcgatcg tggctggctc gaagatacct gcaagaatgt cattgcgctg ccattctcca
     6421 aattgcagtt cgcgcttagc tggataacgc cacggaatga tgtcgtcgtg cacaacaatg
     6481 gtgacttcta cagcgcggag aatctcgctc tctccagggg aagccgaagt ttccaaaagg
     6541 tcgttgatca aagctcgccg cgttgtttca tcaagcctta cggtcaccgt aaccagcaaa
     6601 tcaatatcac tgtgtggctt caggccgcca tccactgcgg agccgtacaa atgtacggcc
     6661 agcaacgtcg gttcgagatg gcgctcgatg acgccaacta cctctgatag ttgagtcgat
     6721 acttcggcga tcaccgcttc cctcataatg tttaactttg ttttagggcg actgccctgc
     6781 tgcgtaacat cgttgctgct ccataacatc aaacatcgac ccacggcgta acgcgcttgc
     6841 tgcttggatg cccgaggcat agactgtacc ccaaaaaaac agtcataaca agccatgaaa
     6901 accgccactg cgccgttacc accgctgcgt tcggtcaagg ttctggacca gttgcgtgag
     6961 cgcatacgct acttgcatta cagcttacga accgaacagg cttatgtcca ctgggttcgt
     7021 gccttcatcc gtttccacgg tgtgcgtcac ccggcaacct tgggtagcag cgaagtcgag
     7081 gcatttctgt cctggctgga acaccccttg tattactgtt tatgtaagca gacagtttta
     7141 ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt tgagacacaa
     7201 cgtggctttg ttgaataaat cgaacttttg ctgagttgaa ggatcagatc acgcatcttc
     7261 ccgacaacgc agaccgttcc gtggcaaagc aaaagttcaa aatcaccaac tggtccacct
     7321 acaacaaagc tctcatcaac cgtggctccc tcactttctg gctggatgat ggggcgattc
     7381 aggcgatccc catccaacag cccgccgtcg agcgggcttt tttatccccg gaagcctgtg
     7441 gatagagggt agttatccac gtgaaaccgc taatgccccg caaagccttg attcacgggg
     7501 ctttccggcc cgctccaaaa actatccacg tgaaatcgct aatcagggta cgtgaaatcg
     7561 ctaatcggag tacgtgaaat cgctaataag gtcacgtgaa atcgctaatc aaaaaggcac
     7621 gtgagaacgc taatagccct ttcagatcaa cagcttgcaa acacccctcg ctccggcaag
     7681 tagttacagc aagtagtatg ttcaattagc ttttcaatta tgaatatata tatcaattat
     7741 tggtcgccct tggcttgtgg acaatgcgct acgcgcaccg gctccgcccg tggacaaccg
     7801 caagcggttg cccaccgtcg agcgcctttg cccacaaccc ggcggccggc cgcaacagat
     7861 cgttttataa attttttttt ttgaaaaaga aaaagcccga aaggcggcaa cctctcgggc
     7921 ttctggattt ccgatccccg gaattagatc ttggcaggat atattgtggt g
//
