LOCUS       sequence-334488-        7729 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid Double Transcriptional Unit - 
            pCmYLCV911_E-Luc_AtHSP18.2t+pNOS_Red-F_NOSt.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7729)
  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 7729)
  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: pGREAT20
FEATURES             Location/Qualifiers
     source          1..7729
                     /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      2122..2370
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     promoter        2461..2640
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      4345..4597
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4632..4650)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4839..4863
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4954..5542)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5034..5053)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5713..6504)
                     /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      7148..7583
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    join(7710..7729,1..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 atatgaagat gaagatgaaa tatttggtgt gtcaaataaa
     2161 aagcttgtgt gcttaagttt gtgttttttt cttggcttgt tgtgttatga atttgtggct
     2221 ttttctaata ttaaatgaat gtaagatctc attataatga ataaacaaat gtttctataa
     2281 tccattgtga atgttttgtt ggatctcttc tgcagcatat aactactgta tgtgctatgg
     2341 tatggactat ggaatatgat taaagataag cgctgtcagg agatacatga gaattaaggg
     2401 agtcacggta tgacccccgc cgatgacgcg ggacaagccg ttttacgttt ggaactgaca
     2461 gaaccgcaac gttgaaggag ccactgagcc gcgggtttct ggagtttaat gagctaagca
     2521 catacgtcag aaaccattat tgcgcgttca aaagtcgcct aaggtcacta tcagctagca
     2581 aatatttctt gtcaaaaatg ctccactgac gttccataaa ttcccctcgg tatccaatta
     2641 gagtctcata ttcactctca actcgatcga ggcaatggag aacatggaga acgacgagaa
     2701 cattgtggtg ggacccaagc cgttctaccc gatcgaggag ggttccgctg gcacccagct
     2761 gcgcaagtac atggagcgct acgctaagct gggtgctatc gccttcacca acgccgtgac
     2821 cggtgtggat tactcctacg ccgagtacct ggagaagtcc tgctgcctcg gaaaggccct
     2881 gcagaactac ggtctggtcg tggatggccg cattgccctc tgctcggaga actgcgagga
     2941 gttcttcatt cccgtgatcg ctggtctgtt catcggagtc ggcgttgccc caaccaacga
     3001 gatctacacc ctgcgcgagc tggtccacag cctgggtatc agcaagccga ccatcgtttt
     3061 ctcgagcaag aagggcctcg acaaggttat caccgtgcaa aaaaccgtca ccaccatcaa
     3121 gaccatcgtt atcctggatt ccaaggttga ctaccgcggt taccagtgcc tcgacacctt
     3181 catcaagcgt aacaccccac ccggcttcca agcctcctcc ttcaaaaccg tggaggttga
     3241 tcgcaaggag caggtggccc tgatcatgaa ctcctccggc tccaccggtc tgccaaaggg
     3301 cgtgcagctg acccacgaga acaccgtcac ccgcttctcg catgcccgtg atcccatcta
     3361 cggcaaccag gtgtccccag gcaccgccgt cctgaccgtc gtgcccctcc accacggctt
     3421 cggcatgttc accactctcg gctacttgat ctgcggcttc cgtgtggtga tgctgactaa
     3481 gttcgacgag gagactttcc tcaagaccct gcaagactac aagtgcactt acgtgatcct
     3541 ggtgccgacc ctgttcgcca tcctgaacaa gagcgagctg ttgaacaaat acgacctgtc
     3601 gaacctggtg gagatcgctt ccggcggcgc cccactgtcg aaggaagttg gcgaggccgt
     3661 tgctcgccgc ttcaacctgc ccggcgtccg tcagggttac ggcctgaccg agactacctc
     3721 ggctatcatc atcacccccg agggcgacga taagcccggc gcttccggca aggttgttcc
     3781 actcttcaaa gccaaggtga tcgacctgga caccaagaag tccctgggtc ccaaccgtcg
     3841 cggagaagtg tgcgtcaagg gccccatgct gatgaagggc tacgtcaaca acccagaggc
     3901 caccaaggag ttgatcgacg aggagggctg gctgcacact ggtgacatcg gttactacga
     3961 cgaagagaag cacttcttca tcgttgaccg cctgaagtcc ttgatcaagt acaagggcta
     4021 ccaggttccc cccgccgagc tcgagagcgt cctgctgcag cacccctcca ttttcgatgc
     4081 cggagtggcc ggtgtccccg atcccgtggc tggcgagctg ccaggtgccg tcgtcgtgct
     4141 ggagtccggc aagaacatga ccgagaagga agtgatggat tacgtggcct cgcaagtgtc
     4201 caacgccaag cgcctgcgcg gcggcgtgcg cttcgtggat gaggtgccca agggcctcac
     4261 cggaaagatc gacggtcgcg ctatccgcga gatcctcaag aagcccgtgg ccaagatgtg
     4321 agcttggaat ggatcttcga tcccgatcgt tcaaacattt ggcaataaag tttcttaaga
     4381 ttgaatcctg ttgccggtct tgcgacgatt atcatataat ttctgttgaa ttacgttaag
     4441 catgtaataa ttaacatgta atgcatgacg ttatttatga gatgggtttt tatgattaga
     4501 gtcccgcaat tatacattta atacgcgata gaaaacaaaa tatagcgcgc aaactaggat
     4561 aaattatcgc gcgcggtgtc atctatgtta ctagatcggg aattgccaag ctaattcttg
     4621 aagatgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     4681 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4741 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4801 ataatgggac cgactcgcgc tgtcatgaga ccggatcctg acaggatata ttggcgggta
     4861 aactaagtcg ctgtatgtgt ttgtttgaga tctcatgtga gcaaaaggcc agcaaaaggc
     4921 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4981 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5041 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5101 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5161 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5221 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5281 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5341 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5401 atttggtatc tgcgctctgc tgaagccagt taccttcgga agaagagttg gtagctcttg
     5461 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5521 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5581 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5641 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgtgtaaca
     5701 ttggtctagt gattatttgc cgactacctt ggtgatctcg cctttcacgt agtggacaaa
     5761 ttcttccaac tgatctgcgc gcgaggccaa gcgatcttct tcttgtccaa gataagcctg
     5821 tctagcttca agtatgacgg gctgatactg ggccggcagg cgctccattg cccagtcggc
     5881 agcgacatcc ttcggcgcga ttttgccggt tactgcgctg taccaaatgc gggacaacgt
     5941 aagcactaca tttcgctcat caccagccca gtcgggcggc gagttccata gcgttaaggt
     6001 ttcatttagc gcctcaaata gatcctgttc aggaaccgga tcaaagagtt cctccgccgc
     6061 tggacctacc aaggcaacgc tatgttctct tgcttttgtc agcaagatag ccagatcaat
     6121 gtcgatcgtg gctggctcga agatacctgc aagaatgtca ttgcgctgcc attctccaaa
     6181 ttgcagttcg cgcttagctg gataacgcca cggaatgatg tcgtcgtgca caacaatggt
     6241 gacttctaca gcgcggagaa tctcgctctc tccaggggaa gccgaagttt ccaaaaggtc
     6301 gttgatcaaa gctcgccgcg ttgtttcatc aagccttacg gtcaccgtaa ccagcaaatc
     6361 aatatcactg tgtggcttca ggccgccatc cactgcggag ccgtacaaat gtacggccag
     6421 caacgtcggt tcgagatggc gctcgatgac gccaactacc tctgatagtt gagtcgatac
     6481 ttcggcgatc accgcttccc tcataatgtt taactttgtt ttagggcgac tgccctgctg
     6541 cgtaacatcg ttgctgctcc ataacatcaa acatcgaccc acggcgtaac gcgcttgctg
     6601 cttggatgcc cgaggcatag actgtacccc aaaaaaacag tcataacaag ccatgaaaac
     6661 cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt ctggaccagt tgcgtgagcg
     6721 catacgctac ttgcattaca gcttacgaac cgaacaggct tatgtccact gggttcgtgc
     6781 cttcatccgt ttccacggtg tgcgtcaccc ggcaaccttg ggtagcagcg aagtcgaggc
     6841 atttctgtcc tggctggaac accccttgta ttactgttta tgtaagcaga cagttttatt
     6901 gttcatgatg atatattttt atcttgtgca atgtaacatc agagattttg agacacaacg
     6961 tggctttgtt gaataaatcg aacttttgct gagttgaagg atcagatcac gcatcttccc
     7021 gacaacgcag accgttccgt ggcaaagcaa aagttcaaaa tcaccaactg gtccacctac
     7081 aacaaagctc tcatcaaccg tggctccctc actttctggc tggatgatgg ggcgattcag
     7141 gcgatcccca tccaacagcc cgccgtcgag cgggcttttt tatccccgga agcctgtgga
     7201 tagagggtag ttatccacgt gaaaccgcta atgccccgca aagccttgat tcacggggct
     7261 ttccggcccg ctccaaaaac tatccacgtg aaatcgctaa tcagggtacg tgaaatcgct
     7321 aatcggagta cgtgaaatcg ctaataaggt cacgtgaaat cgctaatcaa aaaggcacgt
     7381 gagaacgcta atagcccttt cagatcaaca gcttgcaaac acccctcgct ccggcaagta
     7441 gttacagcaa gtagtatgtt caattagctt ttcaattatg aatatatata tcaattattg
     7501 gtcgcccttg gcttgtggac aatgcgctac gcgcaccggc tccgcccgtg gacaaccgca
     7561 agcggttgcc caccgtcgag cgcctttgcc cacaacccgg cggccggccg caacagatcg
     7621 ttttataaat tttttttttt gaaaaagaaa aagcccgaaa ggcggcaacc tctcgggctt
     7681 ctggatttcc gatccccgga attagatctt ggcaggatat attgtggtg
//
