LOCUS       sequence-334422-        7629 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid Double Transcriptional Unit - 
            pAtNR1_E-Luc_AtHSP18.2t+pNOS_Red-F_NOSt.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7629)
  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 7629)
  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: pGREAT27
FEATURES             Location/Qualifiers
     source          1..7629
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     63..82
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    253..277
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     466..484
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(519..771)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(2476..2655)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      complement(2746..2994)
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     RBS             3148..3156
                     /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        complement(4631..4677)
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     misc_feature    5013..5035
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     rep_origin      5162..5597
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     CDS             6241..7032
                     /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      join(7203..7629,1..162)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
       61 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
      121 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
      181 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgagatctca aacaaacaca
      241 tacagcgact tagtttaccc gccaatatat cctgtcagga tccggtctca tgacagcgcg
      301 agtcggtccc attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
      361 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
      421 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
      481 aggccctttc atcttcaaga attagcttgg caattcccga tctagtaaca tagatgacac
      541 cgcgcgcgat aatttatcct agtttgcgcg ctatattttg ttttctatcg cgtattaaat
      601 gtataattgc gggactctaa tcataaaaac ccatctcata aataacgtca tgcattacat
      661 gttaattatt acatgcttaa cgtaattcaa cagaaattat atgataatcg tcgcaagacc
      721 ggcaacagga ttcaatctta agaaacttta ttgccaaatg tttgaacgat cgggatcgaa
      781 gatccattcc aagctcacat cttggccacg ggcttcttga ggatctcgcg gatagcgcga
      841 ccgtcgatct ttccggtgag gcccttgggc acctcatcca cgaagcgcac gccgccgcgc
      901 aggcgcttgg cgttggacac ttgcgaggcc acgtaatcca tcacttcctt ctcggtcatg
      961 ttcttgccgg actccagcac gacgacggca cctggcagct cgccagccac gggatcgggg
     1021 acaccggcca ctccggcatc gaaaatggag gggtgctgca gcaggacgct ctcgagctcg
     1081 gcggggggaa cctggtagcc cttgtacttg atcaaggact tcaggcggtc aacgatgaag
     1141 aagtgcttct cttcgtcgta gtaaccgatg tcaccagtgt gcagccagcc ctcctcgtcg
     1201 atcaactcct tggtggcctc tgggttgttg acgtagccct tcatcagcat ggggcccttg
     1261 acgcacactt ctccgcgacg gttgggaccc agggacttct tggtgtccag gtcgatcacc
     1321 ttggctttga agagtggaac aaccttgccg gaagcgccgg gcttatcgtc gccctcgggg
     1381 gtgatgatga tagccgaggt agtctcggtc aggccgtaac cctgacggac gccgggcagg
     1441 ttgaagcggc gagcaacggc ctcgccaact tccttcgaca gtggggcgcc gccggaagcg
     1501 atctccacca ggttcgacag gtcgtatttg ttcaacagct cgctcttgtt caggatggcg
     1561 aacagggtcg gcaccaggat cacgtaagtg cacttgtagt cttgcagggt cttgaggaaa
     1621 gtctcctcgt cgaacttagt cagcatcacc acacggaagc cgcagatcaa gtagccgaga
     1681 gtggtgaaca tgccgaagcc gtggtggagg ggcacgacgg tcaggacggc ggtgcctggg
     1741 gacacctggt tgccgtagat gggatcacgg gcatgcgaga agcgggtgac ggtgttctcg
     1801 tgggtcagct gcacgccctt tggcagaccg gtggagccgg aggagttcat gatcagggcc
     1861 acctgctcct tgcgatcaac ctccacggtt ttgaaggagg aggcttggaa gccgggtggg
     1921 gtgttacgct tgatgaaggt gtcgaggcac tggtaaccgc ggtagtcaac cttggaatcc
     1981 aggataacga tggtcttgat ggtggtgacg gttttttgca cggtgataac cttgtcgagg
     2041 cccttcttgc tcgagaaaac gatggtcggc ttgctgatac ccaggctgtg gaccagctcg
     2101 cgcagggtgt agatctcgtt ggttggggca acgccgactc cgatgaacag accagcgatc
     2161 acgggaatga agaactcctc gcagttctcc gagcagaggg caatgcggcc atccacgacc
     2221 agaccgtagt tctgcagggc ctttccgagg cagcaggact tctccaggta ctcggcgtag
     2281 gagtaatcca caccggtcac ggcgttggtg aaggcgatag cacccagctt agcgtagcgc
     2341 tccatgtact tgcgcagctg ggtgccagcg gaaccctcct cgatcgggta gaacggcttg
     2401 ggtcccacca caatgttctc gtcgttctcc atgttctcca ttgcctcgat cgagttgaga
     2461 gtgaatatga gactctaatt ggataccgag gggaatttat ggaacgtcag tggagcattt
     2521 ttgacaagaa atatttgcta gctgatagtg accttaggcg acttttgaac gcgcaataat
     2581 ggtttctgac gtatgtgctt agctcattaa actccagaaa cccgcggctc agtggctcct
     2641 tcaacgttgc ggttctgtca gttccaaacg taaaacggct tgtcccgcgt catcggcggg
     2701 ggtcataccg tgactccctt aattctcatg tatctcctga cagcgcttat ctttaatcat
     2761 attccatagt ccataccata gcacatacag tagttatatg ctgcagaaga gatccaacaa
     2821 aacattcaca atggattata gaaacatttg tttattcatt ataatgagat cttacattca
     2881 tttaatatta gaaaaagcca caaattcata acacaacaag ccaagaaaaa aacacaaact
     2941 taagcacaca agctttttat ttgacacacc aaatatttca tcttcatctt catataagct
     3001 tacagcttag aagccttctc catcagggcc tccctcagct cttttcgact aatcttgcct
     3061 gtcacgttcc tggggataga gtccacgaac cttacgccgc ccctcaggta cttggagtga
     3121 gacaccctct gggccaggaa gtcgtacacc tccttagcgg tgagttgggc gccgggctgc
     3181 ttcaccacga agccggctgg cagctcgccg gcctcgatgt cagggatacc cacgacggcc
     3241 acgtccctga tgcctgggtg ctgaaggagg atctcctcca gctccacagg ggcgacctga
     3301 tagcccttgt atttgatcag ctccttgtac cggtccacga tgtagaaata ctcgtcctcg
     3361 tcgtagtagc cgaagtcgcc agagtgcagc cagccgtcgt cgtcgatggc ctccttagta
     3421 gcctgtgggt tgttcacata gccttttgtt accataggtc cccagatgca cagctcgccc
     3481 acctggtttg gacccagggc ctcgccggtg ttcctgtcga tgatcttggc ggccatgtaa
     3541 ggtgtcacct tgcccaggga gccggacttg aactcgttgt gcagagtatg gatgttggcg
     3601 gaggtagact ctgttagacc gtagccacac cgtatccctg gcaggttcag cctcttcacg
     3661 gcgatctcgg ccacctcctt cgccagagga gcggctccac agcacagctc cgccagggtg
     3721 gataggtcgt acttgtccac cagagggctc ttggacagga acaggattgt ggagggaacg
     3781 ttgatcacgc tcctcacctc gtagtcctgg atggccttca ggaacacctc ctggttaaac
     3841 cttcggagca tcaccactct caggcccacc atgaaatagc ccaggttgat actgaagccg
     3901 aaggcgtgga agaatggcag gtaggccagc acggatacgc cggggatgag ttgtgtaccc
     3961 actctggggt cagatgcgtg tgtgagtctc acacagatgt tcctgtgggt ctgcatcacg
     4021 cctttaggca ggccggttgt gccggaggag cacaggatgg cggctacctg gtccacaggg
     4081 tcgtagtgca gaggcttgaa tgtttggagg ttgttgtcgg agtacctgga catgaagttg
     4141 tgcaggctct cgcagcccag caggttctct tcagagtcca gtatgatgat cctctttatg
     4201 tagttggttc tgtctttaac ctccaaaacc ttaggcagga tcttcctagt ggtaaagacc
     4261 aggatcggct tggagatgcc ggtcacttta cacagctcgt ctgggatata gtcctcgttc
     4321 acaggggcca ccaccatgcc gatgtaccag gcggagatga tggggatgaa gaatctcttg
     4381 ttgttctctg cacagatcga cacgacatcg ttcatcttgt agccacagtt ctggaggctc
     4441 tggcccagct tcacagtagt gtcgaagaac tcctggtagg aaagggactc gttgcccatc
     4501 acgtccagga tggcctgggg gatgtgggag tgcttgtgca gagcgttgta cagcatctcg
     4561 ccggcggtga agttgcccag agggtgcttg ggctcggggc cgtacaccac gttcttctct
     4621 ctctccattt tcctctccaa atgaaatgaa cttccttata tagaggaagg gtcttgcagc
     4681 tcttgagcaa tgtaaagggt caagttgttt ctctttgatc gtatgttgcg aggaggaaac
     4741 agacaccttc tgatttctat ttgttttatt ctttctaaat ttatttgaaa ctatttaaat
     4801 tgtgtttact gcaggataaa gcgcaattaa gtataaacta aacagagatt atggtataaa
     4861 aaaggagttt agagagaaga ctttggaggc taagtggcgc atgggaacac caacggccaa
     4921 gcaaatacac gaacggggca accaaacaaa gggacgggtc gtaatattat ttttgtcact
     4981 tgtttgaatt ttcgctcctg agaccggatc cgttacacca caatatatcc tgccaagatc
     5041 taattccggg gatcggaaat ccagaagccc gagaggttgc cgcctttcgg gctttttctt
     5101 tttcaaaaaa aaaaatttat aaaacgatct gttgcggccg gccgccgggt tgtgggcaaa
     5161 ggcgctcgac ggtgggcaac cgcttgcggt tgtccacggg cggagccggt gcgcgtagcg
     5221 cattgtccac aagccaaggg cgaccaataa ttgatatata tattcataat tgaaaagcta
     5281 attgaacata ctacttgctg taactacttg ccggagcgag gggtgtttgc aagctgttga
     5341 tctgaaaggg ctattagcgt tctcacgtgc ctttttgatt agcgatttca cgtgacctta
     5401 ttagcgattt cacgtactcc gattagcgat ttcacgtacc ctgattagcg atttcacgtg
     5461 gatagttttt ggagcgggcc ggaaagcccc gtgaatcaag gctttgcggg gcattagcgg
     5521 tttcacgtgg ataactaccc tctatccaca ggcttccggg gataaaaaag cccgctcgac
     5581 ggcgggctgt tggatgggga tcgcctgaat cgccccatca tccagccaga aagtgaggga
     5641 gccacggttg atgagagctt tgttgtaggt ggaccagttg gtgattttga acttttgctt
     5701 tgccacggaa cggtctgcgt tgtcgggaag atgcgtgatc tgatccttca actcagcaaa
     5761 agttcgattt attcaacaaa gccacgttgt gtctcaaaat ctctgatgtt acattgcaca
     5821 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     5881 gggtgttcca gccaggacag aaatgcctcg acttcgctgc tacccaaggt tgccgggtga
     5941 cgcacaccgt ggaaacggat gaaggcacga acccagtgga cataagcctg ttcggttcgt
     6001 aagctgtaat gcaagtagcg tatgcgctca cgcaactggt ccagaacctt gaccgaacgc
     6061 agcggtggta acggcgcagt ggcggttttc atggcttgtt atgactgttt ttttggggta
     6121 cagtctatgc ctcgggcatc caagcagcaa gcgcgttacg ccgtgggtcg atgtttgatg
     6181 ttatggagca gcaacgatgt tacgcagcag ggcagtcgcc ctaaaacaaa gttaaacatt
     6241 atgagggaag cggtgatcgc cgaagtatcg actcaactat cagaggtagt tggcgtcatc
     6301 gagcgccatc tcgaaccgac gttgctggcc gtacatttgt acggctccgc agtggatggc
     6361 ggcctgaagc cacacagtga tattgatttg ctggttacgg tgaccgtaag gcttgatgaa
     6421 acaacgcggc gagctttgat caacgacctt ttggaaactt cggcttcccc tggagagagc
     6481 gagattctcc gcgctgtaga agtcaccatt gttgtgcacg acgacatcat tccgtggcgt
     6541 tatccagcta agcgcgaact gcaatttgga gaatggcagc gcaatgacat tcttgcaggt
     6601 atcttcgagc cagccacgat cgacattgat ctggctatct tgctgacaaa agcaagagaa
     6661 catagcgttg ccttggtagg tccagcggcg gaggaactct ttgatccggt tcctgaacag
     6721 gatctatttg aggcgctaaa tgaaacctta acgctatgga actcgccgcc cgactgggct
     6781 ggtgatgagc gaaatgtagt gcttacgttg tcccgcattt ggtacagcgc agtaaccggc
     6841 aaaatcgcgc cgaaggatgt cgctgccgac tgggcaatgg agcgcctgcc ggcccagtat
     6901 cagcccgtca tacttgaagc tagacaggct tatcttggac aagaagaaga tcgcttggcc
     6961 tcgcgcgcag atcagttgga agaatttgtc cactacgtga aaggcgagat caccaaggta
     7021 gtcggcaaat aatcactaga ccaatgttac acatatatac tttagattga tttaaaactt
     7081 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     7141 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     7201 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     7261 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc ttcttccgaa ggtaactggc
     7321 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     7381 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     7441 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     7501 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     7561 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     7621 gggagaaag
//
