LOCUS       sequence-334475-        7702 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid Double Transcriptional Unit - 
            p35SoTMV_E-Luc_AtHSP18.2t+pNOS_Red-F_NOSt.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7702)
  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 7702)
  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: pGREAT17
FEATURES             Location/Qualifiers
     source          1..7702
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        22..367
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     310..330
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    387..442
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     RBS             1933..1941
                     /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      2095..2343
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     promoter        2434..2613
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      4318..4570
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4605..4623)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4812..4836
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4927..5515)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5007..5026)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5686..6477)
                     /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      7121..7556
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    join(7683..7702,1..3)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
ORIGIN
        1 taacggatcc ggtctcagga gtgagacttt tcaacaaagg gtaatatcgg gaaacctcct
       61 cggattccat tgcccagcta tctgtcactt catcgaaagg acagtagaaa aggaaggtgg
      121 ctcctacaaa tgccatcatt gcgataaagg aaaggctatc gttcaagatg cctctaccga
      181 cagtggtccc aaagatggac ccccacccac gaggaacatc gtggaaaaag aagatgttcc
      241 aaccacgtcc tcaaagcaag tggattgatg tgatatctcc actgacgtaa gggatgacgc
      301 acaatcccac tatccttcgc aagacccttc ctctatataa ggaagttcat ttcatttgga
      361 gaggacaggc ttcttgagat ccttcaacaa ttaccaacaa caacaaacaa caaacaacat
      421 tacaattact atttacaatt acagtcgact ctagaggata atggagagag agaagaacgt
      481 ggtgtacggc cccgagccca agcaccctct gggcaacttc accgccggcg agatgctgta
      541 caacgctctg cacaagcact cccacatccc ccaggccatc ctggacgtga tgggcaacga
      601 gtccctttcc taccaggagt tcttcgacac tactgtgaag ctgggccaga gcctccagaa
      661 ctgtggctac aagatgaacg atgtcgtgtc gatctgtgca gagaacaaca agagattctt
      721 catccccatc atctccgcct ggtacatcgg catggtggtg gcccctgtga acgaggacta
      781 tatcccagac gagctgtgta aagtgaccgg catctccaag ccgatcctgg tctttaccac
      841 taggaagatc ctgcctaagg ttttggaggt taaagacaga accaactaca taaagaggat
      901 catcatactg gactctgaag agaacctgct gggctgcgag agcctgcaca acttcatgtc
      961 caggtactcc gacaacaacc tccaaacatt caagcctctg cactacgacc ctgtggacca
     1021 ggtagccgcc atcctgtgct cctccggcac aaccggcctg cctaaaggcg tgatgcagac
     1081 ccacaggaac atctgtgtga gactcacaca cgcatctgac cccagagtgg gtacacaact
     1141 catccccggc gtatccgtgc tggcctacct gccattcttc cacgccttcg gcttcagtat
     1201 caacctgggc tatttcatgg tgggcctgag agtggtgatg ctccgaaggt ttaaccagga
     1261 ggtgttcctg aaggccatcc aggactacga ggtgaggagc gtgatcaacg ttccctccac
     1321 aatcctgttc ctgtccaaga gccctctggt ggacaagtac gacctatcca ccctggcgga
     1381 gctgtgctgt ggagccgctc ctctggcgaa ggaggtggcc gagatcgccg tgaagaggct
     1441 gaacctgcca gggatacggt gtggctacgg tctaacagag tctacctccg ccaacatcca
     1501 tactctgcac aacgagttca agtccggctc cctgggcaag gtgacacctt acatggccgc
     1561 caagatcatc gacaggaaca ccggcgaggc cctgggtcca aaccaggtgg gcgagctgtg
     1621 catctgggga cctatggtaa caaaaggcta tgtgaacaac ccacaggcta ctaaggaggc
     1681 catcgacgac gacggctggc tgcactctgg cgacttcggc tactacgacg aggacgagta
     1741 tttctacatc gtggaccggt acaaggagct gatcaaatac aagggctatc aggtcgcccc
     1801 tgtggagctg gaggagatcc tccttcagca cccaggcatc agggacgtgg ccgtcgtggg
     1861 tatccctgac atcgaggccg gcgagctgcc agccggcttc gtggtgaagc agcccggcgc
     1921 ccaactcacc gctaaggagg tgtacgactt cctggcccag agggtgtctc actccaagta
     1981 cctgaggggc ggcgtaaggt tcgtggactc tatccccagg aacgtgacag gcaagattag
     2041 tcgaaaagag ctgagggagg ccctgatgga gaaggcttct aagctgtaag cttatatgaa
     2101 gatgaagatg aaatatttgg tgtgtcaaat aaaaagcttg tgtgcttaag tttgtgtttt
     2161 tttcttggct tgttgtgtta tgaatttgtg gctttttcta atattaaatg aatgtaagat
     2221 ctcattataa tgaataaaca aatgtttcta taatccattg tgaatgtttt gttggatctc
     2281 ttctgcagca tataactact gtatgtgcta tggtatggac tatggaatat gattaaagat
     2341 aagcgctgtc aggagataca tgagaattaa gggagtcacg gtatgacccc cgccgatgac
     2401 gcgggacaag ccgttttacg tttggaactg acagaaccgc aacgttgaag gagccactga
     2461 gccgcgggtt tctggagttt aatgagctaa gcacatacgt cagaaaccat tattgcgcgt
     2521 tcaaaagtcg cctaaggtca ctatcagcta gcaaatattt cttgtcaaaa atgctccact
     2581 gacgttccat aaattcccct cggtatccaa ttagagtctc atattcactc tcaactcgat
     2641 cgaggcaatg gagaacatgg agaacgacga gaacattgtg gtgggaccca agccgttcta
     2701 cccgatcgag gagggttccg ctggcaccca gctgcgcaag tacatggagc gctacgctaa
     2761 gctgggtgct atcgccttca ccaacgccgt gaccggtgtg gattactcct acgccgagta
     2821 cctggagaag tcctgctgcc tcggaaaggc cctgcagaac tacggtctgg tcgtggatgg
     2881 ccgcattgcc ctctgctcgg agaactgcga ggagttcttc attcccgtga tcgctggtct
     2941 gttcatcgga gtcggcgttg ccccaaccaa cgagatctac accctgcgcg agctggtcca
     3001 cagcctgggt atcagcaagc cgaccatcgt tttctcgagc aagaagggcc tcgacaaggt
     3061 tatcaccgtg caaaaaaccg tcaccaccat caagaccatc gttatcctgg attccaaggt
     3121 tgactaccgc ggttaccagt gcctcgacac cttcatcaag cgtaacaccc cacccggctt
     3181 ccaagcctcc tccttcaaaa ccgtggaggt tgatcgcaag gagcaggtgg ccctgatcat
     3241 gaactcctcc ggctccaccg gtctgccaaa gggcgtgcag ctgacccacg agaacaccgt
     3301 cacccgcttc tcgcatgccc gtgatcccat ctacggcaac caggtgtccc caggcaccgc
     3361 cgtcctgacc gtcgtgcccc tccaccacgg cttcggcatg ttcaccactc tcggctactt
     3421 gatctgcggc ttccgtgtgg tgatgctgac taagttcgac gaggagactt tcctcaagac
     3481 cctgcaagac tacaagtgca cttacgtgat cctggtgccg accctgttcg ccatcctgaa
     3541 caagagcgag ctgttgaaca aatacgacct gtcgaacctg gtggagatcg cttccggcgg
     3601 cgccccactg tcgaaggaag ttggcgaggc cgttgctcgc cgcttcaacc tgcccggcgt
     3661 ccgtcagggt tacggcctga ccgagactac ctcggctatc atcatcaccc ccgagggcga
     3721 cgataagccc ggcgcttccg gcaaggttgt tccactcttc aaagccaagg tgatcgacct
     3781 ggacaccaag aagtccctgg gtcccaaccg tcgcggagaa gtgtgcgtca agggccccat
     3841 gctgatgaag ggctacgtca acaacccaga ggccaccaag gagttgatcg acgaggaggg
     3901 ctggctgcac actggtgaca tcggttacta cgacgaagag aagcacttct tcatcgttga
     3961 ccgcctgaag tccttgatca agtacaaggg ctaccaggtt ccccccgccg agctcgagag
     4021 cgtcctgctg cagcacccct ccattttcga tgccggagtg gccggtgtcc ccgatcccgt
     4081 ggctggcgag ctgccaggtg ccgtcgtcgt gctggagtcc ggcaagaaca tgaccgagaa
     4141 ggaagtgatg gattacgtgg cctcgcaagt gtccaacgcc aagcgcctgc gcggcggcgt
     4201 gcgcttcgtg gatgaggtgc ccaagggcct caccggaaag atcgacggtc gcgctatccg
     4261 cgagatcctc aagaagcccg tggccaagat gtgagcttgg aatggatctt cgatcccgat
     4321 cgttcaaaca tttggcaata aagtttctta agattgaatc ctgttgccgg tcttgcgacg
     4381 attatcatat aatttctgtt gaattacgtt aagcatgtaa taattaacat gtaatgcatg
     4441 acgttattta tgagatgggt ttttatgatt agagtcccgc aattatacat ttaatacgcg
     4501 atagaaaaca aaatatagcg cgcaaactag gataaattat cgcgcgcggt gtcatctatg
     4561 ttactagatc gggaattgcc aagctaattc ttgaagatga aagggcctcg tgatacgcct
     4621 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     4681 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4741 gctcatgaga caataaccct gataaatgct tcaataatgg gaccgactcg cgctgtcatg
     4801 agaccggatc ctgacaggat atattggcgg gtaaactaag tcgctgtatg tgtttgtttg
     4861 agatctcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4921 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4981 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5041 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5101 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5161 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5221 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5281 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5341 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     5401 agttaccttc ggaagaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5461 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5521 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5581 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5641 ttttaaatca atctaaagta tatatgtgta acattggtct agtgattatt tgccgactac
     5701 cttggtgatc tcgcctttca cgtagtggac aaattcttcc aactgatctg cgcgcgaggc
     5761 caagcgatct tcttcttgtc caagataagc ctgtctagct tcaagtatga cgggctgata
     5821 ctgggccggc aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc
     5881 ggttactgcg ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct catcaccagc
     5941 ccagtcgggc ggcgagttcc atagcgttaa ggtttcattt agcgcctcaa atagatcctg
     6001 ttcaggaacc ggatcaaaga gttcctccgc cgctggacct accaaggcaa cgctatgttc
     6061 tcttgctttt gtcagcaaga tagccagatc aatgtcgatc gtggctggct cgaagatacc
     6121 tgcaagaatg tcattgcgct gccattctcc aaattgcagt tcgcgcttag ctggataacg
     6181 ccacggaatg atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga gaatctcgct
     6241 ctctccaggg gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc
     6301 atcaagcctt acggtcaccg taaccagcaa atcaatatca ctgtgtggct tcaggccgcc
     6361 atccactgcg gagccgtaca aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat
     6421 gacgccaact acctctgata gttgagtcga tacttcggcg atcaccgctt ccctcataat
     6481 gtttaacttt gttttagggc gactgccctg ctgcgtaaca tcgttgctgc tccataacat
     6541 caaacatcga cccacggcgt aacgcgcttg ctgcttggat gcccgaggca tagactgtac
     6601 cccaaaaaaa cagtcataac aagccatgaa aaccgccact gcgccgttac caccgctgcg
     6661 ttcggtcaag gttctggacc agttgcgtga gcgcatacgc tacttgcatt acagcttacg
     6721 aaccgaacag gcttatgtcc actgggttcg tgccttcatc cgtttccacg gtgtgcgtca
     6781 cccggcaacc ttgggtagca gcgaagtcga ggcatttctg tcctggctgg aacacccctt
     6841 gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt
     6901 gcaatgtaac atcagagatt ttgagacaca acgtggcttt gttgaataaa tcgaactttt
     6961 gctgagttga aggatcagat cacgcatctt cccgacaacg cagaccgttc cgtggcaaag
     7021 caaaagttca aaatcaccaa ctggtccacc tacaacaaag ctctcatcaa ccgtggctcc
     7081 ctcactttct ggctggatga tggggcgatt caggcgatcc ccatccaaca gcccgccgtc
     7141 gagcgggctt ttttatcccc ggaagcctgt ggatagaggg tagttatcca cgtgaaaccg
     7201 ctaatgcccc gcaaagcctt gattcacggg gctttccggc ccgctccaaa aactatccac
     7261 gtgaaatcgc taatcagggt acgtgaaatc gctaatcgga gtacgtgaaa tcgctaataa
     7321 ggtcacgtga aatcgctaat caaaaaggca cgtgagaacg ctaatagccc tttcagatca
     7381 acagcttgca aacacccctc gctccggcaa gtagttacag caagtagtat gttcaattag
     7441 cttttcaatt atgaatatat atatcaatta ttggtcgccc ttggcttgtg gacaatgcgc
     7501 tacgcgcacc ggctccgccc gtggacaacc gcaagcggtt gcccaccgtc gagcgccttt
     7561 gcccacaacc cggcggccgg ccgcaacaga tcgttttata aatttttttt tttgaaaaag
     7621 aaaaagcccg aaaggcggca acctctcggg cttctggatt tccgatcccc ggaattagat
     7681 cttggcagga tatattgtgg tg
//
