LOCUS       sequence-334470-        7507 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid Double Transcriptional Unit - 
            pG10-90_E-Luc_AtHSP18.2t+pNOS_Red-F_NOSt.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7507)
  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 7507)
  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: pGREAT19
FEATURES             Location/Qualifiers
     source          1..7507
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     196..216
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     promoter        205..251
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     RBS             1738..1746
                     /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      1900..2148
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     promoter        2239..2418
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      4123..4375
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4410..4428)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4617..4641
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4732..5320)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4812..4831)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5491..6282)
                     /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      6926..7361
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    join(7488..7507,1..3)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
ORIGIN
        1 taacggatcc ggtctcagga gatagtttaa actgaaggcg ggaaacgaca atctgatcca
       61 agctcaagct aagcttgcat gcctgcagga tatcgtggat ccaagcttgc cacgtgccgc
      121 cacgtgccgc cacgtgccgc cacgtgcctc tagaggatcc atctccactg acgtaaggga
      181 tgacgcacaa tcccactatc cttcgcaaga cccttcctct atataaggaa gttcatttca
      241 tttggagagg acacgctggg atccaatgga gagagagaag aacgtggtgt acggccccga
      301 gcccaagcac cctctgggca acttcaccgc cggcgagatg ctgtacaacg ctctgcacaa
      361 gcactcccac atcccccagg ccatcctgga cgtgatgggc aacgagtccc tttcctacca
      421 ggagttcttc gacactactg tgaagctggg ccagagcctc cagaactgtg gctacaagat
      481 gaacgatgtc gtgtcgatct gtgcagagaa caacaagaga ttcttcatcc ccatcatctc
      541 cgcctggtac atcggcatgg tggtggcccc tgtgaacgag gactatatcc cagacgagct
      601 gtgtaaagtg accggcatct ccaagccgat cctggtcttt accactagga agatcctgcc
      661 taaggttttg gaggttaaag acagaaccaa ctacataaag aggatcatca tactggactc
      721 tgaagagaac ctgctgggct gcgagagcct gcacaacttc atgtccaggt actccgacaa
      781 caacctccaa acattcaagc ctctgcacta cgaccctgtg gaccaggtag ccgccatcct
      841 gtgctcctcc ggcacaaccg gcctgcctaa aggcgtgatg cagacccaca ggaacatctg
      901 tgtgagactc acacacgcat ctgaccccag agtgggtaca caactcatcc ccggcgtatc
      961 cgtgctggcc tacctgccat tcttccacgc cttcggcttc agtatcaacc tgggctattt
     1021 catggtgggc ctgagagtgg tgatgctccg aaggtttaac caggaggtgt tcctgaaggc
     1081 catccaggac tacgaggtga ggagcgtgat caacgttccc tccacaatcc tgttcctgtc
     1141 caagagccct ctggtggaca agtacgacct atccaccctg gcggagctgt gctgtggagc
     1201 cgctcctctg gcgaaggagg tggccgagat cgccgtgaag aggctgaacc tgccagggat
     1261 acggtgtggc tacggtctaa cagagtctac ctccgccaac atccatactc tgcacaacga
     1321 gttcaagtcc ggctccctgg gcaaggtgac accttacatg gccgccaaga tcatcgacag
     1381 gaacaccggc gaggccctgg gtccaaacca ggtgggcgag ctgtgcatct ggggacctat
     1441 ggtaacaaaa ggctatgtga acaacccaca ggctactaag gaggccatcg acgacgacgg
     1501 ctggctgcac tctggcgact tcggctacta cgacgaggac gagtatttct acatcgtgga
     1561 ccggtacaag gagctgatca aatacaaggg ctatcaggtc gcccctgtgg agctggagga
     1621 gatcctcctt cagcacccag gcatcaggga cgtggccgtc gtgggtatcc ctgacatcga
     1681 ggccggcgag ctgccagccg gcttcgtggt gaagcagccc ggcgcccaac tcaccgctaa
     1741 ggaggtgtac gacttcctgg cccagagggt gtctcactcc aagtacctga ggggcggcgt
     1801 aaggttcgtg gactctatcc ccaggaacgt gacaggcaag attagtcgaa aagagctgag
     1861 ggaggccctg atggagaagg cttctaagct gtaagcttat atgaagatga agatgaaata
     1921 tttggtgtgt caaataaaaa gcttgtgtgc ttaagtttgt gtttttttct tggcttgttg
     1981 tgttatgaat ttgtggcttt ttctaatatt aaatgaatgt aagatctcat tataatgaat
     2041 aaacaaatgt ttctataatc cattgtgaat gttttgttgg atctcttctg cagcatataa
     2101 ctactgtatg tgctatggta tggactatgg aatatgatta aagataagcg ctgtcaggag
     2161 atacatgaga attaagggag tcacggtatg acccccgccg atgacgcggg acaagccgtt
     2221 ttacgtttgg aactgacaga accgcaacgt tgaaggagcc actgagccgc gggtttctgg
     2281 agtttaatga gctaagcaca tacgtcagaa accattattg cgcgttcaaa agtcgcctaa
     2341 ggtcactatc agctagcaaa tatttcttgt caaaaatgct ccactgacgt tccataaatt
     2401 cccctcggta tccaattaga gtctcatatt cactctcaac tcgatcgagg caatggagaa
     2461 catggagaac gacgagaaca ttgtggtggg acccaagccg ttctacccga tcgaggaggg
     2521 ttccgctggc acccagctgc gcaagtacat ggagcgctac gctaagctgg gtgctatcgc
     2581 cttcaccaac gccgtgaccg gtgtggatta ctcctacgcc gagtacctgg agaagtcctg
     2641 ctgcctcgga aaggccctgc agaactacgg tctggtcgtg gatggccgca ttgccctctg
     2701 ctcggagaac tgcgaggagt tcttcattcc cgtgatcgct ggtctgttca tcggagtcgg
     2761 cgttgcccca accaacgaga tctacaccct gcgcgagctg gtccacagcc tgggtatcag
     2821 caagccgacc atcgttttct cgagcaagaa gggcctcgac aaggttatca ccgtgcaaaa
     2881 aaccgtcacc accatcaaga ccatcgttat cctggattcc aaggttgact accgcggtta
     2941 ccagtgcctc gacaccttca tcaagcgtaa caccccaccc ggcttccaag cctcctcctt
     3001 caaaaccgtg gaggttgatc gcaaggagca ggtggccctg atcatgaact cctccggctc
     3061 caccggtctg ccaaagggcg tgcagctgac ccacgagaac accgtcaccc gcttctcgca
     3121 tgcccgtgat cccatctacg gcaaccaggt gtccccaggc accgccgtcc tgaccgtcgt
     3181 gcccctccac cacggcttcg gcatgttcac cactctcggc tacttgatct gcggcttccg
     3241 tgtggtgatg ctgactaagt tcgacgagga gactttcctc aagaccctgc aagactacaa
     3301 gtgcacttac gtgatcctgg tgccgaccct gttcgccatc ctgaacaaga gcgagctgtt
     3361 gaacaaatac gacctgtcga acctggtgga gatcgcttcc ggcggcgccc cactgtcgaa
     3421 ggaagttggc gaggccgttg ctcgccgctt caacctgccc ggcgtccgtc agggttacgg
     3481 cctgaccgag actacctcgg ctatcatcat cacccccgag ggcgacgata agcccggcgc
     3541 ttccggcaag gttgttccac tcttcaaagc caaggtgatc gacctggaca ccaagaagtc
     3601 cctgggtccc aaccgtcgcg gagaagtgtg cgtcaagggc cccatgctga tgaagggcta
     3661 cgtcaacaac ccagaggcca ccaaggagtt gatcgacgag gagggctggc tgcacactgg
     3721 tgacatcggt tactacgacg aagagaagca cttcttcatc gttgaccgcc tgaagtcctt
     3781 gatcaagtac aagggctacc aggttccccc cgccgagctc gagagcgtcc tgctgcagca
     3841 cccctccatt ttcgatgccg gagtggccgg tgtccccgat cccgtggctg gcgagctgcc
     3901 aggtgccgtc gtcgtgctgg agtccggcaa gaacatgacc gagaaggaag tgatggatta
     3961 cgtggcctcg caagtgtcca acgccaagcg cctgcgcggc ggcgtgcgct tcgtggatga
     4021 ggtgcccaag ggcctcaccg gaaagatcga cggtcgcgct atccgcgaga tcctcaagaa
     4081 gcccgtggcc aagatgtgag cttggaatgg atcttcgatc ccgatcgttc aaacatttgg
     4141 caataaagtt tcttaagatt gaatcctgtt gccggtcttg cgacgattat catataattt
     4201 ctgttgaatt acgttaagca tgtaataatt aacatgtaat gcatgacgtt atttatgaga
     4261 tgggttttta tgattagagt cccgcaatta tacatttaat acgcgataga aaacaaaata
     4321 tagcgcgcaa actaggataa attatcgcgc gcggtgtcat ctatgttact agatcgggaa
     4381 ttgccaagct aattcttgaa gatgaaaggg cctcgtgata cgcctatttt tataggttaa
     4441 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     4501 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4561 accctgataa atgcttcaat aatgggaccg actcgcgctg tcatgagacc ggatcctgac
     4621 aggatatatt ggcgggtaaa ctaagtcgct gtatgtgttt gtttgagatc tcatgtgagc
     4681 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4741 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4801 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4861 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4921 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4981 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5041 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5101 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5161 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaag
     5221 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5281 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5341 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5401 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5461 aagtatatat gtgtaacatt ggtctagtga ttatttgccg actaccttgg tgatctcgcc
     5521 tttcacgtag tggacaaatt cttccaactg atctgcgcgc gaggccaagc gatcttcttc
     5581 ttgtccaaga taagcctgtc tagcttcaag tatgacgggc tgatactggg ccggcaggcg
     5641 ctccattgcc cagtcggcag cgacatcctt cggcgcgatt ttgccggtta ctgcgctgta
     5701 ccaaatgcgg gacaacgtaa gcactacatt tcgctcatca ccagcccagt cgggcggcga
     5761 gttccatagc gttaaggttt catttagcgc ctcaaataga tcctgttcag gaaccggatc
     5821 aaagagttcc tccgccgctg gacctaccaa ggcaacgcta tgttctcttg cttttgtcag
     5881 caagatagcc agatcaatgt cgatcgtggc tggctcgaag atacctgcaa gaatgtcatt
     5941 gcgctgccat tctccaaatt gcagttcgcg cttagctgga taacgccacg gaatgatgtc
     6001 gtcgtgcaca acaatggtga cttctacagc gcggagaatc tcgctctctc caggggaagc
     6061 cgaagtttcc aaaaggtcgt tgatcaaagc tcgccgcgtt gtttcatcaa gccttacggt
     6121 caccgtaacc agcaaatcaa tatcactgtg tggcttcagg ccgccatcca ctgcggagcc
     6181 gtacaaatgt acggccagca acgtcggttc gagatggcgc tcgatgacgc caactacctc
     6241 tgatagttga gtcgatactt cggcgatcac cgcttccctc ataatgttta actttgtttt
     6301 agggcgactg ccctgctgcg taacatcgtt gctgctccat aacatcaaac atcgacccac
     6361 ggcgtaacgc gcttgctgct tggatgcccg aggcatagac tgtaccccaa aaaaacagtc
     6421 ataacaagcc atgaaaaccg ccactgcgcc gttaccaccg ctgcgttcgg tcaaggttct
     6481 ggaccagttg cgtgagcgca tacgctactt gcattacagc ttacgaaccg aacaggctta
     6541 tgtccactgg gttcgtgcct tcatccgttt ccacggtgtg cgtcacccgg caaccttggg
     6601 tagcagcgaa gtcgaggcat ttctgtcctg gctggaacac cccttgtatt actgtttatg
     6661 taagcagaca gttttattgt tcatgatgat atatttttat cttgtgcaat gtaacatcag
     6721 agattttgag acacaacgtg gctttgttga ataaatcgaa cttttgctga gttgaaggat
     6781 cagatcacgc atcttcccga caacgcagac cgttccgtgg caaagcaaaa gttcaaaatc
     6841 accaactggt ccacctacaa caaagctctc atcaaccgtg gctccctcac tttctggctg
     6901 gatgatgggg cgattcaggc gatccccatc caacagcccg ccgtcgagcg ggctttttta
     6961 tccccggaag cctgtggata gagggtagtt atccacgtga aaccgctaat gccccgcaaa
     7021 gccttgattc acggggcttt ccggcccgct ccaaaaacta tccacgtgaa atcgctaatc
     7081 agggtacgtg aaatcgctaa tcggagtacg tgaaatcgct aataaggtca cgtgaaatcg
     7141 ctaatcaaaa aggcacgtga gaacgctaat agccctttca gatcaacagc ttgcaaacac
     7201 ccctcgctcc ggcaagtagt tacagcaagt agtatgttca attagctttt caattatgaa
     7261 tatatatatc aattattggt cgcccttggc ttgtggacaa tgcgctacgc gcaccggctc
     7321 cgcccgtgga caaccgcaag cggttgccca ccgtcgagcg cctttgccca caacccggcg
     7381 gccggccgca acagatcgtt ttataaattt ttttttttga aaaagaaaaa gcccgaaagg
     7441 cggcaacctc tcgggcttct ggatttccga tccccggaat tagatcttgg caggatatat
     7501 tgtggtg
//
