LOCUS       sequence-344598-        7944 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid Double Transcriptional Unit - 
            p35SoTMV_E-Luc_NOSt+pNOS_Red-F_NOSt.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7944)
  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 7944)
  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: pGREAT2
FEATURES             Location/Qualifiers
     source          1..7944
                     /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      2113..2365
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        2676..2855
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      4560..4812
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    5054..5078
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5169..5757)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5249..5268)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5928..6719)
                     /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      7363..7798
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    join(7925..7944,1..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 cttggaatgg
     2101 atcttcgatc ccgatcgttc aaacatttgg caataaagtt tcttaagatt gaatcctgtt
     2161 gccggtcttg cgacgattat catataattt ctgttgaatt acgttaagca tgtaataatt
     2221 aacatgtaat gcatgacgtt atttatgaga tgggttttta tgattagagt cccgcaatta
     2281 tacatttaat acgcgataga aaacaaaata tagcgcgcaa actaggataa attatcgcgc
     2341 gcggtgtcat ctatgttact agatcgggaa ttgccaagct aattcttgaa gatgaaaggg
     2401 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     2461 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     2521 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatgggaccg
     2581 actcgcgctg tcaggagata catgagaatt aagggagtca cggtatgacc cccgccgatg
     2641 acgcgggaca agccgtttta cgtttggaac tgacagaacc gcaacgttga aggagccact
     2701 gagccgcggg tttctggagt ttaatgagct aagcacatac gtcagaaacc attattgcgc
     2761 gttcaaaagt cgcctaaggt cactatcagc tagcaaatat ttcttgtcaa aaatgctcca
     2821 ctgacgttcc ataaattccc ctcggtatcc aattagagtc tcatattcac tctcaactcg
     2881 atcgaggcaa tggagaacat ggagaacgac gagaacattg tggtgggacc caagccgttc
     2941 tacccgatcg aggagggttc cgctggcacc cagctgcgca agtacatgga gcgctacgct
     3001 aagctgggtg ctatcgcctt caccaacgcc gtgaccggtg tggattactc ctacgccgag
     3061 tacctggaga agtcctgctg cctcggaaag gccctgcaga actacggtct ggtcgtggat
     3121 ggccgcattg ccctctgctc ggagaactgc gaggagttct tcattcccgt gatcgctggt
     3181 ctgttcatcg gagtcggcgt tgccccaacc aacgagatct acaccctgcg cgagctggtc
     3241 cacagcctgg gtatcagcaa gccgaccatc gttttctcga gcaagaaggg cctcgacaag
     3301 gttatcaccg tgcaaaaaac cgtcaccacc atcaagacca tcgttatcct ggattccaag
     3361 gttgactacc gcggttacca gtgcctcgac accttcatca agcgtaacac cccacccggc
     3421 ttccaagcct cctccttcaa aaccgtggag gttgatcgca aggagcaggt ggccctgatc
     3481 atgaactcct ccggctccac cggtctgcca aagggcgtgc agctgaccca cgagaacacc
     3541 gtcacccgct tctcgcatgc ccgtgatccc atctacggca accaggtgtc cccaggcacc
     3601 gccgtcctga ccgtcgtgcc cctccaccac ggcttcggca tgttcaccac tctcggctac
     3661 ttgatctgcg gcttccgtgt ggtgatgctg actaagttcg acgaggagac tttcctcaag
     3721 accctgcaag actacaagtg cacttacgtg atcctggtgc cgaccctgtt cgccatcctg
     3781 aacaagagcg agctgttgaa caaatacgac ctgtcgaacc tggtggagat cgcttccggc
     3841 ggcgccccac tgtcgaagga agttggcgag gccgttgctc gccgcttcaa cctgcccggc
     3901 gtccgtcagg gttacggcct gaccgagact acctcggcta tcatcatcac ccccgagggc
     3961 gacgataagc ccggcgcttc cggcaaggtt gttccactct tcaaagccaa ggtgatcgac
     4021 ctggacacca agaagtccct gggtcccaac cgtcgcggag aagtgtgcgt caagggcccc
     4081 atgctgatga agggctacgt caacaaccca gaggccacca aggagttgat cgacgaggag
     4141 ggctggctgc acactggtga catcggttac tacgacgaag agaagcactt cttcatcgtt
     4201 gaccgcctga agtccttgat caagtacaag ggctaccagg ttccccccgc cgagctcgag
     4261 agcgtcctgc tgcagcaccc ctccattttc gatgccggag tggccggtgt ccccgatccc
     4321 gtggctggcg agctgccagg tgccgtcgtc gtgctggagt ccggcaagaa catgaccgag
     4381 aaggaagtga tggattacgt ggcctcgcaa gtgtccaacg ccaagcgcct gcgcggcggc
     4441 gtgcgcttcg tggatgaggt gcccaagggc ctcaccggaa agatcgacgg tcgcgctatc
     4501 cgcgagatcc tcaagaagcc cgtggccaag atgtgagctt ggaatggatc ttcgatcccg
     4561 atcgttcaaa catttggcaa taaagtttct taagattgaa tcctgttgcc ggtcttgcga
     4621 cgattatcat ataatttctg ttgaattacg ttaagcatgt aataattaac atgtaatgca
     4681 tgacgttatt tatgagatgg gtttttatga ttagagtccc gcaattatac atttaatacg
     4741 cgatagaaaa caaaatatag cgcgcaaact aggataaatt atcgcgcgcg gtgtcatcta
     4801 tgttactaga tcgggaattg ccaagctaat tcttgaagat gaaagggcct cgtgatacgc
     4861 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     4921 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4981 ccgctcatga gacaataacc ctgataaatg cttcaataat gggaccgact cgcgctgtca
     5041 tgagaccgga tcctgacagg atatattggc gggtaaacta agtcgctgta tgtgtttgtt
     5101 tgagatctca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5161 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5221 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5281 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5341 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5401 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5461 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5521 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5581 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     5641 ccagttacct tcggaagaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5701 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5761 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5821 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5881 agttttaaat caatctaaag tatatatgtg taacattggt ctagtgatta tttgccgact
     5941 accttggtga tctcgccttt cacgtagtgg acaaattctt ccaactgatc tgcgcgcgag
     6001 gccaagcgat cttcttcttg tccaagataa gcctgtctag cttcaagtat gacgggctga
     6061 tactgggccg gcaggcgctc cattgcccag tcggcagcga catccttcgg cgcgattttg
     6121 ccggttactg cgctgtacca aatgcgggac aacgtaagca ctacatttcg ctcatcacca
     6181 gcccagtcgg gcggcgagtt ccatagcgtt aaggtttcat ttagcgcctc aaatagatcc
     6241 tgttcaggaa ccggatcaaa gagttcctcc gccgctggac ctaccaaggc aacgctatgt
     6301 tctcttgctt ttgtcagcaa gatagccaga tcaatgtcga tcgtggctgg ctcgaagata
     6361 cctgcaagaa tgtcattgcg ctgccattct ccaaattgca gttcgcgctt agctggataa
     6421 cgccacggaa tgatgtcgtc gtgcacaaca atggtgactt ctacagcgcg gagaatctcg
     6481 ctctctccag gggaagccga agtttccaaa aggtcgttga tcaaagctcg ccgcgttgtt
     6541 tcatcaagcc ttacggtcac cgtaaccagc aaatcaatat cactgtgtgg cttcaggccg
     6601 ccatccactg cggagccgta caaatgtacg gccagcaacg tcggttcgag atggcgctcg
     6661 atgacgccaa ctacctctga tagttgagtc gatacttcgg cgatcaccgc ttccctcata
     6721 atgtttaact ttgttttagg gcgactgccc tgctgcgtaa catcgttgct gctccataac
     6781 atcaaacatc gacccacggc gtaacgcgct tgctgcttgg atgcccgagg catagactgt
     6841 accccaaaaa aacagtcata acaagccatg aaaaccgcca ctgcgccgtt accaccgctg
     6901 cgttcggtca aggttctgga ccagttgcgt gagcgcatac gctacttgca ttacagctta
     6961 cgaaccgaac aggcttatgt ccactgggtt cgtgccttca tccgtttcca cggtgtgcgt
     7021 cacccggcaa ccttgggtag cagcgaagtc gaggcatttc tgtcctggct ggaacacccc
     7081 ttgtattact gtttatgtaa gcagacagtt ttattgttca tgatgatata tttttatctt
     7141 gtgcaatgta acatcagaga ttttgagaca caacgtggct ttgttgaata aatcgaactt
     7201 ttgctgagtt gaaggatcag atcacgcatc ttcccgacaa cgcagaccgt tccgtggcaa
     7261 agcaaaagtt caaaatcacc aactggtcca cctacaacaa agctctcatc aaccgtggct
     7321 ccctcacttt ctggctggat gatggggcga ttcaggcgat ccccatccaa cagcccgccg
     7381 tcgagcgggc ttttttatcc ccggaagcct gtggatagag ggtagttatc cacgtgaaac
     7441 cgctaatgcc ccgcaaagcc ttgattcacg gggctttccg gcccgctcca aaaactatcc
     7501 acgtgaaatc gctaatcagg gtacgtgaaa tcgctaatcg gagtacgtga aatcgctaat
     7561 aaggtcacgt gaaatcgcta atcaaaaagg cacgtgagaa cgctaatagc cctttcagat
     7621 caacagcttg caaacacccc tcgctccggc aagtagttac agcaagtagt atgttcaatt
     7681 agcttttcaa ttatgaatat atatatcaat tattggtcgc ccttggcttg tggacaatgc
     7741 gctacgcgca ccggctccgc ccgtggacaa ccgcaagcgg ttgcccaccg tcgagcgcct
     7801 ttgcccacaa cccggcggcc ggccgcaaca gatcgtttta taaatttttt tttttgaaaa
     7861 agaaaaagcc cgaaaggcgg caacctctcg ggcttctgga tttccgatcc ccggaattag
     7921 atcttggcag gatatattgt ggtg
//
