LOCUS       sequence-334549-        7678 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid Double Transcriptional Unit - 
            p35SoTMV_E-Luc_35St+pNOS_Red-F_NOSt.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7678)
  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 7678)
  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: pGREAT22
FEATURES             Location/Qualifiers
     source          1..7678
                     /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)"
     promoter        2410..2589
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      4294..4546
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4581..4599)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4788..4812
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4903..5491)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4983..5002)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5662..6453)
                     /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      7097..7532
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    join(7659..7678,1..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 cttcggccat
     2101 gctagagtcc gcaaaaatca ccagtctctc tctacaaatc tatctctctc tatttttctc
     2161 cagaataatg tgtgagtagt tcccagataa gggaattagg gttcttatag ggtttcgctc
     2221 atgtgttgag catataagaa acccttagta tgtatttgta tttgtaaaat acttctatca
     2281 ataaaatttc taattcctaa aaccaaaatc cagtgacctc gctgtcagga gatacatgag
     2341 aattaaggga gtcacggtat gacccccgcc gatgacgcgg gacaagccgt tttacgtttg
     2401 gaactgacag aaccgcaacg ttgaaggagc cactgagccg cgggtttctg gagtttaatg
     2461 agctaagcac atacgtcaga aaccattatt gcgcgttcaa aagtcgccta aggtcactat
     2521 cagctagcaa atatttcttg tcaaaaatgc tccactgacg ttccataaat tcccctcggt
     2581 atccaattag agtctcatat tcactctcaa ctcgatcgag gcaatggaga acatggagaa
     2641 cgacgagaac attgtggtgg gacccaagcc gttctacccg atcgaggagg gttccgctgg
     2701 cacccagctg cgcaagtaca tggagcgcta cgctaagctg ggtgctatcg ccttcaccaa
     2761 cgccgtgacc ggtgtggatt actcctacgc cgagtacctg gagaagtcct gctgcctcgg
     2821 aaaggccctg cagaactacg gtctggtcgt ggatggccgc attgccctct gctcggagaa
     2881 ctgcgaggag ttcttcattc ccgtgatcgc tggtctgttc atcggagtcg gcgttgcccc
     2941 aaccaacgag atctacaccc tgcgcgagct ggtccacagc ctgggtatca gcaagccgac
     3001 catcgttttc tcgagcaaga agggcctcga caaggttatc accgtgcaaa aaaccgtcac
     3061 caccatcaag accatcgtta tcctggattc caaggttgac taccgcggtt accagtgcct
     3121 cgacaccttc atcaagcgta acaccccacc cggcttccaa gcctcctcct tcaaaaccgt
     3181 ggaggttgat cgcaaggagc aggtggccct gatcatgaac tcctccggct ccaccggtct
     3241 gccaaagggc gtgcagctga cccacgagaa caccgtcacc cgcttctcgc atgcccgtga
     3301 tcccatctac ggcaaccagg tgtccccagg caccgccgtc ctgaccgtcg tgcccctcca
     3361 ccacggcttc ggcatgttca ccactctcgg ctacttgatc tgcggcttcc gtgtggtgat
     3421 gctgactaag ttcgacgagg agactttcct caagaccctg caagactaca agtgcactta
     3481 cgtgatcctg gtgccgaccc tgttcgccat cctgaacaag agcgagctgt tgaacaaata
     3541 cgacctgtcg aacctggtgg agatcgcttc cggcggcgcc ccactgtcga aggaagttgg
     3601 cgaggccgtt gctcgccgct tcaacctgcc cggcgtccgt cagggttacg gcctgaccga
     3661 gactacctcg gctatcatca tcacccccga gggcgacgat aagcccggcg cttccggcaa
     3721 ggttgttcca ctcttcaaag ccaaggtgat cgacctggac accaagaagt ccctgggtcc
     3781 caaccgtcgc ggagaagtgt gcgtcaaggg ccccatgctg atgaagggct acgtcaacaa
     3841 cccagaggcc accaaggagt tgatcgacga ggagggctgg ctgcacactg gtgacatcgg
     3901 ttactacgac gaagagaagc acttcttcat cgttgaccgc ctgaagtcct tgatcaagta
     3961 caagggctac caggttcccc ccgccgagct cgagagcgtc ctgctgcagc acccctccat
     4021 tttcgatgcc ggagtggccg gtgtccccga tcccgtggct ggcgagctgc caggtgccgt
     4081 cgtcgtgctg gagtccggca agaacatgac cgagaaggaa gtgatggatt acgtggcctc
     4141 gcaagtgtcc aacgccaagc gcctgcgcgg cggcgtgcgc ttcgtggatg aggtgcccaa
     4201 gggcctcacc ggaaagatcg acggtcgcgc tatccgcgag atcctcaaga agcccgtggc
     4261 caagatgtga gcttggaatg gatcttcgat cccgatcgtt caaacatttg gcaataaagt
     4321 ttcttaagat tgaatcctgt tgccggtctt gcgacgatta tcatataatt tctgttgaat
     4381 tacgttaagc atgtaataat taacatgtaa tgcatgacgt tatttatgag atgggttttt
     4441 atgattagag tcccgcaatt atacatttaa tacgcgatag aaaacaaaat atagcgcgca
     4501 aactaggata aattatcgcg cgcggtgtca tctatgttac tagatcggga attgccaagc
     4561 taattcttga agatgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     4621 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     4681 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4741 aatgcttcaa taatgggacc gactcgcgct gtcatgagac cggatcctga caggatatat
     4801 tggcgggtaa actaagtcgc tgtatgtgtt tgtttgagat ctcatgtgag caaaaggcca
     4861 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4921 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4981 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5041 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5101 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5161 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5221 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5281 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5341 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa gaagagttgg
     5401 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5461 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5521 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5581 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5641 tgtgtaacat tggtctagtg attatttgcc gactaccttg gtgatctcgc ctttcacgta
     5701 gtggacaaat tcttccaact gatctgcgcg cgaggccaag cgatcttctt cttgtccaag
     5761 ataagcctgt ctagcttcaa gtatgacggg ctgatactgg gccggcaggc gctccattgc
     5821 ccagtcggca gcgacatcct tcggcgcgat tttgccggtt actgcgctgt accaaatgcg
     5881 ggacaacgta agcactacat ttcgctcatc accagcccag tcgggcggcg agttccatag
     5941 cgttaaggtt tcatttagcg cctcaaatag atcctgttca ggaaccggat caaagagttc
     6001 ctccgccgct ggacctacca aggcaacgct atgttctctt gcttttgtca gcaagatagc
     6061 cagatcaatg tcgatcgtgg ctggctcgaa gatacctgca agaatgtcat tgcgctgcca
     6121 ttctccaaat tgcagttcgc gcttagctgg ataacgccac ggaatgatgt cgtcgtgcac
     6181 aacaatggtg acttctacag cgcggagaat ctcgctctct ccaggggaag ccgaagtttc
     6241 caaaaggtcg ttgatcaaag ctcgccgcgt tgtttcatca agccttacgg tcaccgtaac
     6301 cagcaaatca atatcactgt gtggcttcag gccgccatcc actgcggagc cgtacaaatg
     6361 tacggccagc aacgtcggtt cgagatggcg ctcgatgacg ccaactacct ctgatagttg
     6421 agtcgatact tcggcgatca ccgcttccct cataatgttt aactttgttt tagggcgact
     6481 gccctgctgc gtaacatcgt tgctgctcca taacatcaaa catcgaccca cggcgtaacg
     6541 cgcttgctgc ttggatgccc gaggcataga ctgtacccca aaaaaacagt cataacaagc
     6601 catgaaaacc gccactgcgc cgttaccacc gctgcgttcg gtcaaggttc tggaccagtt
     6661 gcgtgagcgc atacgctact tgcattacag cttacgaacc gaacaggctt atgtccactg
     6721 ggttcgtgcc ttcatccgtt tccacggtgt gcgtcacccg gcaaccttgg gtagcagcga
     6781 agtcgaggca tttctgtcct ggctggaaca ccccttgtat tactgtttat gtaagcagac
     6841 agttttattg ttcatgatga tatattttta tcttgtgcaa tgtaacatca gagattttga
     6901 gacacaacgt ggctttgttg aataaatcga acttttgctg agttgaagga tcagatcacg
     6961 catcttcccg acaacgcaga ccgttccgtg gcaaagcaaa agttcaaaat caccaactgg
     7021 tccacctaca acaaagctct catcaaccgt ggctccctca ctttctggct ggatgatggg
     7081 gcgattcagg cgatccccat ccaacagccc gccgtcgagc gggctttttt atccccggaa
     7141 gcctgtggat agagggtagt tatccacgtg aaaccgctaa tgccccgcaa agccttgatt
     7201 cacggggctt tccggcccgc tccaaaaact atccacgtga aatcgctaat cagggtacgt
     7261 gaaatcgcta atcggagtac gtgaaatcgc taataaggtc acgtgaaatc gctaatcaaa
     7321 aaggcacgtg agaacgctaa tagccctttc agatcaacag cttgcaaaca cccctcgctc
     7381 cggcaagtag ttacagcaag tagtatgttc aattagcttt tcaattatga atatatatat
     7441 caattattgg tcgcccttgg cttgtggaca atgcgctacg cgcaccggct ccgcccgtgg
     7501 acaaccgcaa gcggttgccc accgtcgagc gcctttgccc acaacccggc ggccggccgc
     7561 aacagatcgt tttataaatt tttttttttg aaaaagaaaa agcccgaaag gcggcaacct
     7621 ctcgggcttc tggatttccg atccccggaa ttagatcttg gcaggatata ttgtggtg
//
