LOCUS       sequence-335137-        7797 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid Double Transcriptional Unit - 
            pNOS_E-Luc_NOSt+pNOS_Red-F_NOSt.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7797)
  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 7797)
  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: pGREAT1
FEATURES             Location/Qualifiers
     source          1..7797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        100..279
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     RBS             1786..1794
                     /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      1966..2218
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        2529..2708
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      4413..4665
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    4907..4931
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5022..5610)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5102..5121)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5781..6572)
                     /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      7216..7651
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    join(7778..7797,1..3)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
ORIGIN
        1 taacggatcc ggtctcagga gatacatgag aattaaggga gtcacggtat gacccccgcc
       61 gatgacgcgg gacaagccgt tttacgtttg gaactgacag aaccgcaacg ttgaaggagc
      121 cactgagccg cgggtttctg gagtttaatg agctaagcac atacgtcaga aaccattatt
      181 gcgcgttcaa aagtcgccta aggtcactat cagctagcaa atatttcttg tcaaaaatgc
      241 tccactgacg ttccataaat tcccctcggt atccaattag agtctcatat tcactctcaa
      301 ctcgatcgag gcaatggaga gagagaagaa cgtggtgtac ggccccgagc ccaagcaccc
      361 tctgggcaac ttcaccgccg gcgagatgct gtacaacgct ctgcacaagc actcccacat
      421 cccccaggcc atcctggacg tgatgggcaa cgagtccctt tcctaccagg agttcttcga
      481 cactactgtg aagctgggcc agagcctcca gaactgtggc tacaagatga acgatgtcgt
      541 gtcgatctgt gcagagaaca acaagagatt cttcatcccc atcatctccg cctggtacat
      601 cggcatggtg gtggcccctg tgaacgagga ctatatccca gacgagctgt gtaaagtgac
      661 cggcatctcc aagccgatcc tggtctttac cactaggaag atcctgccta aggttttgga
      721 ggttaaagac agaaccaact acataaagag gatcatcata ctggactctg aagagaacct
      781 gctgggctgc gagagcctgc acaacttcat gtccaggtac tccgacaaca acctccaaac
      841 attcaagcct ctgcactacg accctgtgga ccaggtagcc gccatcctgt gctcctccgg
      901 cacaaccggc ctgcctaaag gcgtgatgca gacccacagg aacatctgtg tgagactcac
      961 acacgcatct gaccccagag tgggtacaca actcatcccc ggcgtatccg tgctggccta
     1021 cctgccattc ttccacgcct tcggcttcag tatcaacctg ggctatttca tggtgggcct
     1081 gagagtggtg atgctccgaa ggtttaacca ggaggtgttc ctgaaggcca tccaggacta
     1141 cgaggtgagg agcgtgatca acgttccctc cacaatcctg ttcctgtcca agagccctct
     1201 ggtggacaag tacgacctat ccaccctggc ggagctgtgc tgtggagccg ctcctctggc
     1261 gaaggaggtg gccgagatcg ccgtgaagag gctgaacctg ccagggatac ggtgtggcta
     1321 cggtctaaca gagtctacct ccgccaacat ccatactctg cacaacgagt tcaagtccgg
     1381 ctccctgggc aaggtgacac cttacatggc cgccaagatc atcgacagga acaccggcga
     1441 ggccctgggt ccaaaccagg tgggcgagct gtgcatctgg ggacctatgg taacaaaagg
     1501 ctatgtgaac aacccacagg ctactaagga ggccatcgac gacgacggct ggctgcactc
     1561 tggcgacttc ggctactacg acgaggacga gtatttctac atcgtggacc ggtacaagga
     1621 gctgatcaaa tacaagggct atcaggtcgc ccctgtggag ctggaggaga tcctccttca
     1681 gcacccaggc atcagggacg tggccgtcgt gggtatccct gacatcgagg ccggcgagct
     1741 gccagccggc ttcgtggtga agcagcccgg cgcccaactc accgctaagg aggtgtacga
     1801 cttcctggcc cagagggtgt ctcactccaa gtacctgagg ggcggcgtaa ggttcgtgga
     1861 ctctatcccc aggaacgtga caggcaagat tagtcgaaaa gagctgaggg aggccctgat
     1921 ggagaaggct tctaagctgt aagcttggaa tggatcttcg atcccgatcg ttcaaacatt
     1981 tggcaataaa gtttcttaag attgaatcct gttgccggtc ttgcgacgat tatcatataa
     2041 tttctgttga attacgttaa gcatgtaata attaacatgt aatgcatgac gttatttatg
     2101 agatgggttt ttatgattag agtcccgcaa ttatacattt aatacgcgat agaaaacaaa
     2161 atatagcgcg caaactagga taaattatcg cgcgcggtgt catctatgtt actagatcgg
     2221 gaattgccaa gctaattctt gaagatgaaa gggcctcgtg atacgcctat ttttataggt
     2281 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     2341 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     2401 ataaccctga taaatgcttc aataatggga ccgactcgcg ctgtcaggag atacatgaga
     2461 attaagggag tcacggtatg acccccgccg atgacgcggg acaagccgtt ttacgtttgg
     2521 aactgacaga accgcaacgt tgaaggagcc actgagccgc gggtttctgg agtttaatga
     2581 gctaagcaca tacgtcagaa accattattg cgcgttcaaa agtcgcctaa ggtcactatc
     2641 agctagcaaa tatttcttgt caaaaatgct ccactgacgt tccataaatt cccctcggta
     2701 tccaattaga gtctcatatt cactctcaac tcgatcgagg caatggagaa catggagaac
     2761 gacgagaaca ttgtggtggg acccaagccg ttctacccga tcgaggaggg ttccgctggc
     2821 acccagctgc gcaagtacat ggagcgctac gctaagctgg gtgctatcgc cttcaccaac
     2881 gccgtgaccg gtgtggatta ctcctacgcc gagtacctgg agaagtcctg ctgcctcgga
     2941 aaggccctgc agaactacgg tctggtcgtg gatggccgca ttgccctctg ctcggagaac
     3001 tgcgaggagt tcttcattcc cgtgatcgct ggtctgttca tcggagtcgg cgttgcccca
     3061 accaacgaga tctacaccct gcgcgagctg gtccacagcc tgggtatcag caagccgacc
     3121 atcgttttct cgagcaagaa gggcctcgac aaggttatca ccgtgcaaaa aaccgtcacc
     3181 accatcaaga ccatcgttat cctggattcc aaggttgact accgcggtta ccagtgcctc
     3241 gacaccttca tcaagcgtaa caccccaccc ggcttccaag cctcctcctt caaaaccgtg
     3301 gaggttgatc gcaaggagca ggtggccctg atcatgaact cctccggctc caccggtctg
     3361 ccaaagggcg tgcagctgac ccacgagaac accgtcaccc gcttctcgca tgcccgtgat
     3421 cccatctacg gcaaccaggt gtccccaggc accgccgtcc tgaccgtcgt gcccctccac
     3481 cacggcttcg gcatgttcac cactctcggc tacttgatct gcggcttccg tgtggtgatg
     3541 ctgactaagt tcgacgagga gactttcctc aagaccctgc aagactacaa gtgcacttac
     3601 gtgatcctgg tgccgaccct gttcgccatc ctgaacaaga gcgagctgtt gaacaaatac
     3661 gacctgtcga acctggtgga gatcgcttcc ggcggcgccc cactgtcgaa ggaagttggc
     3721 gaggccgttg ctcgccgctt caacctgccc ggcgtccgtc agggttacgg cctgaccgag
     3781 actacctcgg ctatcatcat cacccccgag ggcgacgata agcccggcgc ttccggcaag
     3841 gttgttccac tcttcaaagc caaggtgatc gacctggaca ccaagaagtc cctgggtccc
     3901 aaccgtcgcg gagaagtgtg cgtcaagggc cccatgctga tgaagggcta cgtcaacaac
     3961 ccagaggcca ccaaggagtt gatcgacgag gagggctggc tgcacactgg tgacatcggt
     4021 tactacgacg aagagaagca cttcttcatc gttgaccgcc tgaagtcctt gatcaagtac
     4081 aagggctacc aggttccccc cgccgagctc gagagcgtcc tgctgcagca cccctccatt
     4141 ttcgatgccg gagtggccgg tgtccccgat cccgtggctg gcgagctgcc aggtgccgtc
     4201 gtcgtgctgg agtccggcaa gaacatgacc gagaaggaag tgatggatta cgtggcctcg
     4261 caagtgtcca acgccaagcg cctgcgcggc ggcgtgcgct tcgtggatga ggtgcccaag
     4321 ggcctcaccg gaaagatcga cggtcgcgct atccgcgaga tcctcaagaa gcccgtggcc
     4381 aagatgtgag cttggaatgg atcttcgatc ccgatcgttc aaacatttgg caataaagtt
     4441 tcttaagatt gaatcctgtt gccggtcttg cgacgattat catataattt ctgttgaatt
     4501 acgttaagca tgtaataatt aacatgtaat gcatgacgtt atttatgaga tgggttttta
     4561 tgattagagt cccgcaatta tacatttaat acgcgataga aaacaaaata tagcgcgcaa
     4621 actaggataa attatcgcgc gcggtgtcat ctatgttact agatcgggaa ttgccaagct
     4681 aattcttgaa gatgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     4741 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4801 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4861 atgcttcaat aatgggaccg actcgcgctg tcatgagacc ggatcctgac aggatatatt
     4921 ggcgggtaaa ctaagtcgct gtatgtgttt gtttgagatc tcatgtgagc aaaaggccag
     4981 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5041 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5101 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5161 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5221 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5281 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5341 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5401 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5461 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaag aagagttggt
     5521 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5581 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5641 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5701 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5761 gtgtaacatt ggtctagtga ttatttgccg actaccttgg tgatctcgcc tttcacgtag
     5821 tggacaaatt cttccaactg atctgcgcgc gaggccaagc gatcttcttc ttgtccaaga
     5881 taagcctgtc tagcttcaag tatgacgggc tgatactggg ccggcaggcg ctccattgcc
     5941 cagtcggcag cgacatcctt cggcgcgatt ttgccggtta ctgcgctgta ccaaatgcgg
     6001 gacaacgtaa gcactacatt tcgctcatca ccagcccagt cgggcggcga gttccatagc
     6061 gttaaggttt catttagcgc ctcaaataga tcctgttcag gaaccggatc aaagagttcc
     6121 tccgccgctg gacctaccaa ggcaacgcta tgttctcttg cttttgtcag caagatagcc
     6181 agatcaatgt cgatcgtggc tggctcgaag atacctgcaa gaatgtcatt gcgctgccat
     6241 tctccaaatt gcagttcgcg cttagctgga taacgccacg gaatgatgtc gtcgtgcaca
     6301 acaatggtga cttctacagc gcggagaatc tcgctctctc caggggaagc cgaagtttcc
     6361 aaaaggtcgt tgatcaaagc tcgccgcgtt gtttcatcaa gccttacggt caccgtaacc
     6421 agcaaatcaa tatcactgtg tggcttcagg ccgccatcca ctgcggagcc gtacaaatgt
     6481 acggccagca acgtcggttc gagatggcgc tcgatgacgc caactacctc tgatagttga
     6541 gtcgatactt cggcgatcac cgcttccctc ataatgttta actttgtttt agggcgactg
     6601 ccctgctgcg taacatcgtt gctgctccat aacatcaaac atcgacccac ggcgtaacgc
     6661 gcttgctgct tggatgcccg aggcatagac tgtaccccaa aaaaacagtc ataacaagcc
     6721 atgaaaaccg ccactgcgcc gttaccaccg ctgcgttcgg tcaaggttct ggaccagttg
     6781 cgtgagcgca tacgctactt gcattacagc ttacgaaccg aacaggctta tgtccactgg
     6841 gttcgtgcct tcatccgttt ccacggtgtg cgtcacccgg caaccttggg tagcagcgaa
     6901 gtcgaggcat ttctgtcctg gctggaacac cccttgtatt actgtttatg taagcagaca
     6961 gttttattgt tcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag
     7021 acacaacgtg gctttgttga ataaatcgaa cttttgctga gttgaaggat cagatcacgc
     7081 atcttcccga caacgcagac cgttccgtgg caaagcaaaa gttcaaaatc accaactggt
     7141 ccacctacaa caaagctctc atcaaccgtg gctccctcac tttctggctg gatgatgggg
     7201 cgattcaggc gatccccatc caacagcccg ccgtcgagcg ggctttttta tccccggaag
     7261 cctgtggata gagggtagtt atccacgtga aaccgctaat gccccgcaaa gccttgattc
     7321 acggggcttt ccggcccgct ccaaaaacta tccacgtgaa atcgctaatc agggtacgtg
     7381 aaatcgctaa tcggagtacg tgaaatcgct aataaggtca cgtgaaatcg ctaatcaaaa
     7441 aggcacgtga gaacgctaat agccctttca gatcaacagc ttgcaaacac ccctcgctcc
     7501 ggcaagtagt tacagcaagt agtatgttca attagctttt caattatgaa tatatatatc
     7561 aattattggt cgcccttggc ttgtggacaa tgcgctacgc gcaccggctc cgcccgtgga
     7621 caaccgcaag cggttgccca ccgtcgagcg cctttgccca caacccggcg gccggccgca
     7681 acagatcgtt ttataaattt ttttttttga aaaagaaaaa gcccgaaagg cggcaacctc
     7741 tcgggcttct ggatttccga tccccggaat tagatcttgg caggatatat tgtggtg
//
