LOCUS       sequence-334505-        7900 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid Double Transcriptional Unit - 
            pAtUBQ10_E-Luc_AtHSP18.2t+pNOS_Red-F_NOSt.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7900)
  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 7900)
  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: pGREAT18
FEATURES             Location/Qualifiers
     source          1..7900
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             2131..2139
                     /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      2293..2541
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     promoter        2632..2811
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      4516..4768
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4803..4821)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5010..5034
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5125..5713)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5205..5224)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5884..6675)
                     /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      7319..7754
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    join(7881..7900,1..3)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
ORIGIN
        1 taacggatcc ggtctcagga ggtcgacgag tcagtaataa acggcgtcaa agtggttgca
       61 gccggcacac acgagtcgtg tttatcaact caaagcacaa atacttttcc tcaacctaaa
      121 aataaggcaa ttagccaaaa acaactttgc gtgtaaacaa cgctcaatac acgtgtcatt
      181 ttattattag ctattgcttc accgccttag ctttctcgtg acctagtcgt cctcgtcttt
      241 tcttcttctt cttctataaa acaataccca aagagctctt cttcttcaca attcagattt
      301 caatttctca aaatcttaaa aactttctct caattctctc taccgtgatc aaggtaaatt
      361 tctgtgttcc ttattctctc aaaatcttcg attttgtttt cgttcgatcc caatttcgta
      421 tatgttcttt ggtttagatt ctgttaatct tagatcgaag atgattttct gggtttgatc
      481 gttagatatc atcttaattc tcgattaggg tttcatagat atcatccgat ttgttcaaat
      541 aatttgagtt ttgtcgaata attactcttc gatttgtgat ttctatctag atctggtgtt
      601 agtttctagt ttgtgcgatc gaatttgtcg attaatctga gtttttctga ttaacagaat
      661 ggagagagag aagaacgtgg tgtacggccc cgagcccaag caccctctgg gcaacttcac
      721 cgccggcgag atgctgtaca acgctctgca caagcactcc cacatccccc aggccatcct
      781 ggacgtgatg ggcaacgagt ccctttccta ccaggagttc ttcgacacta ctgtgaagct
      841 gggccagagc ctccagaact gtggctacaa gatgaacgat gtcgtgtcga tctgtgcaga
      901 gaacaacaag agattcttca tccccatcat ctccgcctgg tacatcggca tggtggtggc
      961 ccctgtgaac gaggactata tcccagacga gctgtgtaaa gtgaccggca tctccaagcc
     1021 gatcctggtc tttaccacta ggaagatcct gcctaaggtt ttggaggtta aagacagaac
     1081 caactacata aagaggatca tcatactgga ctctgaagag aacctgctgg gctgcgagag
     1141 cctgcacaac ttcatgtcca ggtactccga caacaacctc caaacattca agcctctgca
     1201 ctacgaccct gtggaccagg tagccgccat cctgtgctcc tccggcacaa ccggcctgcc
     1261 taaaggcgtg atgcagaccc acaggaacat ctgtgtgaga ctcacacacg catctgaccc
     1321 cagagtgggt acacaactca tccccggcgt atccgtgctg gcctacctgc cattcttcca
     1381 cgccttcggc ttcagtatca acctgggcta tttcatggtg ggcctgagag tggtgatgct
     1441 ccgaaggttt aaccaggagg tgttcctgaa ggccatccag gactacgagg tgaggagcgt
     1501 gatcaacgtt ccctccacaa tcctgttcct gtccaagagc cctctggtgg acaagtacga
     1561 cctatccacc ctggcggagc tgtgctgtgg agccgctcct ctggcgaagg aggtggccga
     1621 gatcgccgtg aagaggctga acctgccagg gatacggtgt ggctacggtc taacagagtc
     1681 tacctccgcc aacatccata ctctgcacaa cgagttcaag tccggctccc tgggcaaggt
     1741 gacaccttac atggccgcca agatcatcga caggaacacc ggcgaggccc tgggtccaaa
     1801 ccaggtgggc gagctgtgca tctggggacc tatggtaaca aaaggctatg tgaacaaccc
     1861 acaggctact aaggaggcca tcgacgacga cggctggctg cactctggcg acttcggcta
     1921 ctacgacgag gacgagtatt tctacatcgt ggaccggtac aaggagctga tcaaatacaa
     1981 gggctatcag gtcgcccctg tggagctgga ggagatcctc cttcagcacc caggcatcag
     2041 ggacgtggcc gtcgtgggta tccctgacat cgaggccggc gagctgccag ccggcttcgt
     2101 ggtgaagcag cccggcgccc aactcaccgc taaggaggtg tacgacttcc tggcccagag
     2161 ggtgtctcac tccaagtacc tgaggggcgg cgtaaggttc gtggactcta tccccaggaa
     2221 cgtgacaggc aagattagtc gaaaagagct gagggaggcc ctgatggaga aggcttctaa
     2281 gctgtaagct tatatgaaga tgaagatgaa atatttggtg tgtcaaataa aaagcttgtg
     2341 tgcttaagtt tgtgtttttt tcttggcttg ttgtgttatg aatttgtggc tttttctaat
     2401 attaaatgaa tgtaagatct cattataatg aataaacaaa tgtttctata atccattgtg
     2461 aatgttttgt tggatctctt ctgcagcata taactactgt atgtgctatg gtatggacta
     2521 tggaatatga ttaaagataa gcgctgtcag gagatacatg agaattaagg gagtcacggt
     2581 atgacccccg ccgatgacgc gggacaagcc gttttacgtt tggaactgac agaaccgcaa
     2641 cgttgaagga gccactgagc cgcgggtttc tggagtttaa tgagctaagc acatacgtca
     2701 gaaaccatta ttgcgcgttc aaaagtcgcc taaggtcact atcagctagc aaatatttct
     2761 tgtcaaaaat gctccactga cgttccataa attcccctcg gtatccaatt agagtctcat
     2821 attcactctc aactcgatcg aggcaatgga gaacatggag aacgacgaga acattgtggt
     2881 gggacccaag ccgttctacc cgatcgagga gggttccgct ggcacccagc tgcgcaagta
     2941 catggagcgc tacgctaagc tgggtgctat cgccttcacc aacgccgtga ccggtgtgga
     3001 ttactcctac gccgagtacc tggagaagtc ctgctgcctc ggaaaggccc tgcagaacta
     3061 cggtctggtc gtggatggcc gcattgccct ctgctcggag aactgcgagg agttcttcat
     3121 tcccgtgatc gctggtctgt tcatcggagt cggcgttgcc ccaaccaacg agatctacac
     3181 cctgcgcgag ctggtccaca gcctgggtat cagcaagccg accatcgttt tctcgagcaa
     3241 gaagggcctc gacaaggtta tcaccgtgca aaaaaccgtc accaccatca agaccatcgt
     3301 tatcctggat tccaaggttg actaccgcgg ttaccagtgc ctcgacacct tcatcaagcg
     3361 taacacccca cccggcttcc aagcctcctc cttcaaaacc gtggaggttg atcgcaagga
     3421 gcaggtggcc ctgatcatga actcctccgg ctccaccggt ctgccaaagg gcgtgcagct
     3481 gacccacgag aacaccgtca cccgcttctc gcatgcccgt gatcccatct acggcaacca
     3541 ggtgtcccca ggcaccgccg tcctgaccgt cgtgcccctc caccacggct tcggcatgtt
     3601 caccactctc ggctacttga tctgcggctt ccgtgtggtg atgctgacta agttcgacga
     3661 ggagactttc ctcaagaccc tgcaagacta caagtgcact tacgtgatcc tggtgccgac
     3721 cctgttcgcc atcctgaaca agagcgagct gttgaacaaa tacgacctgt cgaacctggt
     3781 ggagatcgct tccggcggcg ccccactgtc gaaggaagtt ggcgaggccg ttgctcgccg
     3841 cttcaacctg cccggcgtcc gtcagggtta cggcctgacc gagactacct cggctatcat
     3901 catcaccccc gagggcgacg ataagcccgg cgcttccggc aaggttgttc cactcttcaa
     3961 agccaaggtg atcgacctgg acaccaagaa gtccctgggt cccaaccgtc gcggagaagt
     4021 gtgcgtcaag ggccccatgc tgatgaaggg ctacgtcaac aacccagagg ccaccaagga
     4081 gttgatcgac gaggagggct ggctgcacac tggtgacatc ggttactacg acgaagagaa
     4141 gcacttcttc atcgttgacc gcctgaagtc cttgatcaag tacaagggct accaggttcc
     4201 ccccgccgag ctcgagagcg tcctgctgca gcacccctcc attttcgatg ccggagtggc
     4261 cggtgtcccc gatcccgtgg ctggcgagct gccaggtgcc gtcgtcgtgc tggagtccgg
     4321 caagaacatg accgagaagg aagtgatgga ttacgtggcc tcgcaagtgt ccaacgccaa
     4381 gcgcctgcgc ggcggcgtgc gcttcgtgga tgaggtgccc aagggcctca ccggaaagat
     4441 cgacggtcgc gctatccgcg agatcctcaa gaagcccgtg gccaagatgt gagcttggaa
     4501 tggatcttcg atcccgatcg ttcaaacatt tggcaataaa gtttcttaag attgaatcct
     4561 gttgccggtc ttgcgacgat tatcatataa tttctgttga attacgttaa gcatgtaata
     4621 attaacatgt aatgcatgac gttatttatg agatgggttt ttatgattag agtcccgcaa
     4681 ttatacattt aatacgcgat agaaaacaaa atatagcgcg caaactagga taaattatcg
     4741 cgcgcggtgt catctatgtt actagatcgg gaattgccaa gctaattctt gaagatgaaa
     4801 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     4861 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     4921 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatggga
     4981 ccgactcgcg ctgtcatgag accggatcct gacaggatat attggcgggt aaactaagtc
     5041 gctgtatgtg tttgtttgag atctcatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5101 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5161 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5221 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5281 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5341 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5401 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5461 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5521 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5581 ctgcgctctg ctgaagccag ttaccttcgg aagaagagtt ggtagctctt gatccggcaa
     5641 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5701 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5761 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5821 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgtgtaac attggtctag
     5881 tgattatttg ccgactacct tggtgatctc gcctttcacg tagtggacaa attcttccaa
     5941 ctgatctgcg cgcgaggcca agcgatcttc ttcttgtcca agataagcct gtctagcttc
     6001 aagtatgacg ggctgatact gggccggcag gcgctccatt gcccagtcgg cagcgacatc
     6061 cttcggcgcg attttgccgg ttactgcgct gtaccaaatg cgggacaacg taagcactac
     6121 atttcgctca tcaccagccc agtcgggcgg cgagttccat agcgttaagg tttcatttag
     6181 cgcctcaaat agatcctgtt caggaaccgg atcaaagagt tcctccgccg ctggacctac
     6241 caaggcaacg ctatgttctc ttgcttttgt cagcaagata gccagatcaa tgtcgatcgt
     6301 ggctggctcg aagatacctg caagaatgtc attgcgctgc cattctccaa attgcagttc
     6361 gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc acaacaatgg tgacttctac
     6421 agcgcggaga atctcgctct ctccagggga agccgaagtt tccaaaaggt cgttgatcaa
     6481 agctcgccgc gttgtttcat caagccttac ggtcaccgta accagcaaat caatatcact
     6541 gtgtggcttc aggccgccat ccactgcgga gccgtacaaa tgtacggcca gcaacgtcgg
     6601 ttcgagatgg cgctcgatga cgccaactac ctctgatagt tgagtcgata cttcggcgat
     6661 caccgcttcc ctcataatgt ttaactttgt tttagggcga ctgccctgct gcgtaacatc
     6721 gttgctgctc cataacatca aacatcgacc cacggcgtaa cgcgcttgct gcttggatgc
     6781 ccgaggcata gactgtaccc caaaaaaaca gtcataacaa gccatgaaaa ccgccactgc
     6841 gccgttacca ccgctgcgtt cggtcaaggt tctggaccag ttgcgtgagc gcatacgcta
     6901 cttgcattac agcttacgaa ccgaacaggc ttatgtccac tgggttcgtg ccttcatccg
     6961 tttccacggt gtgcgtcacc cggcaacctt gggtagcagc gaagtcgagg catttctgtc
     7021 ctggctggaa caccccttgt attactgttt atgtaagcag acagttttat tgttcatgat
     7081 gatatatttt tatcttgtgc aatgtaacat cagagatttt gagacacaac gtggctttgt
     7141 tgaataaatc gaacttttgc tgagttgaag gatcagatca cgcatcttcc cgacaacgca
     7201 gaccgttccg tggcaaagca aaagttcaaa atcaccaact ggtccaccta caacaaagct
     7261 ctcatcaacc gtggctccct cactttctgg ctggatgatg gggcgattca ggcgatcccc
     7321 atccaacagc ccgccgtcga gcgggctttt ttatccccgg aagcctgtgg atagagggta
     7381 gttatccacg tgaaaccgct aatgccccgc aaagccttga ttcacggggc tttccggccc
     7441 gctccaaaaa ctatccacgt gaaatcgcta atcagggtac gtgaaatcgc taatcggagt
     7501 acgtgaaatc gctaataagg tcacgtgaaa tcgctaatca aaaaggcacg tgagaacgct
     7561 aatagccctt tcagatcaac agcttgcaaa cacccctcgc tccggcaagt agttacagca
     7621 agtagtatgt tcaattagct tttcaattat gaatatatat atcaattatt ggtcgccctt
     7681 ggcttgtgga caatgcgcta cgcgcaccgg ctccgcccgt ggacaaccgc aagcggttgc
     7741 ccaccgtcga gcgcctttgc ccacaacccg gcggccggcc gcaacagatc gttttataaa
     7801 tttttttttt tgaaaaagaa aaagcccgaa aggcggcaac ctctcgggct tctggatttc
     7861 cgatccccgg aattagatct tggcaggata tattgtggtg
//
