LOCUS       sequence-334421-        5038 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid Transcriptional Unit - pNOS_Red-F_NOSt Forward 
            orientation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5038)
  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 5038)
  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: pDGB1a2+pNOS_Red-F_NOSt
FEATURES             Location/Qualifiers
     source          1..5038
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        104..283
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      1988..2240
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(2275..2293)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    2477..2501
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(2592..3180)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2672..2691)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3351..4166)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3351..4166)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4074..4093
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4158..4177
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4457..4892
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    join(5019..5038,1..3)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
ORIGIN
        1 taacaagctt cgtctcagtc aggagataca tgagaattaa gggagtcacg gtatgacccc
       61 cgccgatgac gcgggacaag ccgttttacg tttggaactg acagaaccgc aacgttgaag
      121 gagccactga gccgcgggtt tctggagttt aatgagctaa gcacatacgt cagaaaccat
      181 tattgcgcgt tcaaaagtcg cctaaggtca ctatcagcta gcaaatattt cttgtcaaaa
      241 atgctccact gacgttccat aaattcccct cggtatccaa ttagagtctc atattcactc
      301 tcaactcgat cgaggcaatg gagaacatgg agaacgacga gaacattgtg gtgggaccca
      361 agccgttcta cccgatcgag gagggttccg ctggcaccca gctgcgcaag tacatggagc
      421 gctacgctaa gctgggtgct atcgccttca ccaacgccgt gaccggtgtg gattactcct
      481 acgccgagta cctggagaag tcctgctgcc tcggaaaggc cctgcagaac tacggtctgg
      541 tcgtggatgg ccgcattgcc ctctgctcgg agaactgcga ggagttcttc attcccgtga
      601 tcgctggtct gttcatcgga gtcggcgttg ccccaaccaa cgagatctac accctgcgcg
      661 agctggtcca cagcctgggt atcagcaagc cgaccatcgt tttctcgagc aagaagggcc
      721 tcgacaaggt tatcaccgtg caaaaaaccg tcaccaccat caagaccatc gttatcctgg
      781 attccaaggt tgactaccgc ggttaccagt gcctcgacac cttcatcaag cgtaacaccc
      841 cacccggctt ccaagcctcc tccttcaaaa ccgtggaggt tgatcgcaag gagcaggtgg
      901 ccctgatcat gaactcctcc ggctccaccg gtctgccaaa gggcgtgcag ctgacccacg
      961 agaacaccgt cacccgcttc tcgcatgccc gtgatcccat ctacggcaac caggtgtccc
     1021 caggcaccgc cgtcctgacc gtcgtgcccc tccaccacgg cttcggcatg ttcaccactc
     1081 tcggctactt gatctgcggc ttccgtgtgg tgatgctgac taagttcgac gaggagactt
     1141 tcctcaagac cctgcaagac tacaagtgca cttacgtgat cctggtgccg accctgttcg
     1201 ccatcctgaa caagagcgag ctgttgaaca aatacgacct gtcgaacctg gtggagatcg
     1261 cttccggcgg cgccccactg tcgaaggaag ttggcgaggc cgttgctcgc cgcttcaacc
     1321 tgcccggcgt ccgtcagggt tacggcctga ccgagactac ctcggctatc atcatcaccc
     1381 ccgagggcga cgataagccc ggcgcttccg gcaaggttgt tccactcttc aaagccaagg
     1441 tgatcgacct ggacaccaag aagtccctgg gtcccaaccg tcgcggagaa gtgtgcgtca
     1501 agggccccat gctgatgaag ggctacgtca acaacccaga ggccaccaag gagttgatcg
     1561 acgaggaggg ctggctgcac actggtgaca tcggttacta cgacgaagag aagcacttct
     1621 tcatcgttga ccgcctgaag tccttgatca agtacaaggg ctaccaggtt ccccccgccg
     1681 agctcgagag cgtcctgctg cagcacccct ccattttcga tgccggagtg gccggtgtcc
     1741 ccgatcccgt ggctggcgag ctgccaggtg ccgtcgtcgt gctggagtcc ggcaagaaca
     1801 tgaccgagaa ggaagtgatg gattacgtgg cctcgcaagt gtccaacgcc aagcgcctgc
     1861 gcggcggcgt gcgcttcgtg gatgaggtgc ccaagggcct caccggaaag atcgacggtc
     1921 gcgctatccg cgagatcctc aagaagcccg tggccaagat gtgagcttgg aatggatctt
     1981 cgatcccgat cgttcaaaca tttggcaata aagtttctta agattgaatc ctgttgccgg
     2041 tcttgcgacg attatcatat aatttctgtt gaattacgtt aagcatgtaa taattaacat
     2101 gtaatgcatg acgttattta tgagatgggt ttttatgatt agagtcccgc aattatacat
     2161 ttaatacgcg atagaaaaca aaatatagcg cgcaaactag gataaattat cgcgcgcggt
     2221 gtcatctatg ttactagatc gggaattgcc aagctaattc ttgaagatga aagggcctcg
     2281 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     2341 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     2401 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatgg gaccgactcg
     2461 cgcttgagac gaagcttgac aggatatatt ggcgggtaaa ctaagtcgct gtatgtgttt
     2521 gtttgagatc tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2581 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2641 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2701 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2761 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     2821 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     2881 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     2941 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3001 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3061 aagccagtta ccttcggaag aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3121 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3181 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3241 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3301 tgaagtttta aatcaatcta aagtatatat gtgtaacatt ggtctagtga ttagaaaaac
     3361 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
     3421 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
     3481 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
     3541 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
     3601 gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc
     3661 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg
     3721 agtcgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
     3781 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
     3841 acctggaatg ctgttttccc tgggatcgca gtggtgagta accatgcatc atcaggagta
     3901 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
     3961 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
     4021 gcatcgggct tcccatacaa tcggtagatt gtcgcacctg attgcccgac attatcgcga
     4081 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg ccttgagcaa
     4141 gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac
     4201 agttttattg ttcatgatga tatattttta tcttgtgcaa tgtaacatca gagattttga
     4261 gacacaacgt ggctttgttg aataaatcga acttttgctg agttgaagga tcagatcacg
     4321 catcttcccg acaacgcaga ccgttccgtg gcaaagcaaa agttcaaaat caccaactgg
     4381 tccacctaca acaaagctct catcaaccgt ggctccctca ctttctggct ggatgatggg
     4441 gcgattcagg cgatccccat ccaacagccc gccgtcgagc gggctttttt atccccggaa
     4501 gcctgtggat agagggtagt tatccacgtg aaaccgctaa tgccccgcaa agccttgatt
     4561 cacggggctt tccggcccgc tccaaaaact atccacgtga aatcgctaat cagggtacgt
     4621 gaaatcgcta atcggagtac gtgaaatcgc taataaggtc acgtgaaatc gctaatcaaa
     4681 aaggcacgtg agaacgctaa tagccctttc agatcaacag cttgcaaaca cccctcgctc
     4741 cggcaagtag ttacagcaag tagtatgttc aattagcttt tcaattatga atatatatat
     4801 caattattgg tcgcccttgg cttgtggaca atgcgctacg cgcaccggct ccgcccgtgg
     4861 acaaccgcaa gcggttgccc accgtcgagc gcctttgccc acaacccggc ggccggccgc
     4921 aacagatcgt tttataaatt tttttttttg aaaaagaaaa agcccgaaag gcggcaacct
     4981 ctcgggcttc tggatttccg atccccggaa ttagatcttg gcaggatata ttgtggtg
//
