LOCUS       sequence-334471-        5040 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid Transcriptional Unit - pNOS_Red-F_NOSt Reverse 
            orientation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5040)
  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 5040)
  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: pDGB1a2R+pNOS_Red-F_NOSt
FEATURES             Location/Qualifiers
     source          1..5040
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      147..582
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     primer_bind     complement(862..881)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             873..1688
                     /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             873..1688
                     /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     complement(946..965)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1859..2447
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2348..2367
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2538..2562
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     promoter        2660..2839
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      4544..4796
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4831..4849)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    join(5038..5040,1..20)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
ORIGIN
        1 caccacaata tatcctgcca agatctaatt ccggggatcg gaaatccaga agcccgagag
       61 gttgccgcct ttcgggcttt ttctttttca aaaaaaaaaa tttataaaac gatctgttgc
      121 ggccggccgc cgggttgtgg gcaaaggcgc tcgacggtgg gcaaccgctt gcggttgtcc
      181 acgggcggag ccggtgcgcg tagcgcattg tccacaagcc aagggcgacc aataattgat
      241 atatatattc ataattgaaa agctaattga acatactact tgctgtaact acttgccgga
      301 gcgaggggtg tttgcaagct gttgatctga aagggctatt agcgttctca cgtgcctttt
      361 tgattagcga tttcacgtga ccttattagc gatttcacgt actccgatta gcgatttcac
      421 gtaccctgat tagcgatttc acgtggatag tttttggagc gggccggaaa gccccgtgaa
      481 tcaaggcttt gcggggcatt agcggtttca cgtggataac taccctctat ccacaggctt
      541 ccggggataa aaaagcccgc tcgacggcgg gctgttggat ggggatcgcc tgaatcgccc
      601 catcatccag ccagaaagtg agggagccac ggttgatgag agctttgttg taggtggacc
      661 agttggtgat tttgaacttt tgctttgcca cggaacggtc tgcgttgtcg ggaagatgcg
      721 tgatctgatc cttcaactca gcaaaagttc gatttattca acaaagccac gttgtgtctc
      781 aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt
      841 ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtctt
      901 gctcaaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
      961 gcgataatgt cgggcaatca ggtgcgacaa tctaccgatt gtatgggaag cccgatgcgc
     1021 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
     1081 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
     1141 ctcctgatga tgcatggtta ctcaccactg cgatcccagg gaaaacagca ttccaggtat
     1201 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
     1261 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgactcg
     1321 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
     1381 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac
     1441 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
     1501 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
     1561 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
     1621 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt
     1681 ttttctaatc actagaccaa tgttacacat atatacttta gattgattta aaacttcatt
     1741 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     1801 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     1861 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     1921 cggtggtttg tttgccggat caagagctac caactcttct tccgaaggta actggcttca
     1981 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     2041 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     2101 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     2161 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     2221 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     2281 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     2341 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     2401 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     2461 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgaga tctcaaacaa
     2521 acacatacag cgacttagtt tacccgccaa tatatcctgt cagatatcgt ctcaagcgga
     2581 gatacatgag aattaaggga gtcacggtat gacccccgcc gatgacgcgg gacaagccgt
     2641 tttacgtttg gaactgacag aaccgcaacg ttgaaggagc cactgagccg cgggtttctg
     2701 gagtttaatg agctaagcac atacgtcaga aaccattatt gcgcgttcaa aagtcgccta
     2761 aggtcactat cagctagcaa atatttcttg tcaaaaatgc tccactgacg ttccataaat
     2821 tcccctcggt atccaattag agtctcatat tcactctcaa ctcgatcgag gcaatggaga
     2881 acatggagaa cgacgagaac attgtggtgg gacccaagcc gttctacccg atcgaggagg
     2941 gttccgctgg cacccagctg cgcaagtaca tggagcgcta cgctaagctg ggtgctatcg
     3001 ccttcaccaa cgccgtgacc ggtgtggatt actcctacgc cgagtacctg gagaagtcct
     3061 gctgcctcgg aaaggccctg cagaactacg gtctggtcgt ggatggccgc attgccctct
     3121 gctcggagaa ctgcgaggag ttcttcattc ccgtgatcgc tggtctgttc atcggagtcg
     3181 gcgttgcccc aaccaacgag atctacaccc tgcgcgagct ggtccacagc ctgggtatca
     3241 gcaagccgac catcgttttc tcgagcaaga agggcctcga caaggttatc accgtgcaaa
     3301 aaaccgtcac caccatcaag accatcgtta tcctggattc caaggttgac taccgcggtt
     3361 accagtgcct cgacaccttc atcaagcgta acaccccacc cggcttccaa gcctcctcct
     3421 tcaaaaccgt ggaggttgat cgcaaggagc aggtggccct gatcatgaac tcctccggct
     3481 ccaccggtct gccaaagggc gtgcagctga cccacgagaa caccgtcacc cgcttctcgc
     3541 atgcccgtga tcccatctac ggcaaccagg tgtccccagg caccgccgtc ctgaccgtcg
     3601 tgcccctcca ccacggcttc ggcatgttca ccactctcgg ctacttgatc tgcggcttcc
     3661 gtgtggtgat gctgactaag ttcgacgagg agactttcct caagaccctg caagactaca
     3721 agtgcactta cgtgatcctg gtgccgaccc tgttcgccat cctgaacaag agcgagctgt
     3781 tgaacaaata cgacctgtcg aacctggtgg agatcgcttc cggcggcgcc ccactgtcga
     3841 aggaagttgg cgaggccgtt gctcgccgct tcaacctgcc cggcgtccgt cagggttacg
     3901 gcctgaccga gactacctcg gctatcatca tcacccccga gggcgacgat aagcccggcg
     3961 cttccggcaa ggttgttcca ctcttcaaag ccaaggtgat cgacctggac accaagaagt
     4021 ccctgggtcc caaccgtcgc ggagaagtgt gcgtcaaggg ccccatgctg atgaagggct
     4081 acgtcaacaa cccagaggcc accaaggagt tgatcgacga ggagggctgg ctgcacactg
     4141 gtgacatcgg ttactacgac gaagagaagc acttcttcat cgttgaccgc ctgaagtcct
     4201 tgatcaagta caagggctac caggttcccc ccgccgagct cgagagcgtc ctgctgcagc
     4261 acccctccat tttcgatgcc ggagtggccg gtgtccccga tcccgtggct ggcgagctgc
     4321 caggtgccgt cgtcgtgctg gagtccggca agaacatgac cgagaaggaa gtgatggatt
     4381 acgtggcctc gcaagtgtcc aacgccaagc gcctgcgcgg cggcgtgcgc ttcgtggatg
     4441 aggtgcccaa gggcctcacc ggaaagatcg acggtcgcgc tatccgcgag atcctcaaga
     4501 agcccgtggc caagatgtga gcttggaatg gatcttcgat cccgatcgtt caaacatttg
     4561 gcaataaagt ttcttaagat tgaatcctgt tgccggtctt gcgacgatta tcatataatt
     4621 tctgttgaat tacgttaagc atgtaataat taacatgtaa tgcatgacgt tatttatgag
     4681 atgggttttt atgattagag tcccgcaatt atacatttaa tacgcgatag aaaacaaaat
     4741 atagcgcgca aactaggata aattatcgcg cgcggtgtca tctatgttac tagatcggga
     4801 attgccaagc taattcttga agatgaaagg gcctcgtgat acgcctattt ttataggtta
     4861 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     4921 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4981 aaccctgata aatgcttcaa taatgggacc gactcgcgct tgactgagac gatatcgtta
//
