LOCUS       sequence-367789-        8825 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transcriptional unit for copper-inducible expression of nanoluc 
            luciferase driven by a minimal synthetic promoter with binding sites
            for CUP2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8825)
  AUTHORS   Kallam K, Moreno-Gimenez E, Mateos-Fernandez R, Tansley C, Gianoglio
            S, Orzaez D, Patron N
  TITLE     Tunable control of insect pheromone biosynthesis in Nicotiana 
            benthamiana.
  JOURNAL   Plant Biotechnol J. 2023 Apr 9. doi: 10.1111/pbi.14048.
  PUBMED    37032497
REFERENCE   2  (bases 1 to 8825)
  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: "Plant 
            Biotechnol J. 2023 Apr 9. doi: 10.1111/pbi.14048."
COMMENT     Sequence Label: pEPCT-alpha-KN0013
FEATURES             Location/Qualifiers
     source          1..8825
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    22..46
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             1347..1976
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             2405..3478
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      3544..3738
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(3974..3996)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4082..4222
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4137..4156
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4237..4254)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4408..4996)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4488..4507)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5084..5899)
                     /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(5084..5899)
                     /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     5807..5826
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    7670..7694
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        7900..7946
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     misc_feature    7962..8017
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     CDS             8019..8531
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             8592..8615
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    join(8653..8825,1..2)
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
ORIGIN
        1 tcgctgtcat gagacgaatt ctgacaggat atattggcgg gtaaacctaa gagaaaagag
       61 cgtttattag aataatcgga tatttaaaag ggcgtgaaaa ggtttatccg ttcgtccatt
      121 tgtatgtgca tgccaaccac agggttcccc tcgggatcaa agtactttga tccaacccct
      181 ccgctgctat agtgcagtcg gcttctgacg ttcagtgcag ccgtcatctg aaaacgacat
      241 gtcgcacaag tcctaagtta cgcgacaggc tgccgccctg cccttttcct ggcgttttct
      301 tgtcgcgtgt tttagtcgca taaagtagaa tacttgcgac tagaaccgga gacattacgc
      361 catgaacaag agcgccgccg ctggcctgct gggctatgcc cgcgtcagca ccgacgacca
      421 ggacttgacc aaccaacggg ccgaactgca cgcggccggc tgcaccaagc tgttttccga
      481 gaagatcacc ggcaccaggc gcgaccgccc ggagctggcc aggatgcttg accacctacg
      541 ccctggcgac gttgtgacag tgaccaggct agaccgcctg gcccgcagca cccgcgacct
      601 actggacatt gccgagcgca tccaggaggc cggcgcgggc ctgcgtagcc tggcagagcc
      661 gtgggccgac accaccacgc cggccggccg catggtgttg accgtgttcg ccggcattgc
      721 cgagttcgag cgttccctaa tcatcgaccg cacccggagc gggcgcgagg ccgccaaggc
      781 ccgaggcgtg aagtttggcc cccgccctac cctcaccccg gcacagatcg cgcacgcccg
      841 cgagctgatc gaccaggaag gccgcaccgt gaaagaggcg gctgcactgc ttggcgtgca
      901 tcgctcgacc ctgtaccgcg cacttgagcg cagcgaggaa gtgacgccca ccgaggccag
      961 gcggcgcggt gccttccgtg aggacgcatt gaccgaggcc gacgccctgg cggccgccga
     1021 gaatgaacgc caagaggaac aagcatgaaa ccgcaccagg acggccagga cgaaccgttt
     1081 ttcattaccg aagagatcga ggcggagatg atcgcggccg ggtacgtgtt cgagccgccc
     1141 gcgcacctct caaccgtgcg gctgcatgaa atcctggccg gtttgtctga tgccaagctg
     1201 gcggcctggc cggccagctt ggccgctgaa gaaaccgagc gccgccgtct aaaaaggtga
     1261 tgtgtatttg agtaaaacag cttgcgtcat gcggtcgctg cgtatatgat gcgatgagta
     1321 aataaacaaa tacgcaaggg gaacgcatga aggttatcgc tgtacttaac cagaaaggcg
     1381 ggtcaggcaa gacgaccatc gcaacccatc tagcccgcgc cctgcaactc gccggggccg
     1441 atgttctgtt agtcgattcc gatccccagg gcagtgcccg cgattgggcg gccgtgcggg
     1501 aagatcaacc gctaaccgtt gtcggcatcg accgcccgac gattgaccgc gacgtgaagg
     1561 ccatcggccg gcgcgacttc gtagtgatcg acggagcgcc ccaggcggcg gacttggctg
     1621 tgtccgcgat caaggcagcc gacttcgtgc tgattccggt gcagccaagc ccttacgaca
     1681 tatgggccac cgccgacctg gtggagctgg ttaagcagcg cattgaggtc acggatggaa
     1741 ggctacaagc ggcctttgtc gtgtcgcggg cgatcaaagg cacgcgcatc ggcggtgagg
     1801 ttgccgaggc gctggccggg tacgagctgc ccattcttga gtcccgtatc acgcagcgcg
     1861 tgagctaccc aggcactgcc gccgccggca caaccgttct tgaatcagaa cccgagggcg
     1921 acgctgcccg cgaggtccag gcgctggccg ctgaaattaa atcaaaactc atttgagtta
     1981 atgaggtaaa gagaaaatga gcaaaagcac aaacacgcta agtgccggcc gtccgagcgc
     2041 acgcagcagc aaggctgcaa cgttggccag cctggcagac acgccagcca tgaagcgggt
     2101 caactttcag ttgccggcgg aggatcacac caagctgaag atgtacgcgg tacgccaagg
     2161 caagaccatt accgagctgc tatctgaata catcgcgcag ctaccagagt aaatgagcaa
     2221 atgaataaat gagtagatga attttagcgg ctaaaggagg cggcatggaa aatcaagaac
     2281 aaccaggcac cgacgccgtg gaatgcccca tgtgtggagg aacgggcggt tggccaggcg
     2341 taagcggctg ggttgcctgc cggccctgca atggcactgg aacccccaag cccgaggaat
     2401 cggcgtgagc ggtcgcaaac catccggccc ggtacaaatc ggcgcggcgc tgggtgatga
     2461 cctggtggag aagttgaagg ccgcgcaggc cgcccagcgg caacgcatcg aggcagaagc
     2521 acgccccggt gaatcgtggc aagcggccgc tgatcgaatc cgcaaagaat cccggcaacc
     2581 gccggcagcc ggtgcgccgt cgattaggaa gccgcccaag ggcgacgagc aaccagattt
     2641 tttcgttccg atgctctatg acgtgggcac ccgcgatagt cgcagcatca tggacgtggc
     2701 cgttttccgt ctgtcgaagc gtgaccgacg agctggcgag gtgatccgct acgagcttcc
     2761 agacgggcac gtagaggttt ccgcagggcc ggccggcatg gccagtgtgt gggattacga
     2821 cctggtactg atggcggttt cccatctaac cgaatccatg aaccgatacc gggaagggaa
     2881 gggagacaag cccggccgcg tgttccgtcc acacgttgcg gacgtactca agttctgccg
     2941 gcgagccgat ggcggaaagc agaaagacga cctggtagaa acctgcattc ggttaaacac
     3001 cacgcacgtt gccatgcagc gtacgaagaa ggccaagaac ggccgcctgg tgacggtatc
     3061 cgagggtgaa gccttgatta gccgctacaa gatcgtaaag agcgaaaccg ggcggccgga
     3121 gtacatcgag atcgagctag ctgattggat gtaccgcgag atcacagaag gcaagaaccc
     3181 ggacgtgctg acggttcacc ccgattactt tttgatcgat cccggcatcg gccgttttct
     3241 ctaccgcctg gcacgccgcg ccgcaggcaa ggcagaagcc agatggttgt tcaagacgat
     3301 ctacgaacgc agtggcagcg ccggagagtt caagaagttc tgtttcaccg tgcgcaagct
     3361 gatcgggtca aatgacctgc cggagtacga tttgaaggag gaggcggggc aggctggccc
     3421 gatcctagtc atgcgctacc gcaacctgat cgagggcgaa gcatccgccg gttcctaatg
     3481 tacggagcag atgctagggc aaattgccct agcaggggaa aaaggtcgaa aaggactctt
     3541 tcctgtggat agcacgtaca ttgggaaccc aaagccgtac attgggaacc ggaacccgta
     3601 cattgggaac ccaaagccgt acattgggaa ccggtcacac atgtaagtga ctgatataaa
     3661 agagaaaaaa ggcgattttt ccgcctaaaa ctctttaaaa cttattaaaa ctcttaaaac
     3721 ccgcctggcc tgtgcataac tgtctggcca gcgcacagcc gaagagctgc aaaaagcgcc
     3781 tacccttcgg tcgctgcgct ccctacgccc cgccgcttcg cgtcggccta tcgcggccgc
     3841 tggccgctca aaaatggctg gcctacggcc aggcaatcta ccagggcgcg gacaagccgc
     3901 gccgtcgcca ctcgaccgcc ggcgcccaca tcaaggcacc ctgcctcgcg cgtttcggtg
     3961 atgacggtga aaacctctga cacatgcagc tcccggtgac ggtcacagct tgtctgtaag
     4021 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     4081 gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc
     4141 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     4201 aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4261 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4321 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4381 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4441 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4501 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4561 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4621 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4681 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4741 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4801 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     4861 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4921 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4981 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5041 cgaaaactca cgttaaggga ttttggtcat gcattctagg tgattagaaa aactcatcga
     5101 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
     5161 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
     5221 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
     5281 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
     5341 gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
     5401 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagtcgaa
     5461 atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
     5521 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
     5581 atgctgtttt ccctgggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
     5641 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
     5701 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
     5761 gcttcccata caatcggtag attgtcgcac ctgattgccc gacattatcg cgagcccatt
     5821 tatacccata taaatcagca tccatgttgg aatttaatcg cggccttgag caagacgttt
     5881 cccgttgaat atggctcata acagaactta ttatttcctt cctcttttct acagtattta
     5941 aagatacccc aagaagctaa ttactgagag atcccctcat aatttcccca aagcgtaacc
     6001 atgtgtgaat aaattttgag ctagtagggt tgcagccacg agtaagtctt cccttgttat
     6061 tgtgtagcca gaatgccgca aaacttccat gcctaagcga actgttgaga gtacgtttcg
     6121 atttctgact gtgttagcct ggaagtgctt gtcccaacct tgtttctgag catgaacgcc
     6181 cgcaagccaa catgttagtt gaagcatcag ggcgattagc agcatgatat caaaacgctc
     6241 tgagctgctc gttcggctat ggcgtaggcc tagtccgtag gcaggacttt tcaagtctcg
     6301 gaaggtttct tcaatctgca ttcgcttcga atagatatta acaagttgtt tgggtgttcg
     6361 aatttcaaca ggtaagttag ttgctagaac ccatggctcc tttgccgacg ctgagtagat
     6421 tttaggtgac gggtggtgac aatgagtccg tgtcgagcgc tgattttttc ggcctttaga
     6481 gcgagattta tacaatagaa tttggcatga gattggattg cttttagtca gcctcttata
     6541 gcctaaagtc tttgagtgac tagatgacat atcatgtaag ttgctgatag gtttccagtt
     6601 ttccgctcct aggtctgcat attgtacttt tcctcttact cgacttaacc agtaccaacc
     6661 cagcttctca acggatttat accatggcac tttaaagcca gcatcactga caatgagcgg
     6721 tgtggtgtta ctcggtagaa tgctcgcaag gtcggctaga aattggtcat gagctttctt
     6781 tgaacattgc tctgaaagcg ggaacgcttt ctcataaaga gtaacagaac gaccgtgtag
     6841 tgcgactgaa gctcgcaata ccataagtcg tttttgctca cgaatatcag accagtcaac
     6901 aagtacaatg ggcatcgtat tgcccgaaca gataaagcta gcatgccaac ggtatacagc
     6961 gagtcgctct ttgtggaggt gacgattacc taacaatcgg tcgattcgtt tgatgttatg
     7021 ttttgttctc gctttggttg gcaggttacg gccaagttcg gtaagagtga gagttttaca
     7081 gtcaagtaat gcgtggcaag ccaacgttaa gctgttgagt cgttttaagt gtaattcggg
     7141 gcagaattgg taaagagagt cgtgtaaaat atcgagttcg cacatcttgt tgtctgatta
     7201 ttgatttttc gcgaaaccat ttgatcatat gacaagatgt gtatccacct taacttaatg
     7261 atttttacca aaatcattag gggattcatc agagctaatt ataacaagac gaactccaat
     7321 tcactgttcc ttgcattcta aaaccttaaa taccagaaaa cagctttttc aaagttgttt
     7381 tcaaagttgg cgtataacat agtatcgacg gagccgattt tgaaaccgcg gtgatcacag
     7441 gcagcaacgc tctgtcatcg ttacaatcaa catgctaccc tccgcgagat catccgtgtt
     7501 tcaaacccgg cagcttagtt gccgttcttc cgaatagcat cggtaacatg agcaaagtct
     7561 gccgccttac aacggctctc ccgctgacgc cgtcccggac tgatgggctg cctgtatcga
     7621 gtggtgattt tgtgccgagc tgccggtcgg ggagctgttg gctggctggt ggcaggatat
     7681 attgtggtgt aaacataacg aattcgtctc aggagagcga tgcgtctttt ccgatgaacc
     7741 gttccagcaa aaaagactaa gcgatgcgtc ttttccgctg aaccgttcca gcaaaaaaga
     7801 ctaagcgatg cgtcttttcc gctgaaccgt tccagcaaaa aagactagga gagcgatgcg
     7861 tcttttccgc tgaaccgttc cagcaaaaaa gactacttcg caagaccctt cctctatata
     7921 aggaagttca tttcatttgg agaggatact gtatttttac aacaattacc aacaacaaca
     7981 aacaacaaac aacattacaa ttactattta caattacaat ggtattcaca ctcgaagatt
     8041 tcgttgggga ctggcgacag acagccggct acaacctgga ccaagtcctt gaacagggag
     8101 gtgtgtccag tttgtttcag aatctcgggg tgtccgtaac tccgatccaa aggattgtcc
     8161 tgagcggtga aaatgggctg aagatcgaca tccatgtcat catcccgtat gaaggtctga
     8221 gcggcgacca aatgggccag atcgaaaaaa tttttaaggt ggtgtaccct gtggatgatc
     8281 atcactttaa ggtgatcctg cactatggca cactggtaat cgacggggtt acgccgaaca
     8341 tgatcgacta tttcggacgg ccgtatgaag gcatcgccgt gttcgacggc aaaaagatca
     8401 ctgtaacagg gaccctgtgg aacggcaaca aaattatcga cgagcgcctg atcaaccccg
     8461 acggctccct gctgttccga gtaaccatca acggagtgac cggctggcgg ctgtgcgaac
     8521 gcattctggc ttcggattat aaggaccatg acggagacta taaggaccat gacctcgacg
     8581 ctgcagcagc ggattataag gacgatgacg ataagtgagc ttctctagct agagtcgatc
     8641 gacaagctcg agtttctcca taataatgtg tgagtagttc ccagataagg gaattagggt
     8701 tcctataggg tttcgctcat gtgttgagca tataagaaac ccttagtatg tatttgtatt
     8761 tgtaaaatac ttctatcaat aaaatttcta attcctaaaa ccaaaatcca gtactaaaat
     8821 ccaga
//
