LOCUS       sequence-367809-        7697 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transcriptional unit for constitutive expression of nanoluc 
            luciferase driven by the 35s promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7697)
  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 7697)
  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-KN0005
FEATURES             Location/Qualifiers
     source          1..7697
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      396..590
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(826..848)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    934..1074
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     989..1008
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1089..1106)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1260..1848)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1340..1359)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1936..2751)
                     /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(1936..2751)
                     /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     2659..2678
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    3184..3208
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        3307..3652
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     3595..3615
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             3743..4255
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             4316..4339
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    4377..4551
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    4571..4595
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             5896..6525
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             join(6954..7697,1..330)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="VSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
ORIGIN
        1 atgtaccgcg agatcacaga aggcaagaac ccggacgtgc tgacggttca ccccgattac
       61 tttttgatcg atcccggcat cggccgtttt ctctaccgcc tggcacgccg cgccgcaggc
      121 aaggcagaag ccagatggtt gttcaagacg atctacgaac gcagtggcag cgccggagag
      181 ttcaagaagt tctgtttcac cgtgcgcaag ctgatcgggt caaatgacct gccggagtac
      241 gatttgaagg aggaggcggg gcaggctggc ccgatcctag tcatgcgcta ccgcaacctg
      301 atcgagggcg aagcatccgc cggttcctaa tgtacggagc agatgctagg gcaaattgcc
      361 ctagcagggg aaaaaggtcg aaaaggactc tttcctgtgg atagcacgta cattgggaac
      421 ccaaagccgt acattgggaa ccggaacccg tacattggga acccaaagcc gtacattggg
      481 aaccggtcac acatgtaagt gactgatata aaagagaaaa aaggcgattt ttccgcctaa
      541 aactctttaa aacttattaa aactcttaaa acccgcctgg cctgtgcata actgtctggc
      601 cagcgcacag ccgaagagct gcaaaaagcg cctacccttc ggtcgctgcg ctccctacgc
      661 cccgccgctt cgcgtcggcc tatcgcggcc gctggccgct caaaaatggc tggcctacgg
      721 ccaggcaatc taccagggcg cggacaagcc gcgccgtcgc cactcgaccg ccggcgccca
      781 catcaaggca ccctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
      841 gctcccggtg acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
      901 gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga
      961 tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac
     1021 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgctct
     1081 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     1141 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     1201 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     1261 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     1321 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     1381 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     1441 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     1501 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     1561 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     1621 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     1681 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     1741 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     1801 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     1861 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     1921 atgcattcta ggtgattaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
     1981 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
     2041 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
     2101 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
     2161 accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac
     2221 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
     2281 attcattcgt gattgcgcct gagcgagtcg aaatacgcga tcgctgttaa aaggacaatt
     2341 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
     2401 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttccctggga tcgcagtggt
     2461 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     2521 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
     2581 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcggt agattgtcgc
     2641 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     2701 ggaatttaat cgcggccttg agcaagacgt ttcccgttga atatggctca taacagaact
     2761 tattatttcc ttcctctttt ctacagtatt taaagatacc ccaagaagct aattataaca
     2821 agacgaactc caattcactg ttccttgcat tctaaaacct taaataccag aaaacagctt
     2881 tttcaaagtt gttttcaaag ttggcgtata acatagtatc gacggagccg attttgaaac
     2941 cgcggtgatc acaggcagca acgctctgtc atcgttacaa tcaacatgct accctccgcg
     3001 agatcatccg tgtttcaaac ccggcagctt agttgccgtt cttccgaata gcatcggtaa
     3061 catgagcaaa gtctgccgcc ttacaacggc tctcccgctg acgccgtccc ggactgatgg
     3121 gctgcctgta tcgagtggtg attttgtgcc gagctgccgg tcggggagct gttggctggc
     3181 tggtggcagg atatattgtg gtgtaaacat aacgaattcg tctcaggagg tcaacatggt
     3241 ggagcacgac actctggtct actccaaaaa tgtcaaagat acagtctcag aagatcaaag
     3301 ggctattgag acttttcaac aaaggataat ttcgggaaac ctcctcggat tccattgccc
     3361 agctatctgt cacttcatcg aaaggacagt agaaaaggaa ggtggctcct acaaatgcca
     3421 tcattgcgat aaaggaaagg ctatcattca agatctctct gccgacagtg gtcccaaaga
     3481 tggaccccca cccacgagga gcatcgtgga aaaagaagag gttccaacca cgtctacaaa
     3541 gcaagtggat tgatgtgaca tctccactga cgtaagggat gacgcacaat cccactatcc
     3601 ttcgcaagac ccttcctcta tataaggaag ttcatttcat ttggagagga cacgctcgag
     3661 tataagagct catttttaca acaattacca acaacaacaa acaacaaaca acattacaat
     3721 tacatttaca attatcgata caatggtatt cacactcgaa gatttcgttg gggactggcg
     3781 acagacagcc ggctacaacc tggaccaagt ccttgaacag ggaggtgtgt ccagtttgtt
     3841 tcagaatctc ggggtgtccg taactccgat ccaaaggatt gtcctgagcg gtgaaaatgg
     3901 gctgaagatc gacatccatg tcatcatccc gtatgaaggt ctgagcggcg accaaatggg
     3961 ccagatcgaa aaaattttta aggtggtgta ccctgtggat gatcatcact ttaaggtgat
     4021 cctgcactat ggcacactgg taatcgacgg ggttacgccg aacatgatcg actatttcgg
     4081 acggccgtat gaaggcatcg ccgtgttcga cggcaaaaag atcactgtaa cagggaccct
     4141 gtggaacggc aacaaaatta tcgacgagcg cctgatcaac cccgacggct ccctgctgtt
     4201 ccgagtaacc atcaacggag tgaccggctg gcggctgtgc gaacgcattc tggcttcgga
     4261 ttataaggac catgacggag actataagga ccatgacctc gacgctgcag cagcggatta
     4321 taaggacgat gacgataagt gagcttctct agctagagtc gatcgacaag ctcgagtttc
     4381 tccataataa tgtgtgagta gttcccagat aagggaatta gggttcctat agggtttcgc
     4441 tcatgtgttg agcatataag aaacccttag tatgtatttg tatttgtaaa atacttctat
     4501 caataaaatt tctaattcct aaaaccaaaa tccagtacta aaatccagat cgctgtcatg
     4561 agacgaattc tgacaggata tattggcggg taaacctaag agaaaagagc gtttattaga
     4621 ataatcggat atttaaaagg gcgtgaaaag gtttatccgt tcgtccattt gtatgtgcat
     4681 gccaaccaca gggttcccct cgggatcaaa gtactttgat ccaacccctc cgctgctata
     4741 gtgcagtcgg cttctgacgt tcagtgcagc cgtcatctga aaacgacatg tcgcacaagt
     4801 cctaagttac gcgacaggct gccgccctgc ccttttcctg gcgttttctt gtcgcgtgtt
     4861 ttagtcgcat aaagtagaat acttgcgact agaaccggag acattacgcc atgaacaaga
     4921 gcgccgccgc tggcctgctg ggctatgccc gcgtcagcac cgacgaccag gacttgacca
     4981 accaacgggc cgaactgcac gcggccggct gcaccaagct gttttccgag aagatcaccg
     5041 gcaccaggcg cgaccgcccg gagctggcca ggatgcttga ccacctacgc cctggcgacg
     5101 ttgtgacagt gaccaggcta gaccgcctgg cccgcagcac ccgcgaccta ctggacattg
     5161 ccgagcgcat ccaggaggcc ggcgcgggcc tgcgtagcct ggcagagccg tgggccgaca
     5221 ccaccacgcc ggccggccgc atggtgttga ccgtgttcgc cggcattgcc gagttcgagc
     5281 gttccctaat catcgaccgc acccggagcg ggcgcgaggc cgccaaggcc cgaggcgtga
     5341 agtttggccc ccgccctacc ctcaccccgg cacagatcgc gcacgcccgc gagctgatcg
     5401 accaggaagg ccgcaccgtg aaagaggcgg ctgcactgct tggcgtgcat cgctcgaccc
     5461 tgtaccgcgc acttgagcgc agcgaggaag tgacgcccac cgaggccagg cggcgcggtg
     5521 ccttccgtga ggacgcattg accgaggccg acgccctggc ggccgccgag aatgaacgcc
     5581 aagaggaaca agcatgaaac cgcaccagga cggccaggac gaaccgtttt tcattaccga
     5641 agagatcgag gcggagatga tcgcggccgg gtacgtgttc gagccgcccg cgcacctctc
     5701 aaccgtgcgg ctgcatgaaa tcctggccgg tttgtctgat gccaagctgg cggcctggcc
     5761 ggccagcttg gccgctgaag aaaccgagcg ccgccgtcta aaaaggtgat gtgtatttga
     5821 gtaaaacagc ttgcgtcatg cggtcgctgc gtatatgatg cgatgagtaa ataaacaaat
     5881 acgcaagggg aacgcatgaa ggttatcgct gtacttaacc agaaaggcgg gtcaggcaag
     5941 acgaccatcg caacccatct agcccgcgcc ctgcaactcg ccggggccga tgttctgtta
     6001 gtcgattccg atccccaggg cagtgcccgc gattgggcgg ccgtgcggga agatcaaccg
     6061 ctaaccgttg tcggcatcga ccgcccgacg attgaccgcg acgtgaaggc catcggccgg
     6121 cgcgacttcg tagtgatcga cggagcgccc caggcggcgg acttggctgt gtccgcgatc
     6181 aaggcagccg acttcgtgct gattccggtg cagccaagcc cttacgacat atgggccacc
     6241 gccgacctgg tggagctggt taagcagcgc attgaggtca cggatggaag gctacaagcg
     6301 gcctttgtcg tgtcgcgggc gatcaaaggc acgcgcatcg gcggtgaggt tgccgaggcg
     6361 ctggccgggt acgagctgcc cattcttgag tcccgtatca cgcagcgcgt gagctaccca
     6421 ggcactgccg ccgccggcac aaccgttctt gaatcagaac ccgagggcga cgctgcccgc
     6481 gaggtccagg cgctggccgc tgaaattaaa tcaaaactca tttgagttaa tgaggtaaag
     6541 agaaaatgag caaaagcaca aacacgctaa gtgccggccg tccgagcgca cgcagcagca
     6601 aggctgcaac gttggccagc ctggcagaca cgccagccat gaagcgggtc aactttcagt
     6661 tgccggcgga ggatcacacc aagctgaaga tgtacgcggt acgccaaggc aagaccatta
     6721 ccgagctgct atctgaatac atcgcgcagc taccagagta aatgagcaaa tgaataaatg
     6781 agtagatgaa ttttagcggc taaaggaggc ggcatggaaa atcaagaaca accaggcacc
     6841 gacgccgtgg aatgccccat gtgtggagga acgggcggtt ggccaggcgt aagcggctgg
     6901 gttgcctgcc ggccctgcaa tggcactgga acccccaagc ccgaggaatc ggcgtgagcg
     6961 gtcgcaaacc atccggcccg gtacaaatcg gcgcggcgct gggtgatgac ctggtggaga
     7021 agttgaaggc cgcgcaggcc gcccagcggc aacgcatcga ggcagaagca cgccccggtg
     7081 aatcgtggca agcggccgct gatcgaatcc gcaaagaatc ccggcaaccg ccggcagccg
     7141 gtgcgccgtc gattaggaag ccgcccaagg gcgacgagca accagatttt ttcgttccga
     7201 tgctctatga cgtgggcacc cgcgatagtc gcagcatcat ggacgtggcc gttttccgtc
     7261 tgtcgaagcg tgaccgacga gctggcgagg tgatccgcta cgagcttcca gacgggcacg
     7321 tagaggtttc cgcagggccg gccggcatgg ccagtgtgtg ggattacgac ctggtactga
     7381 tggcggtttc ccatctaacc gaatccatga accgataccg ggaagggaag ggagacaagc
     7441 ccggccgcgt gttccgtcca cacgttgcgg acgtactcaa gttctgccgg cgagccgatg
     7501 gcggaaagca gaaagacgac ctggtagaaa cctgcattcg gttaaacacc acgcacgttg
     7561 ccatgcagcg tacgaagaag gccaagaacg gccgcctggt gacggtatcc gagggtgaag
     7621 ccttgattag ccgctacaag atcgtaaaga gcgaaaccgg gcggccggag tacatcgaga
     7681 tcgagctagc tgattgg
//
