LOCUS       sequence-367784-        7865 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transcriptional unit for constitutive expression of CUP2:GAL4 driven
            by the nos promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7865)
  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 7865)
  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: pEPKK-alpha-1KN0253
FEATURES             Location/Qualifiers
     source          1..7865
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    365..389
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        487..666
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    702..757
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     polyA_signal    1726..1900
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    1920..1944
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3245..3874
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4303..5376
                     /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      5442..5636
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(5872..5894)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5980..6120
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6035..6054
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6135..6152)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6306..6894)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6386..6405)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6982..7797)
                     /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(6982..7797)
                     /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     7705..7724
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 aagacgaact ccaattcact gttccttgca ttctaaaacc ttaaatacca gaaaacagct
       61 ttttcaaagt tgttttcaaa gttggcgtat aacatagtat cgacggagcc gattttgaaa
      121 ccgcggtgat cacaggcagc aacgctctgt catcgttaca atcaacatgc taccctccgc
      181 gagatcatcc gtgtttcaaa cccggcagct tagttgccgt tcttccgaat agcatcggta
      241 acatgagcaa agtctgccgc cttacaacgg ctctcccgct gacgccgtcc cggactgatg
      301 ggctgcctgt atcgagtggt gattttgtgc cgagctgccg gtcggggagc tgttggctgg
      361 ctggtggcag gatatattgt ggtgtaaaca taacgaattc gtctcaggag agcggagaat
      421 taagggagtc acgttatgac ccccgccgat gacgcgggac aagccgtttt acgtttggaa
      481 ctgacagaac cgcaacgttg aaggagccac tcagccgcgg gtttctggag tttaatgagc
      541 taagcacata cgtcagaaac cattattgcg cgttcaaaag tcgcctaagg tcactatcag
      601 ctagcaaata tttcttgtca aaaatgctcc actgacgttc cataaattcc cctcggtatc
      661 caattagagt ctcatattca ctctcatact gtatttttac aacaattacc aacaacaaca
      721 aacaacaaac aacattacaa ttactattta caattacaat gatggtcgta attaacgggg
      781 tcaaatatgc ctgtgaaacg tgtatcaggg gtcacagggc ggcgcagtgt actcacactg
      841 atggtccgct acagatgatc agacgcaagg gaagaccatc gaccacatgt ggccattgta
      901 aagagctgag aagaaccaag aacttcaacc catccggtgg gtgcatgtgt gcctctgcac
      961 gacggccagc tgttggcagc aaggaagatg aaacacgatg tcgttgtgat gagggtgaac
     1021 cttgtaaatg tcataccaag aggaaaagca gccggaaatc aaagggaggg tcatgccaca
     1081 gaagggcaaa tgatgaagca gcgcatgtca atggtcttgg tattgcagat ctggacgttc
     1141 ttttgggcct aaatggtcgc tcgtcggatg tagacatgac aaccacattg ccgagtttga
     1201 agccacctct gcaaaacgga gaaattaagg ccgacagcat tgacaatctt gatttggctt
     1261 ccctcgatcc gcttgagcaa agccctagta tatctatgga acctgttagt atcaatgaaa
     1321 caggaagcgc atatacaact acgaacacag cactaaacga tattgacatt ccattctcca
     1381 tcaatgagtt gaacgagcta tacaaacaag tatcttcgca taactcacat tcacagccaa
     1441 tgacaaattc tcaagcgctt tcacaaccaa ttgcctcctc taacgttcat gataacttca
     1501 tgaataatga aatcacggct agtaaaattg atgatggtaa taattcaaaa ccactgtcac
     1561 ctggttggac ggaccaaact gcgtataacg cgtttggaat cactacaggg atgtttaata
     1621 ccactacaat ggatgatgta tataactatc tattcgatga tgaagatacc ccaccaaacc
     1681 caaaaaaaga ggcttctcta gctagagtcg atcgacaagc tcgagtttct ccataataat
     1741 gtgtgagtag ttcccagata agggaattag ggttcctata gggtttcgct catgtgttga
     1801 gcatataaga aacccttagt atgtatttgt atttgtaaaa tacttctatc aataaaattt
     1861 ctaattccta aaaccaaaat ccagtactaa aatccagatc gctgtcatga gacgaattct
     1921 gacaggatat attggcgggt aaacctaaga gaaaagagcg tttattagaa taatcggata
     1981 tttaaaaggg cgtgaaaagg tttatccgtt cgtccatttg tatgtgcatg ccaaccacag
     2041 ggttcccctc gggatcaaag tactttgatc caacccctcc gctgctatag tgcagtcggc
     2101 ttctgacgtt cagtgcagcc gtcatctgaa aacgacatgt cgcacaagtc ctaagttacg
     2161 cgacaggctg ccgccctgcc cttttcctgg cgttttcttg tcgcgtgttt tagtcgcata
     2221 aagtagaata cttgcgacta gaaccggaga cattacgcca tgaacaagag cgccgccgct
     2281 ggcctgctgg gctatgcccg cgtcagcacc gacgaccagg acttgaccaa ccaacgggcc
     2341 gaactgcacg cggccggctg caccaagctg ttttccgaga agatcaccgg caccaggcgc
     2401 gaccgcccgg agctggccag gatgcttgac cacctacgcc ctggcgacgt tgtgacagtg
     2461 accaggctag accgcctggc ccgcagcacc cgcgacctac tggacattgc cgagcgcatc
     2521 caggaggccg gcgcgggcct gcgtagcctg gcagagccgt gggccgacac caccacgccg
     2581 gccggccgca tggtgttgac cgtgttcgcc ggcattgccg agttcgagcg ttccctaatc
     2641 atcgaccgca cccggagcgg gcgcgaggcc gccaaggccc gaggcgtgaa gtttggcccc
     2701 cgccctaccc tcaccccggc acagatcgcg cacgcccgcg agctgatcga ccaggaaggc
     2761 cgcaccgtga aagaggcggc tgcactgctt ggcgtgcatc gctcgaccct gtaccgcgca
     2821 cttgagcgca gcgaggaagt gacgcccacc gaggccaggc ggcgcggtgc cttccgtgag
     2881 gacgcattga ccgaggccga cgccctggcg gccgccgaga atgaacgcca agaggaacaa
     2941 gcatgaaacc gcaccaggac ggccaggacg aaccgttttt cattaccgaa gagatcgagg
     3001 cggagatgat cgcggccggg tacgtgttcg agccgcccgc gcacctctca accgtgcggc
     3061 tgcatgaaat cctggccggt ttgtctgatg ccaagctggc ggcctggccg gccagcttgg
     3121 ccgctgaaga aaccgagcgc cgccgtctaa aaaggtgatg tgtatttgag taaaacagct
     3181 tgcgtcatgc ggtcgctgcg tatatgatgc gatgagtaaa taaacaaata cgcaagggga
     3241 acgcatgaag gttatcgctg tacttaacca gaaaggcggg tcaggcaaga cgaccatcgc
     3301 aacccatcta gcccgcgccc tgcaactcgc cggggccgat gttctgttag tcgattccga
     3361 tccccagggc agtgcccgcg attgggcggc cgtgcgggaa gatcaaccgc taaccgttgt
     3421 cggcatcgac cgcccgacga ttgaccgcga cgtgaaggcc atcggccggc gcgacttcgt
     3481 agtgatcgac ggagcgcccc aggcggcgga cttggctgtg tccgcgatca aggcagccga
     3541 cttcgtgctg attccggtgc agccaagccc ttacgacata tgggccaccg ccgacctggt
     3601 ggagctggtt aagcagcgca ttgaggtcac ggatggaagg ctacaagcgg cctttgtcgt
     3661 gtcgcgggcg atcaaaggca cgcgcatcgg cggtgaggtt gccgaggcgc tggccgggta
     3721 cgagctgccc attcttgagt cccgtatcac gcagcgcgtg agctacccag gcactgccgc
     3781 cgccggcaca accgttcttg aatcagaacc cgagggcgac gctgcccgcg aggtccaggc
     3841 gctggccgct gaaattaaat caaaactcat ttgagttaat gaggtaaaga gaaaatgagc
     3901 aaaagcacaa acacgctaag tgccggccgt ccgagcgcac gcagcagcaa ggctgcaacg
     3961 ttggccagcc tggcagacac gccagccatg aagcgggtca actttcagtt gccggcggag
     4021 gatcacacca agctgaagat gtacgcggta cgccaaggca agaccattac cgagctgcta
     4081 tctgaataca tcgcgcagct accagagtaa atgagcaaat gaataaatga gtagatgaat
     4141 tttagcggct aaaggaggcg gcatggaaaa tcaagaacaa ccaggcaccg acgccgtgga
     4201 atgccccatg tgtggaggaa cgggcggttg gccaggcgta agcggctggg ttgcctgccg
     4261 gccctgcaat ggcactggaa cccccaagcc cgaggaatcg gcgtgagcgg tcgcaaacca
     4321 tccggcccgg tacaaatcgg cgcggcgctg ggtgatgacc tggtggagaa gttgaaggcc
     4381 gcgcaggccg cccagcggca acgcatcgag gcagaagcac gccccggtga atcgtggcaa
     4441 gcggccgctg atcgaatccg caaagaatcc cggcaaccgc cggcagccgg tgcgccgtcg
     4501 attaggaagc cgcccaaggg cgacgagcaa ccagattttt tcgttccgat gctctatgac
     4561 gtgggcaccc gcgatagtcg cagcatcatg gacgtggccg ttttccgtct gtcgaagcgt
     4621 gaccgacgag ctggcgaggt gatccgctac gagcttccag acgggcacgt agaggtttcc
     4681 gcagggccgg ccggcatggc cagtgtgtgg gattacgacc tggtactgat ggcggtttcc
     4741 catctaaccg aatccatgaa ccgataccgg gaagggaagg gagacaagcc cggccgcgtg
     4801 ttccgtccac acgttgcgga cgtactcaag ttctgccggc gagccgatgg cggaaagcag
     4861 aaagacgacc tggtagaaac ctgcattcgg ttaaacacca cgcacgttgc catgcagcgt
     4921 acgaagaagg ccaagaacgg ccgcctggtg acggtatccg agggtgaagc cttgattagc
     4981 cgctacaaga tcgtaaagag cgaaaccggg cggccggagt acatcgagat cgagctagct
     5041 gattggatgt accgcgagat cacagaaggc aagaacccgg acgtgctgac ggttcacccc
     5101 gattactttt tgatcgatcc cggcatcggc cgttttctct accgcctggc acgccgcgcc
     5161 gcaggcaagg cagaagccag atggttgttc aagacgatct acgaacgcag tggcagcgcc
     5221 ggagagttca agaagttctg tttcaccgtg cgcaagctga tcgggtcaaa tgacctgccg
     5281 gagtacgatt tgaaggagga ggcggggcag gctggcccga tcctagtcat gcgctaccgc
     5341 aacctgatcg agggcgaagc atccgccggt tcctaatgta cggagcagat gctagggcaa
     5401 attgccctag caggggaaaa aggtcgaaaa ggactctttc ctgtggatag cacgtacatt
     5461 gggaacccaa agccgtacat tgggaaccgg aacccgtaca ttgggaaccc aaagccgtac
     5521 attgggaacc ggtcacacat gtaagtgact gatataaaag agaaaaaagg cgatttttcc
     5581 gcctaaaact ctttaaaact tattaaaact cttaaaaccc gcctggcctg tgcataactg
     5641 tctggccagc gcacagccga agagctgcaa aaagcgccta cccttcggtc gctgcgctcc
     5701 ctacgccccg ccgcttcgcg tcggcctatc gcggccgctg gccgctcaaa aatggctggc
     5761 ctacggccag gcaatctacc agggcgcgga caagccgcgc cgtcgccact cgaccgccgg
     5821 cgcccacatc aaggcaccct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     5881 catgcagctc ccggtgacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     5941 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg
     6001 tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat tgtactgaga
     6061 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     6121 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     6181 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     6241 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     6301 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     6361 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     6421 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     6481 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     6541 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     6601 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     6661 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     6721 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     6781 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     6841 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     6901 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     6961 ttggtcatgc attctaggtg attagaaaaa ctcatcgagc atcaaatgaa actgcaattt
     7021 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga
     7081 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac
     7141 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga
     7201 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt
     7261 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa
     7321 accgttattc attcgtgatt gcgcctgagc gagtcgaaat acgcgatcgc tgttaaaagg
     7381 acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat
     7441 attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc ctgggatcgc
     7501 agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg
     7561 cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct
     7621 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcggtagat
     7681 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc
     7741 catgttggaa tttaatcgcg gccttgagca agacgtttcc cgttgaatat ggctcataac
     7801 agaacttatt atttccttcc tcttttctac agtatttaaa gataccccaa gaagctaatt
     7861 ataac
//
