LOCUS       sequence-447737-        8317 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transcriptional unit for copper-regulation of BeYDV Rep/RepA with 
            4xCBS domain+minimal promoter DFR for induction by copper..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8317)
  AUTHORS   Garcia-Perez E, Vazquez-Vilar M, Lozano-Duran R, Orzaez D
  TITLE     CuBe: a geminivirus-based copper-regulated expression system 
            suitable for post-harvest activation.
  JOURNAL   Plant Biotechnol J. 2025 Jan;23(1):141-155. doi: 10.1111/pbi.14485. 
            Epub 2024 Oct 22.
  PUBMED    39435699
REFERENCE   2  (bases 1 to 8317)
  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"; doi: "10.1111/pbi.14485";
            journalName: "Plant Biotechnol J"; date: "2025-01"; volume: "23"; 
            issue: "1"; pages: "141-155"
COMMENT     Sequence Label: pDGB3_alpha2_4xCBS-mDFR-Rep-RepA-tNOS (GB3625)
FEATURES             Location/Qualifiers
     source          1..8317
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1472..1724
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(1759..1777)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    1961..1985
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3286..3915
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4344..5417
                     /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      5483..5677
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(5913..5935)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6021..6161
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6076..6095
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6176..6193)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6347..6935)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6427..6446)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7023..7838)
                     /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(7023..7838)
                     /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     7746..7765
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    8271..8295
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 ggagagtggt cttgggagag cgatgcgtct tttccgatga accgttccag caaaaaagac
       61 taagcgatgc gtcttttccg ctgaaccgtt ccagcaaaaa agactaagcg atgcgtcttt
      121 tccgctgaac cgttccagca aaaaagacta ggagagcgat gcgtcttttc cgctgaaccg
      181 ttccagcaaa aaagactatc ccgtcctaat acctctacgt gcaagcagta taaaaacaaa
      241 atacataaac ataactaaaa tctcatatcg ttgtaattgt catatactag tggtaattaa
      301 gtccagttac acttaaggtt aaatttgctg actctttttt acctttcatt tctgaaaatg
      361 ccttctgcta gtaagaactt cagactccaa tctaaatatg ttttccttac ctatcccaag
      421 tgctcatctc aaagagatga tttattccag tttctctggg agaaactcac accttttctt
      481 attttcttcc ttggtgttgc ttctgagctt catcaagatg gcactaccca ctatcatgct
      541 cttctccagc ttgataaaaa accttgtatt agggatcctt cttttttcga ttttgaagga
      601 aatcacccta atatccagcc agctagaaac tctaaacaag tccttgatta catatcaaag
      661 gacggagata ttaaaaccag aggagatttc cgagatcata aggtttctcc tcgcaaatct
      721 gacgcacgat ggagaactat tatccagact gcaacgtcta aggaggaata tcttgacatg
      781 atcaaggaag aattccctca tgaatgggca acaaagcttc aatggctgga atattcagcc
      841 aacaaattat tccctccaca acctgaaccg tatgtgtcgc ccttcacaga atcagatctt
      901 cgctgccacg aagatctaca ctcctggagg gaaacccatc tataccatgt aagcatagac
      961 gcttatactt acatacatcc tgtctcatac caacaagctc aatctgacct tgaatggatg
     1021 gccgatttaa ccaggacaat ggaaggaatg gaatccgaca ccccagcctc tacatctgcg
     1081 gaccaactcg taccggaaag accacctggg ctagaagtct cggacgacac aactattgga
     1141 acggtaccat cgatttcacc aactacgatg aacacgccac ctataatatc atcgacgaca
     1201 tccccttcaa gttcgtccca ttgtggaagc aattaatagg ttgccagtct gatttcactg
     1261 tcaaccctaa atatggaaaa aagaagaaaa taaaaggtgg gatcccttct ataattcttt
     1321 gcaatcctga cgaagactgg atgttatcaa tgacaagtca acagaaggat tactttaaag
     1381 ataattgcgt cacccactac atgtgtgacg gggagacttt ttttgctcgg gaatcgtcga
     1441 gtcactgagc ttggaatgga tcttcgatcc cgatcgttca aacatttggc aataaagttt
     1501 cttaagattg aatcctgttg ccggtcttgc gacgattatc atataatttc tgttgaatta
     1561 cgttaagcat gtaataatta acatgtaatg catgacgtta tttatgagat gggtttttat
     1621 gattagagtc ccgcaattat acatttaata cgcgatagaa aacaaaatat agcgcgcaaa
     1681 ctaggataaa ttatcgcgcg cggtgtcatc tatgttacta gatcgggaat tgccaagcta
     1741 attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     1801 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     1861 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     1921 tgcttcaata atgggaccga ctcgcgcttg agacgaagct tgacaggata tattggcggg
     1981 taaacctaag agaaaagagc gtttattaga ataatcggat atttaaaagg gcgtgaaaag
     2041 gtttatccgt tcgtccattt gtatgtgcat gccaaccaca gggttcccct cgggatcaaa
     2101 gtactttgat ccaacccctc cgctgctata gtgcagtcgg cttctgacgt tcagtgcagc
     2161 cgtcatctga aaacgacatg tcgcacaagt cctaagttac gcgacaggct gccgccctgc
     2221 ccttttcctg gcgttttctt gtcgcgtgtt ttagtcgcat aaagtagaat acttgcgact
     2281 agaaccggag acattacgcc atgaacaaga gcgccgccgc tggcctgctg ggctatgccc
     2341 gcgtcagcac cgacgaccag gacttgacca accaacgggc cgaactgcac gcggccggct
     2401 gcaccaagct gttttccgag aagatcaccg gcaccaggcg cgaccgcccg gagctggcca
     2461 ggatgcttga ccacctacgc cctggcgacg ttgtgacagt gaccaggcta gaccgcctgg
     2521 cccgcagcac ccgcgaccta ctggacattg ccgagcgcat ccaggaggcc ggcgcgggcc
     2581 tgcgtagcct ggcagagccg tgggccgaca ccaccacgcc ggccggccgc atggtgttga
     2641 ccgtgttcgc cggcattgcc gagttcgagc gttccctaat catcgaccgc acccggagcg
     2701 ggcgcgaggc cgccaaggcc cgaggcgtga agtttggccc ccgccctacc ctcaccccgg
     2761 cacagatcgc gcacgcccgc gagctgatcg accaggaagg ccgcaccgtg aaagaggcgg
     2821 ctgcactgct tggcgtgcat cgctcgaccc tgtaccgcgc acttgagcgc agcgaggaag
     2881 tgacgcccac cgaggccagg cggcgcggtg ccttccgtga ggacgcattg accgaggccg
     2941 acgccctggc ggccgccgag aatgaacgcc aagaggaaca agcatgaaac cgcaccagga
     3001 cggccaggac gaaccgtttt tcattaccga agagatcgag gcggagatga tcgcggccgg
     3061 gtacgtgttc gagccgcccg cgcacctctc aaccgtgcgg ctgcatgaaa tcctggccgg
     3121 tttgtctgat gccaagctgg cggcctggcc ggccagcttg gccgctgaag aaaccgagcg
     3181 ccgccgtcta aaaaggtgat gtgtatttga gtaaaacagc ttgcgtcatg cggtcgctgc
     3241 gtatatgatg cgatgagtaa ataaacaaat acgcaagggg aacgcatgaa ggttatcgct
     3301 gtacttaacc agaaaggcgg gtcaggcaag acgaccatcg caacccatct agcccgcgcc
     3361 ctgcaactcg ccggggccga tgttctgtta gtcgattccg atccccaggg cagtgcccgc
     3421 gattgggcgg ccgtgcggga agatcaaccg ctaaccgttg tcggcatcga ccgcccgacg
     3481 attgaccgcg acgtgaaggc catcggccgg cgcgacttcg tagtgatcga cggagcgccc
     3541 caggcggcgg acttggctgt gtccgcgatc aaggcagccg acttcgtgct gattccggtg
     3601 cagccaagcc cttacgacat atgggccacc gccgacctgg tggagctggt taagcagcgc
     3661 attgaggtca cggatggaag gctacaagcg gcctttgtcg tgtcgcgggc gatcaaaggc
     3721 acgcgcatcg gcggtgaggt tgccgaggcg ctggccgggt acgagctgcc cattcttgag
     3781 tcccgtatca cgcagcgcgt gagctaccca ggcactgccg ccgccggcac aaccgttctt
     3841 gaatcagaac ccgagggcga cgctgcccgc gaggtccagg cgctggccgc tgaaattaaa
     3901 tcaaaactca tttgagttaa tgaggtaaag agaaaatgag caaaagcaca aacacgctaa
     3961 gtgccggccg tccgagcgca cgcagcagca aggctgcaac gttggccagc ctggcagaca
     4021 cgccagccat gaagcgggtc aactttcagt tgccggcgga ggatcacacc aagctgaaga
     4081 tgtacgcggt acgccaaggc aagaccatta ccgagctgct atctgaatac atcgcgcagc
     4141 taccagagta aatgagcaaa tgaataaatg agtagatgaa ttttagcggc taaaggaggc
     4201 ggcatggaaa atcaagaaca accaggcacc gacgccgtgg aatgccccat gtgtggagga
     4261 acgggcggtt ggccaggcgt aagcggctgg gttgcctgcc ggccctgcaa tggcactgga
     4321 acccccaagc ccgaggaatc ggcgtgagcg gtcgcaaacc atccggcccg gtacaaatcg
     4381 gcgcggcgct gggtgatgac ctggtggaga agttgaaggc cgcgcaggcc gcccagcggc
     4441 aacgcatcga ggcagaagca cgccccggtg aatcgtggca agcggccgct gatcgaatcc
     4501 gcaaagaatc ccggcaaccg ccggcagccg gtgcgccgtc gattaggaag ccgcccaagg
     4561 gcgacgagca accagatttt ttcgttccga tgctctatga cgtgggcacc cgcgatagtc
     4621 gcagcatcat ggacgtggcc gttttccgtc tgtcgaagcg tgaccgacga gctggcgagg
     4681 tgatccgcta cgagcttcca gacgggcacg tagaggtttc cgcagggccg gccggcatgg
     4741 ccagtgtgtg ggattacgac ctggtactga tggcggtttc ccatctaacc gaatccatga
     4801 accgataccg ggaagggaag ggagacaagc ccggccgcgt gttccgtcca cacgttgcgg
     4861 acgtactcaa gttctgccgg cgagccgatg gcggaaagca gaaagacgac ctggtagaaa
     4921 cctgcattcg gttaaacacc acgcacgttg ccatgcagcg tacgaagaag gccaagaacg
     4981 gccgcctggt gacggtatcc gagggtgaag ccttgattag ccgctacaag atcgtaaaga
     5041 gcgaaaccgg gcggccggag tacatcgaga tcgagctagc tgattggatg taccgcgaga
     5101 tcacagaagg caagaacccg gacgtgctga cggttcaccc cgattacttt ttgatcgatc
     5161 ccggcatcgg ccgttttctc taccgcctgg cacgccgcgc cgcaggcaag gcagaagcca
     5221 gatggttgtt caagacgatc tacgaacgca gtggcagcgc cggagagttc aagaagttct
     5281 gtttcaccgt gcgcaagctg atcgggtcaa atgacctgcc ggagtacgat ttgaaggagg
     5341 aggcggggca ggctggcccg atcctagtca tgcgctaccg caacctgatc gagggcgaag
     5401 catccgccgg ttcctaatgt acggagcaga tgctagggca aattgcccta gcaggggaaa
     5461 aaggtcgaaa aggactcttt cctgtggata gcacgtacat tgggaaccca aagccgtaca
     5521 ttgggaaccg gaacccgtac attgggaacc caaagccgta cattgggaac cggtcacaca
     5581 tgtaagtgac tgatataaaa gagaaaaaag gcgatttttc cgcctaaaac tctttaaaac
     5641 ttattaaaac tcttaaaacc cgcctggcct gtgcataact gtctggccag cgcacagccg
     5701 aagagctgca aaaagcgcct acccttcggt cgctgcgctc cctacgcccc gccgcttcgc
     5761 gtcggcctat cgcggccgct ggccgctcaa aaatggctgg cctacggcca ggcaatctac
     5821 cagggcgcgg acaagccgcg ccgtcgccac tcgaccgccg gcgcccacat caaggcaccc
     5881 tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggtgacg
     5941 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     6001 ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat
     6061 actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     6121 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc
     6181 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     6241 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     6301 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     6361 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     6421 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6481 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6541 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6601 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6661 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6721 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6781 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6841 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     6901 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6961 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg cattctaggt
     7021 gattagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
     7081 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     7141 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     7201 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     7261 actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc
     7321 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     7381 tgcgcctgag cgagtcgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
     7441 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     7501 tattcttcta atacctggaa tgctgttttc cctgggatcg cagtggtgag taaccatgca
     7561 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     7621 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     7681 aacaactctg gcgcatcggg cttcccatac aatcggtaga ttgtcgcacc tgattgcccg
     7741 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     7801 ggccttgagc aagacgtttc ccgttgaata tggctcataa cagaacttat tatttccttc
     7861 ctcttttcta cagtatttaa agatacccca agaagctaat tataacaaga cgaactccaa
     7921 ttcactgttc cttgcattct aaaaccttaa ataccagaaa acagcttttt caaagttgtt
     7981 ttcaaagttg gcgtataaca tagtatcgac ggagccgatt ttgaaaccgc ggtgatcaca
     8041 ggcagcaacg ctctgtcatc gttacaatca acatgctacc ctccgcgaga tcatccgtgt
     8101 ttcaaacccg gcagcttagt tgccgttctt ccgaatagca tcggtaacat gagcaaagtc
     8161 tgccgcctta caacggctct cccgctgacg ccgtcccgga ctgatgggct gcctgtatcg
     8221 agtggtgatt ttgtgccgag ctgccggtcg gggagctgtt ggctggctgg tggcaggata
     8281 tattgtggtg taaacataac aagcttcgtc tcagtca
//
