LOCUS       sequence-447734-        9121 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transcriptional unit for the copper and PhiC31 regulated expression 
            of BeYDV Rep/RepA..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9121)
  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 9121)
  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_alpha1_CBS-att-OCS-BeYDVRep-RepA-tNos (G...
FEATURES             Location/Qualifiers
     source          1..9121
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      362..1068
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     protein_bind    1077..1110
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     terminator      2277..2529
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(2564..2582)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    2770..2794
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             4095..4724
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             5153..6226
                     /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      6292..6486
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(6722..6744)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6830..6970
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6885..6904
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6985..7002)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7156..7744)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7236..7255)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7832..8647)
                     /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(7832..8647)
                     /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     8555..8574
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    9080..9104
                     /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 acttgtgccc caactggggt aacctttgag ttctctcagt tgggggcgta
      361 gctgctttaa tgagatatgc gagaagccta tgatcgcatg atatttgctt tcaattctgt
      421 tgtgcacgtt gtaaaaaacc tgagcatgtg tagctcagat ccttaccgcc ggtttcggtt
      481 cattctaatg aatatatcac cgttactatc gtatttttat gaataatatt ctccgttcaa
      541 tttactgatt gtaccctact acttatatgt acaatattaa aatgaaaaca atatattgtg
      601 ctgaataggt ttatagcgac atctatgata gagcgccaca ataacaaaca attgcgtttt
      661 attattacaa atccaatttt aaaaaaagcg gcagaaccgg tcaaacctaa aagactgatt
      721 acataaatct tattcaaatt tcaaaagtgc cccaggggct agtatctacg acacaccgag
      781 cggcgaacta ataacgctca ctgaagggaa ctccggttcc ccgccggcgc gcatgggtga
      841 gattccttga agttgagtat tggccgtccg ctctaccgaa agttacgggc accattcaac
      901 ccggtccagc acggcggccg ggtaaccgac ttgctgcccc gagaattatg cagcattttt
      961 ttggtgtatg tgggccccaa atgaagtgca ggtcaaacct tgacagtgac gacaaatcgt
     1021 tgggcgggtc cagggcgaat tttgcgacaa catgtcgagg ctcagcagcg gtgcgggtgc
     1081 cagggcgtgc ccttgggctc cccgggcgcg tactccacaa ggttaaattt gctgactctt
     1141 ttttaccttt catttctgaa aaatgccttc tgctagtaag aacttcagac tccaatctaa
     1201 atatgttttc cttacctatc ccaagtgctc atctcaaaga gatgatttat tccagtttct
     1261 ctgggagaaa ctcacacctt ttcttatttt cttccttggt gttgcttctg agcttcatca
     1321 agatggcact acccactatc atgctcttct ccagcttgat aaaaaacctt gtattaggga
     1381 tccttctttt ttcgattttg aaggaaatca ccctaatatc cagccagcta gaaactctaa
     1441 acaagtcctt gattacatat caaaggacgg agatattaaa accagaggag atttccgaga
     1501 tcataaggtt tctcctcgca aatctgacgc acgatggaga actattatcc agactgcaac
     1561 gtctaaggag gaatatcttg acatgatcaa ggaagaattc cctcatgaat gggcaacaaa
     1621 gcttcaatgg ctggaatatt cagccaacaa attattccct ccacaacctg aaccgtatgt
     1681 gtcgcccttc acagaatcag atcttcgctg ccacgaagat ctacactcct ggagggaaac
     1741 ccatctatac catgtaagca tagacgctta tacttacata catcctgtct cataccaaca
     1801 agctcaatct gaccttgaat ggatggccga tttaaccagg acaatggaag gaatggaatc
     1861 cgacacccca gcctctacat ctgcggacca actcgtaccg gaaagaccac ctgggctaga
     1921 agtctcggac gacacaacta ttggaacggt accatcgatt tcaccaacta cgatgaacac
     1981 gccacctata atatcatcga cgacatcccc ttcaagttcg tcccattgtg gaagcaatta
     2041 ataggttgcc agtctgattt cactgtcaac cctaaatatg gaaaaaagaa gaaaataaaa
     2101 ggtgggatcc cttctataat tctttgcaat cctgacgaag actggatgtt atcaatgaca
     2161 agtcaacaga aggattactt taaagataat tgcgtcaccc actacatgtg tgacggggag
     2221 actttttttg ctcgggaatc gtcgagtcac tgagcttgga atggatcttc gatcccgatc
     2281 gttcaaacat ttggcaataa agtttcttaa gattgaatcc tgttgccggt cttgcgacga
     2341 ttatcatata atttctgttg aattacgtta agcatgtaat aattaacatg taatgcatga
     2401 cgttatttat gagatgggtt tttatgatta gagtcccgca attatacatt taatacgcga
     2461 tagaaaacaa aatatagcgc gcaaactagg ataaattatc gcgcgcggtg tcatctatgt
     2521 tactagatcg ggaattgcca agctaattct tgaagacgaa agggcctcgt gatacgccta
     2581 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     2641 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     2701 ctcatgagac aataaccctg ataaatgctt caataatggg accgactcgc gctgtcatga
     2761 gacgaattct gacaggatat attggcgggt aaacctaaga gaaaagagcg tttattagaa
     2821 taatcggata tttaaaaggg cgtgaaaagg tttatccgtt cgtccatttg tatgtgcatg
     2881 ccaaccacag ggttcccctc gggatcaaag tactttgatc caacccctcc gctgctatag
     2941 tgcagtcggc ttctgacgtt cagtgcagcc gtcatctgaa aacgacatgt cgcacaagtc
     3001 ctaagttacg cgacaggctg ccgccctgcc cttttcctgg cgttttcttg tcgcgtgttt
     3061 tagtcgcata aagtagaata cttgcgacta gaaccggaga cattacgcca tgaacaagag
     3121 cgccgccgct ggcctgctgg gctatgcccg cgtcagcacc gacgaccagg acttgaccaa
     3181 ccaacgggcc gaactgcacg cggccggctg caccaagctg ttttccgaga agatcaccgg
     3241 caccaggcgc gaccgcccgg agctggccag gatgcttgac cacctacgcc ctggcgacgt
     3301 tgtgacagtg accaggctag accgcctggc ccgcagcacc cgcgacctac tggacattgc
     3361 cgagcgcatc caggaggccg gcgcgggcct gcgtagcctg gcagagccgt gggccgacac
     3421 caccacgccg gccggccgca tggtgttgac cgtgttcgcc ggcattgccg agttcgagcg
     3481 ttccctaatc atcgaccgca cccggagcgg gcgcgaggcc gccaaggccc gaggcgtgaa
     3541 gtttggcccc cgccctaccc tcaccccggc acagatcgcg cacgcccgcg agctgatcga
     3601 ccaggaaggc cgcaccgtga aagaggcggc tgcactgctt ggcgtgcatc gctcgaccct
     3661 gtaccgcgca cttgagcgca gcgaggaagt gacgcccacc gaggccaggc ggcgcggtgc
     3721 cttccgtgag gacgcattga ccgaggccga cgccctggcg gccgccgaga atgaacgcca
     3781 agaggaacaa gcatgaaacc gcaccaggac ggccaggacg aaccgttttt cattaccgaa
     3841 gagatcgagg cggagatgat cgcggccggg tacgtgttcg agccgcccgc gcacctctca
     3901 accgtgcggc tgcatgaaat cctggccggt ttgtctgatg ccaagctggc ggcctggccg
     3961 gccagcttgg ccgctgaaga aaccgagcgc cgccgtctaa aaaggtgatg tgtatttgag
     4021 taaaacagct tgcgtcatgc ggtcgctgcg tatatgatgc gatgagtaaa taaacaaata
     4081 cgcaagggga acgcatgaag gttatcgctg tacttaacca gaaaggcggg tcaggcaaga
     4141 cgaccatcgc aacccatcta gcccgcgccc tgcaactcgc cggggccgat gttctgttag
     4201 tcgattccga tccccagggc agtgcccgcg attgggcggc cgtgcgggaa gatcaaccgc
     4261 taaccgttgt cggcatcgac cgcccgacga ttgaccgcga cgtgaaggcc atcggccggc
     4321 gcgacttcgt agtgatcgac ggagcgcccc aggcggcgga cttggctgtg tccgcgatca
     4381 aggcagccga cttcgtgctg attccggtgc agccaagccc ttacgacata tgggccaccg
     4441 ccgacctggt ggagctggtt aagcagcgca ttgaggtcac ggatggaagg ctacaagcgg
     4501 cctttgtcgt gtcgcgggcg atcaaaggca cgcgcatcgg cggtgaggtt gccgaggcgc
     4561 tggccgggta cgagctgccc attcttgagt cccgtatcac gcagcgcgtg agctacccag
     4621 gcactgccgc cgccggcaca accgttcttg aatcagaacc cgagggcgac gctgcccgcg
     4681 aggtccaggc gctggccgct gaaattaaat caaaactcat ttgagttaat gaggtaaaga
     4741 gaaaatgagc aaaagcacaa acacgctaag tgccggccgt ccgagcgcac gcagcagcaa
     4801 ggctgcaacg ttggccagcc tggcagacac gccagccatg aagcgggtca actttcagtt
     4861 gccggcggag gatcacacca agctgaagat gtacgcggta cgccaaggca agaccattac
     4921 cgagctgcta tctgaataca tcgcgcagct accagagtaa atgagcaaat gaataaatga
     4981 gtagatgaat tttagcggct aaaggaggcg gcatggaaaa tcaagaacaa ccaggcaccg
     5041 acgccgtgga atgccccatg tgtggaggaa cgggcggttg gccaggcgta agcggctggg
     5101 ttgcctgccg gccctgcaat ggcactggaa cccccaagcc cgaggaatcg gcgtgagcgg
     5161 tcgcaaacca tccggcccgg tacaaatcgg cgcggcgctg ggtgatgacc tggtggagaa
     5221 gttgaaggcc gcgcaggccg cccagcggca acgcatcgag gcagaagcac gccccggtga
     5281 atcgtggcaa gcggccgctg atcgaatccg caaagaatcc cggcaaccgc cggcagccgg
     5341 tgcgccgtcg attaggaagc cgcccaaggg cgacgagcaa ccagattttt tcgttccgat
     5401 gctctatgac gtgggcaccc gcgatagtcg cagcatcatg gacgtggccg ttttccgtct
     5461 gtcgaagcgt gaccgacgag ctggcgaggt gatccgctac gagcttccag acgggcacgt
     5521 agaggtttcc gcagggccgg ccggcatggc cagtgtgtgg gattacgacc tggtactgat
     5581 ggcggtttcc catctaaccg aatccatgaa ccgataccgg gaagggaagg gagacaagcc
     5641 cggccgcgtg ttccgtccac acgttgcgga cgtactcaag ttctgccggc gagccgatgg
     5701 cggaaagcag aaagacgacc tggtagaaac ctgcattcgg ttaaacacca cgcacgttgc
     5761 catgcagcgt acgaagaagg ccaagaacgg ccgcctggtg acggtatccg agggtgaagc
     5821 cttgattagc cgctacaaga tcgtaaagag cgaaaccggg cggccggagt acatcgagat
     5881 cgagctagct gattggatgt accgcgagat cacagaaggc aagaacccgg acgtgctgac
     5941 ggttcacccc gattactttt tgatcgatcc cggcatcggc cgttttctct accgcctggc
     6001 acgccgcgcc gcaggcaagg cagaagccag atggttgttc aagacgatct acgaacgcag
     6061 tggcagcgcc ggagagttca agaagttctg tttcaccgtg cgcaagctga tcgggtcaaa
     6121 tgacctgccg gagtacgatt tgaaggagga ggcggggcag gctggcccga tcctagtcat
     6181 gcgctaccgc aacctgatcg agggcgaagc atccgccggt tcctaatgta cggagcagat
     6241 gctagggcaa attgccctag caggggaaaa aggtcgaaaa ggactctttc ctgtggatag
     6301 cacgtacatt gggaacccaa agccgtacat tgggaaccgg aacccgtaca ttgggaaccc
     6361 aaagccgtac attgggaacc ggtcacacat gtaagtgact gatataaaag agaaaaaagg
     6421 cgatttttcc gcctaaaact ctttaaaact tattaaaact cttaaaaccc gcctggcctg
     6481 tgcataactg tctggccagc gcacagccga agagctgcaa aaagcgccta cccttcggtc
     6541 gctgcgctcc ctacgccccg ccgcttcgcg tcggcctatc gcggccgctg gccgctcaaa
     6601 aatggctggc ctacggccag gcaatctacc agggcgcgga caagccgcgc cgtcgccact
     6661 cgaccgccgg cgcccacatc aaggcaccct gcctcgcgcg tttcggtgat gacggtgaaa
     6721 acctctgaca catgcagctc ccggtgacgg tcacagcttg tctgtaagcg gatgccggga
     6781 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga
     6841 cccagtcacg tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat
     6901 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     6961 ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     7021 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     7081 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     7141 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     7201 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     7261 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     7321 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     7381 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     7441 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     7501 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     7561 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     7621 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     7681 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     7741 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     7801 ttaagggatt ttggtcatgc attctaggtg attagaaaaa ctcatcgagc atcaaatgaa
     7861 actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta
     7921 atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg
     7981 cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt
     8041 tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat
     8101 gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg
     8161 catcaaccaa accgttattc attcgtgatt gcgcctgagc gagtcgaaat acgcgatcgc
     8221 tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg
     8281 catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc
     8341 ctgggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg
     8401 tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat
     8461 tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca
     8521 atcggtagat tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata
     8581 aatcagcatc catgttggaa tttaatcgcg gccttgagca agacgtttcc cgttgaatat
     8641 ggctcataac agaacttatt atttccttcc tcttttctac agtatttaaa gataccccaa
     8701 gaagctaatt ataacaagac gaactccaat tcactgttcc ttgcattcta aaaccttaaa
     8761 taccagaaaa cagctttttc aaagttgttt tcaaagttgg cgtataacat agtatcgacg
     8821 gagccgattt tgaaaccgcg gtgatcacag gcagcaacgc tctgtcatcg ttacaatcaa
     8881 catgctaccc tccgcgagat catccgtgtt tcaaacccgg cagcttagtt gccgttcttc
     8941 cgaatagcat cggtaacatg agcaaagtct gccgccttac aacggctctc ccgctgacgc
     9001 cgtcccggac tgatgggctg cctgtatcga gtggtgattt tgtgccgagc tgccggtcgg
     9061 ggagctgttg gctggctggt ggcaggatat attgtggtgt aaacataacg aattcgtctc
     9121 a
//
