LOCUS       sequence-119066-        8421 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transcriptional unit for Tomato bushy stunt virus (TBSV) silencing 
            suppressor P19 plant expression driven by the 35S promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8421)
  AUTHORS   Sarrion-Perdigones A, Vazquez-Vilar M, Palaci J, Castelijns B, 
            Forment J, Ziarsolo P, Blanca J, Granell A, Orzaez D
  TITLE     GoldenBraid 2.0: a comprehensive DNA assembly framework for plant 
            synthetic biology.
  JOURNAL   Plant Physiol. 2013 Jul;162(3):1618-31.
  PUBMED    23669743
REFERENCE   2  (bases 1 to 8421)
  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 
            Physiol."; date: "2013-07"; volume: "162(3)"; pages: "1618-31"
COMMENT     Sequence Label: pDGB3alpha2_35S:P19:Tnos  (GB1203)
FEATURES             Location/Qualifiers
     source          1..8421
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        616..960
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     904..924
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      1576..1828
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(1863..1881)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    2066..2090
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3391..4020
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIGTHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4454..5521
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     rep_origin      5587..5781
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(6017..6039)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6125..6265
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6180..6199
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6280..6297)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6451..7039)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6531..6550)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7127..7942)
                     /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(7127..7942)
                     /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     7850..7869
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    8375..8399
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 ggagactaga gccaagctga tctcctttgc cccggagatc accatggacg actttctcta
       61 tctctacgat ctaggaagaa agttcgacgg agaaggtgac gataccatgt tcaccaccga
      121 taatgagaag attagcctct tcaatttcag aaagaatgct gacccacaga tggttagaga
      181 ggcctacgcg gcaggtctga tcaagacgat ctacccgagt aataatctcc aggagatcaa
      241 ataccttccc aagaaggtta aagatgcagt caaaagattc aggactaact gcatcaagaa
      301 cacagagaaa gatatatttc tcaagatcag aagtactatt ccagtatgga cgattcaagg
      361 cttgcttcat aaaccaaggc aagtaataga gattggagtc tctaagaaag tagttcctac
      421 tgaatcaaag gccatggagt caaaaattca gatcgaggat ctaacagaac tcgccgtgaa
      481 gactggcgaa cagttcatac agagtctttt acgactcaat gacaagaaga aaatcttcgt
      541 caacatggtg gagcacgaca ctctcgtcta ctccaagaat atcaaagata cagtctcaga
      601 agaccaaagg gctattgaga cttttcaaca aagggtaata tcgggaaacc tcctcggatt
      661 ccattgccca gctatctgtc acttcatcaa aaggacagta gaaaaggaag gtggcaccta
      721 caaatgccat cattgcgata aaggaaaggc tatcgttcaa gatgcccctg ccgacagtgg
      781 tcccaaagat ggacccccac ccacgaggag catcgtggaa aaagaagacg ttccaaccac
      841 gtcttcaaag caagtggatt gatgtgatat ctccactgac gtaagggatg acgcacaatc
      901 ccactatcct tcgcaagacc cttcctctat ataaggaagt tcatttcatt tggagaggac
      961 tccggtattt ttacaacaat taccacaaca aaacaaacaa caaacaacat tacaatttac
     1021 tattctagtc gaaatggaac gagctataca aggaaacgac gctagggaac aagctaacag
     1081 tgaacgttgg gatggaggat caggaggtac cacttctccc ttcaaacttc ctgacgaaag
     1141 tccgagttgg actgagtggc ggctacataa cgatgagact aattcgaatc aagataatcc
     1201 ccttggtttc aaggaaagct ggggtttcgg gaaagttgta tttaagagat atctcagata
     1261 cgacaggacg gaagcttcac tgcacagagt ccttggatct tggacgggag attcggttaa
     1321 ctatgcagca tctcgatttt tcggtttcga ccagatcgga tgtacctata gtattcggtt
     1381 tcgaggagtt agtatcaccg tttctggagg ctctcgaact cttcagcatc tctgtgagat
     1441 ggcaattcgg tctaagcaag aactgctaca gcttgcccca atcgaagtgg aaagtaatgt
     1501 atcaagagga tgccctgaag gtactgaaac cttcgaaaaa gaaagcgagt gagcttggaa
     1561 tggatcttcg atcccgatcg ttcaaacatt tggcaataaa gtttcttaag attgaatcct
     1621 gttgccggtc ttgcgacgat tatcatataa tttctgttga attacgttaa gcatgtaata
     1681 attaacatgt aatgcatgac gttatttatg agatgggttt ttatgattag agtcccgcaa
     1741 ttatacattt aatacgcgat agaaaacaaa atatagcgcg caaactagga taaattatcg
     1801 cgcgcggtgt catctatgtt actagatcgg gaattgccaa gctaattctt gaagacgaaa
     1861 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     1921 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     1981 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatggga
     2041 ccgactcgcg cttgagacga agctttgaca ggatatattg gcgggtaaac ctaagagaaa
     2101 agagcgttta ttagaataat cggatattta aaagggcgtg aaaaggttta tccgttcgtc
     2161 catttgtatg tgcatgccaa ccacagggtt cccctcggga tcaaagtact ttgatccaac
     2221 ccctccgctg ctatagtgca gtcggcttct gacgttcagt gcagccgtca tctgaaaacg
     2281 acatgtcgca caagtcctaa gttacgcgac aggctgccgc cctgcccttt tcctggcgtt
     2341 ttcttgtcgc gtgttttagt cgcataaagt agaatacttg cgactagaac cggagacatt
     2401 acgccatgaa caagagcgcc gccgctggcc tgctgggcta tgcccgcgtc agcaccgacg
     2461 accaggactt gaccaaccaa cgggccgaac tgcacgcggc cggctgcacc aagctgtttt
     2521 ccgagaagat caccggcacc aggcgcgacc gcccggagct ggccaggatg cttgaccacc
     2581 tacgccctgg cgacgttgtg acagtgacca ggctagaccg cctggcccgc agcacccgcg
     2641 acctactgga cattgccgag cgcatccagg aggccggcgc gggcctgcgt agcctggcag
     2701 agccgtgggc cgacaccacc acgccggccg gccgcatggt gttgaccgtg ttcgccggca
     2761 ttgccgagtt cgagcgttcc ctaatcatcg accgcacccg gagcgggcgc gaggccgcca
     2821 aggcccgagg cgtgaagttt ggcccccgcc ctaccctcac cccggcacag atcgcgcacg
     2881 cccgcgagct gatcgaccag gaaggccgca ccgtgaaaga ggcggctgca ctgcttggcg
     2941 tgcatcgctc gaccctgtac cgcgcacttg agcgcagcga ggaagtgacg cccaccgagg
     3001 ccaggcggcg cggtgccttc cgtgaggacg cattgaccga ggccgacgcc ctggcggccg
     3061 ccgagaatga acgccaagag gaacaagcat gaaaccgcac caggacggcc aggacgaacc
     3121 gtttttcatt accgaagaga tcgaggcgga gatgatcgcg gccgggtacg tgttcgagcc
     3181 gcccgcgcac ctctcaaccg tgcggctgca tgaaatcctg gccggtttgt ctgatgccaa
     3241 gctggcggcc tggccggcca gcttggccgc tgaagaaacc gagcgccgcc gtctaaaaag
     3301 gtgatgtgta tttgagtaaa acagcttgcg tcatgcggtc gctgcgtata tgatccgatg
     3361 agtaaataaa caaatacgca aggggaacgc atgaaggtta tcgctgtact taaccagaaa
     3421 ggcgggtcag gcaagacgac catcggaacc catctagccc gcgccctgca actcgccggg
     3481 gccgatgttc tgttagtcga ttccgatccc cagggcagtg cccgcgattg ggcggccgtg
     3541 cgggaagatc aaccgctaac cgttgtcggc atcgaccgcc cgacgattga ccgcgacgtg
     3601 aaggccatcg gccggcgcga cttcgtagtg atcgacggag cgccccaggc ggcggacttg
     3661 gctgtgtccg cgatcaaggc agccgacttc gtgctgattc cggtgcagcc aagcccttac
     3721 gacatatggg ccaccgccga cctggtggag ctggttaagc agcgcattga ggtcacggat
     3781 ggaaggctac aagcggcctt tgtcgtgtcg cgggcgatca aaggcacgcg catcggcggt
     3841 gaggttgccg aggcgctggc cgggtacgag ctgcccattc ttgagtcccg tatcacgcag
     3901 cgcgtgagct acccaggcac tgccgccgcc ggcacaaccg ttcttgaatc agaacccgag
     3961 ggcgacgctg cccgcgaggt ccaggcgctg gccgctgaaa ttaaatcaaa actcatttga
     4021 gttaatgagg taaagagaaa atgagcaaaa gcacaaacac gctaagtgcc ggccgtccga
     4081 gcgcacgcag cagcaaggct gcaacgttgg ccagcctggc agacacgcca gccatgaagc
     4141 gggtcaactt tcagttgccg gcggaggatc acaccaagct gaagatgtac gcggtacgcc
     4201 aaggcaagac cattaccgag ctgctatctg aatagatcgc gcagctacca gagtaaatga
     4261 gcaaatgaat aaatgagtag atgaatttta gcggctaaag gaggcggcat ggaaaatcaa
     4321 gaacaaccag gcaccgacgc cgtggaatgc cccatgtgtg gaggaacggg cggttggcca
     4381 ggcgtaagcg gctgggttgt ctgccggccc tgcaatggca ctggaacccc caagcccgag
     4441 gaatcggcgt gacggtcgca aaccatccgg cccggtacaa atcggcgcgg cgctgggtga
     4501 tgacctggtg gagaagttga aggccgcgca ggccgcccag cggcaacgca tcgaggcaga
     4561 agcacgcccc ggtgaatcgt ggcaagcggc cgctgatcga atccgcaaag aatcccggca
     4621 accgccggca gccggtgcgc cgtcgattag gaagccgccc aagggcgacg agcaaccaga
     4681 ttttttcgtt ccgatgctct atgacgtggg cacccgcgat agtcgcagca tcatggacgt
     4741 ggccgttttc cgtctgtcga agcgtgaccg acgagctggc gaggtgatcc gctacgagct
     4801 tccagacggg cacgtagagg tttccgcagg gccggccggc atggccagtg tgtgggatta
     4861 cgacctggta ctgatggcgg tttcccatct aaccgaatcc atgaaccgat accgggaagg
     4921 gaagggagac aagcccggcc gcgtgttccg tccacacgtt gcggacgtac tcaagttctg
     4981 ccggcgagcc gatggcggaa agcagaaaga cgacctggta gaaacctgca ttcggttaaa
     5041 caccacgcac gttgccatgc agcgtacgaa gaaggccaag aacggccgcc tggtgacggt
     5101 atccgagggt gaagccttga ttagccgcta caagatcgta aagagcgaaa ccgggcggcc
     5161 ggagtacatc gagatcgagc tagctgattg gatgtaccgc gagatcacag aaggcaagaa
     5221 cccggacgtg ctgacggttc accccgatta ctttttgatc gatcccggca tcggccgttt
     5281 tctctaccgc ctggcacgcc gcgccgcagg caaggcagaa gccagatggt tgttcaagac
     5341 gatctacgaa cgcagtggca gcgccggaga gttcaagaag ttctgtttca ccgtgcgcaa
     5401 gctgatcggg tcaaatgacc tgccggagta cgatttgaag gaggaggcgg ggcaggctgg
     5461 cccgatccta gtcatgcgct accgcaacct gatcgagggc gaagcatccg ccggttccta
     5521 atgtacggag cagatgctag ggcaaattgc cctagcaggg gaaaaaggtc gaaaaggact
     5581 ctttcctgtg gatagcacgt acattgggaa cccaaagccg tacattggga accggaaccc
     5641 gtacattggg aacccaaagc cgtacattgg gaaccggtca cacatgtaag tgactgatat
     5701 aaaagagaaa aaaggcgatt tttccgccta aaactcttta aaacttatta aaactcttaa
     5761 aacccgcctg gcctgtgcat aactgtctgg ccagcgcaca gccgaagagc tgcaaaaagc
     5821 gcctaccctt cggtcgctgc gctccctacg ccccgccgct tcgcgtcggc ctatcgcggc
     5881 cgctggccgc tcaaaaatgg ctggcctacg gccaggcaat ctaccagggc gcggacaagc
     5941 cgcgccgtcg ccactcgacc gccggcgccc acatcaaggc accctgcctc gcgcgtttcg
     6001 gtgatgacgg tgaaaacctc tgacacatgc agctcccggt gacggtcaca gcttgtctgt
     6061 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     6121 ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc
     6181 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     6241 cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg
     6301 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     6361 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     6421 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6481 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6541 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6601 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6661 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6721 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6781 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6841 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     6901 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6961 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     7021 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     7081 gaacgaaaac tcacgttaag ggattttggt catgcattct aggtgattag aaaaactcat
     7141 cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa
     7201 aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat
     7261 cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct
     7321 cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga
     7381 atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt
     7441 catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagtc
     7501 gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca
     7561 ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct
     7621 ggaatgctgt tttccctggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga
     7681 taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct
     7741 catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat
     7801 cgggcttccc atacaatcgg tagattgtcg cacctgattg cccgacatta tcgcgagccc
     7861 atttataccc atataaatca gcatccatgt tggaatttaa tcgcggcctt gagcaagacg
     7921 tttcccgttg aatatggctc ataacagaac ttattatttc cttcctcttt tctacagtat
     7981 ttaaagatac cccaagaagc taattataac aagacgaact ccaattcact gttccttgca
     8041 ttctaaaacc ttaaatacca gaaaacagct ttttcaaagt tgttttcaaa gttggcgtat
     8101 aacatagtat cgacggagcc gattttgaaa ccgcggtgat cacaggcagc aacgctctgt
     8161 catcgttaca atcaacatgc taccctccgc gagatcatcc gtgtttcaaa cccggcagct
     8221 tagttgccgt tcttccgaat agcatcggta acatgagcaa agtctgccgc cttacaacgg
     8281 ctctcccgct gacgccgtcc cggactgatg ggctgcctgt atcgagtggt gattttgtgc
     8341 cgagctgccg gtcggggagc tgttggctgg ctggtggcag gatatattgt ggtgtaaaca
     8401 taacaagctt cgtctcagtc a
//
