LOCUS       sequence-390023-        7727 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  35S promoter-driven expression of the tomato bushy stunt virus p19 
            RNA silencing suppressor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7727)
  AUTHORS   Ganesan U, Bragg JN, Deng M, Marr S, Lee MY, Qian S, Shi M, Kappel 
            J, Peters C, Lee Y, Goodin MM, Dietzgen RG, Li Z, Jackson AO
  TITLE     Construction of a Sonchus Yellow Net Virus minireplicon: a step 
            toward reverse genetic analysis of plant negative-strand RNA 
            viruses.
  JOURNAL   J Virol. 2013 Oct;87(19):10598-611. doi: 10.1128/JVI.01397-13. Epub 
            2013 Jul 24.
  PUBMED    23885070
REFERENCE   2  (bases 1 to 7727)
  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: 
            "10.1128/JVI.01397-13"; journalName: "J Virol"; date: "2013-10"; 
            volume: "87"; issue: "19"; pages: "10598-611"
COMMENT     Sequence Label: pGD-p19
FEATURES             Location/Qualifiers
     source          1..7727
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    5..29
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        179..524
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     467..487
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      1178..1430
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    1452..1476
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             2777..3406
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3835..4908
                     /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      4974..5168
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(5404..5426)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5512..5652
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5567..5586
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5667..5684)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5838..6426)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5918..5937)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6513..7307)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
ORIGIN
        1 ctggtggcag gatatattgt ggtgtaaaca aattgacgct tagacaactt aataacacat
       61 tgcggacgtt tttaatgtac tgaattaacg ccgaattaat acgcgtcatg gtggagcacg
      121 acactcttgt ctactccaaa aatatcaaag atacagtctc agaagaccaa agggcaattg
      181 agacttttca acaaagggta atatccggaa acctcctcgg attccattgc ccagctatct
      241 gtcactttat tgtgaagata gtggaaaagg aaggtggctc ctacaaatgc catcattgcg
      301 ataaaggaaa ggccatcgtt gaagatgcct ctgccgacag tggtcccaaa gatggacccc
      361 cacccacgag gagcatcgtg gaaaaagaag acgttccaac cacgtcttca aagcaagtgg
      421 attgatgtga tatctccact gacgtaaggg atgacgcaca atcccactat ccttcgcaag
      481 acccttcctc tatataagga agttcatttc atttggagag gacacgctga aatcaccagt
      541 ctctctctac aaatctatct ctggatcccc agtctctctc tacaagatct cgagccatgg
      601 aacgagctat acaaggaaac gacgctaggg aacaagctaa cagtgaacgt tgggatggag
      661 gatcaggagg taccacttct cccttcaaac ttcctgacga aagtccgagt tggactgagt
      721 ggcggctaca taacgatgag acgaattcga atcaagataa tccccttggt ttcaaggaaa
      781 gctggggttt cgggaaagtt gtatttaaga gatatctcag atacgacagg acggaagctt
      841 cactgcacag agtccttgga tcttggacgg gagattcggt taactatgca gcatctcgat
      901 ttttcggttt cgaccagatc ggatgtacct atagtattcg gtttcgagga gttagtatca
      961 ccgtttctgg agggtcgcga actcttcagc atctctgtga gatggcaatt cggtctaagc
     1021 aagaactgct acagcttgcc ccaatcgaag tggaaagtaa tgtatcaaga ggatgccctg
     1081 aaggtactca aaccttcgaa aaagaaagcg agtaactgca gtcgacggta ccgcgggccc
     1141 gggatccacc ggatctagat aactgatcgt cgacagagct cgttcaaaca tttggcaata
     1201 aagtttctta agattgaatc ctgttgccgg tcttgcgatg attatcatat aatttctgtt
     1261 gaattacgtt aagcatgtaa taattaacat gtaatgcatg acgttattta tgagatgggt
     1321 ttttatgatt agagtcccgc aattatacat ttaatacgcg atagaaaaca aaatatagcg
     1381 cgcaaactag gataaattat cgcgcgcggt gtcatctatg ttactagatc gggaattaaa
     1441 ctatcagtgt ttgacaggat atattggcgg gtaaacctaa gagaaaagag cgtttattag
     1501 aataatcgga tatttaaaag ggcgtgaaaa ggtttatccg ttcgtccatt tgtatgtgca
     1561 tgccaaccac agggttcccc tcgggatcaa agtactttga tccaacccct ccgctgctat
     1621 agtgcagtcg gcttctgacg ttcagtgcag ccgtcttctg aaaacgacat gtcgcacaag
     1681 tcctaagtta cgcgacaggc tgccgccctg cccttttcct ggcgttttct tgtcgcgtgt
     1741 tttagtcgca taaagtagaa tacttgcgac tagaaccgga gacattacgc catgaacaag
     1801 agcgccgccg ctggcctgct gggctatgcc cgcgtcagca ccgacgacca ggacttgacc
     1861 aaccaacggg ccgaactgca cgcggccggc tgcaccaagc tgttttccga gaagatcacc
     1921 ggcaccaggc gcgaccgccc ggagctggcc aggatgcttg accacctacg ccctggcgac
     1981 gttgtgacag tgaccaggct agaccgcctg gcccgcagca cccgcgacct actggacatt
     2041 gccgagcgca tccaggaggc cggcgcgggc ctgcgtagcc tggcagagcc gtgggccgac
     2101 accaccacgc cggccggccg catggtgttg accgtgttcg ccggcattgc cgagttcgag
     2161 cgttccctaa tcatcgaccg cacccggagc gggcgcgagg ccgccaaggc ccgaggcgtg
     2221 aagtttggcc cccgccctac cctcaccccg gcacagatcg cgcacgcccg cgagctgatc
     2281 gaccaggaag gccgcaccgt gaaagaggcg gctgcactgc ttggcgtgca tcgctcgacc
     2341 ctgtaccgcg cacttgagcg cagcgaggaa gtgacgccca ccgaggccag gcggcgcggt
     2401 gccttccgtg aggacgcatt gaccgaggcc gacgccctgg cggccgccga gaatgaacgc
     2461 caagaggaac aagcatgaaa ccgcaccagg acggccagga cgaaccgttt ttcattaccg
     2521 aagagatcga ggcggagatg atcgcggccg ggtacgtgtt cgagccgccc gcgcacgtct
     2581 caaccgtgcg gctgcatgaa atcctggccg gtttgtctga tgccaagctg gcggcctggc
     2641 cggccagctt ggccgctgaa gaaaccgagc gccgccgtct aaaaaggtga tgtgtatttg
     2701 agtaaaacag cttgcgtcat gcggtcgctg cgtatatgat gcgatgagta aataaacaaa
     2761 tacgcaaggg gaacgcatga aggttatcgc tgtacttaac cagaaaggcg ggtcaggcaa
     2821 gacgaccatc gcaacccatc tagcccgcgc cctgcaactc gccggggccg atgttctgtt
     2881 agtcgattcc gatccccagg gcagtgcccg cgattgggcg gccgtgcggg aagatcaacc
     2941 gctaaccgtt gtcggcatcg accgcccgac gattgaccgc gacgtgaagg ccatcggccg
     3001 gcgcgacttc gtagtgatcg acggagcgcc ccaggcggcg gacttggctg tgtccgcgat
     3061 caaggcagcc gacttcgtgc tgattccggt gcagccaagc ccttacgaca tatgggccac
     3121 cgccgacctg gtggagctgg ttaagcagcg cattgaggtc acggatggaa ggctacaagc
     3181 ggcctttgtc gtgtcgcggg cgatcaaagg cacgcgcatc ggcggtgagg ttgccgaggc
     3241 gctggccggg tacgagctgc ccattcttga gtcccgtatc acgcagcgcg tgagctaccc
     3301 aggcactgcc gccgccggca caaccgttct tgaatcagaa cccgagggcg acgctgcccg
     3361 cgaggtccag gcgctggccg ctgaaattaa atcaaaactc atttgagtta atgaggtaaa
     3421 gagaaaatga gcaaaagcac aaacacgcta agtgccggcc gtccgagcgc acgcagcagc
     3481 aaggctgcaa cgttggccag cctggcagac acgccagcca tgaagcgggt caactttcag
     3541 ttgccggcgg aggatcacac caagctgaag atgtacgcgg tacgccaagg caagaccatt
     3601 accgagctgc tatctgaata catcgcgcag ctaccagagt aaatgagcaa atgaataaat
     3661 gagtagatga attttagcgg ctaaaggagg cggcatggaa aatcaagaac aaccaggcac
     3721 cgacgccgtg gaatgcccca tgtgtggagg aacgggcggt tggccaggcg taagcggctg
     3781 ggttgcctgc cggccctgca atggcactgg aacccccaag cccgaggaat cggcgtgagc
     3841 ggtcgcaaac catccggccc ggtacaaatc ggcgcggcgc tgggtgatga cctggtggag
     3901 aagttgaagg ccgcgcaggc cgcccagcgg caacgcatcg aggcagaagc acgccccggt
     3961 gaatcgtggc aagcggccgc tgatcgaatc cgcaaagaat cccggcaacc gccggcagcc
     4021 ggtgcgccgt cgattaggaa gccgcccaag ggcgacgagc aaccagattt tttcgttccg
     4081 atgctctatg acgtgggcac ccgcgatagt cgcagcatca tggacgtggc cgttttccgt
     4141 ctgtcgaagc gtgaccgacg agctggcgag gtgatccgct acgagcttcc agacgggcac
     4201 gtagaggttt ccgcagggcc ggccggcatg gccagtgtgt gggattacga cctggtactg
     4261 atggcggttt cccatctaac cgaatccatg aaccgatacc gggaagggaa gggagacaag
     4321 cccggccgcg tgttccgtcc acacgttgcg gacgtactca agttctgccg gcgagccgat
     4381 ggcggaaagc agaaagacga cctggtagaa acctgcattc ggttaaacac cacgcacgtt
     4441 gccatgcagc gtacgaagaa ggccaagaac ggccgcctgg tgacggtatc cgagggtgaa
     4501 gccttgatta gccgctacaa gatcgtaaag agcgaaaccg ggcggccgga gtacatcgag
     4561 atcgagctag ctgattggat gtaccgcgag atcacagaag gcaagaaccc ggacgtgctg
     4621 acggttcacc ccgattactt tttgatcgat cccggcatcg gccgttttct ctaccgcctg
     4681 gcacgccgcg ccgcaggcaa ggcagaagcc agatggttgt tcaagacgat ctacgaacgc
     4741 agtggcagcg ccggagagtt caagaagttc tgtttcaccg tgcgcaagct gatcgggtca
     4801 aatgacctgc cggagtacga tttgaaggag gaggcggggc aggctggccc gatcctagtc
     4861 atgcgctacc gcaacctgat cgagggcgaa gcatccgccg gttcctaatg tacggagcag
     4921 atgctagggc aaattgccct agcaggggaa aaaggtcgaa aaggtctctt tcctgtggat
     4981 agcacgtaca ttgggaaccc aaagccgtac attgggaacc ggaacccgta cattgggaac
     5041 ccaaagccgt acattgggaa ccggtcacac atgtaagtga ctgatataaa agagaaaaaa
     5101 ggcgattttt ccgcctaaaa ctctttaaaa cttattaaaa ctcttaaaac ccgcctggcc
     5161 tgtgcataac tgtctggcca gcgcacagcc gaagagctgc aaaaagcgcc tacccttcgg
     5221 tcgctgcgct ccctacgccc cgccgcttcg cgtcggccta tcgcggccgc tggccgctca
     5281 aaaatggctg gcctacggcc aggcaatcta ccagggcgcg gacaagccgc gccgtcgcca
     5341 ctcgaccgcc ggcgcccaca tcaaggcacc ctgcctcgcg cgtttcggtg atgacggtga
     5401 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     5461 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat
     5521 gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag
     5581 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     5641 taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5701 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5761 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5821 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5881 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5941 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6001 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6061 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6121 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6181 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6241 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     6301 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6361 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6421 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6481 cgttaaggga ttttggtcat gcattctagg tactaaaaca attcatccag taaaatataa
     6541 tattttattt tctcccaatc aggcttgatc cccagtaagt caaaaaatag ctcgacatac
     6601 tgttcttccc cgatatcctc cctgatcgac cggacgcaga aggcaatgtc ataccacttg
     6661 tccgccctgc cgcttctccc aagatcaata aagccactta ctttgccatc tttcacaaag
     6721 atgttgctgt ctcccaggtc gccgtgggaa aagacaagtt cctcttcggg cttttccgtc
     6781 tttaaaaaat catacagctc gcgcggatct ttaaatggag tgtcttcttc ccagttttcg
     6841 caatccacat cggccagatc gttattcagt aagtaatcca attcggctaa gcggctgtct
     6901 aagctattcg tatagggaca atccgatatg tcgatggagt gaaagagcct gatgcactcc
     6961 gcatacagct cgataatctt ttcagggctt tgttcatctt catactcttc cgagcaaagg
     7021 acgccatcgg cctcactcat gagcagattg ctccagccat catgccgttc aaagtgcagg
     7081 acctttggaa caggcagctt tccttccagc catagcatca tgtccttttc ccgttccaca
     7141 tcataggtgg tccctttata ccggctgtcc gtcattttta aatataggtt ttcattttct
     7201 cccaccagct tatatacctt agcaggagac attccttccg tatcttttac gcagcggtat
     7261 ttttcgatca gttttttcaa ttccggtgat attctcattt tagccattta ttatttcctt
     7321 cctcttttct acagtattta aagatacccc aagaagctaa ttataacaag acgaactcca
     7381 attcactgtt ccttgcattc taaaacctta aataccagaa aacagctttt tcaaagttgt
     7441 tttcaaagtt ggcgtataac atagtatcga cggagccgat tttgaaaccg cggtgatcac
     7501 aggcagcaac gctctgtcat cgttacaatc aacatgctac cctccgcgag atcatccgtg
     7561 tttcaaaccc ggcagcttag ttgccgttct tccgaatagc atcggtaaca tgagcaaagt
     7621 ctgccgcctt acaacggctc tcccgctgac gccgtcccgg actgatgggc tgcctgtatc
     7681 gagtggtgat tttgtgccga gctgccggtc ggggagctgt tggctgg
//
