LOCUS       sequence-390012-        9855 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  35S promoter-driven expression of the tobacco etch virus P1/HC-Pro 
            RNA silencing suppressor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9855)
  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 9855)
  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-P1/HC-Pro
FEATURES             Location/Qualifiers
     source          1..9855
                     /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      3306..3558
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    3580..3604
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             4905..5534
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             5963..7036
                     /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      7102..7296
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(7532..7554)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    7640..7780
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     7695..7714
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7795..7812)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7966..8554)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8046..8065)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8641..9435)
                     /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 cgagaattct
      601 caacacaaca tatacaaaac aaacgaatct caagcaatca agcattctac ttctattgca
      661 gcaatttaaa tcatttcttt taaagcaaaa gcaattttct gaaaattttc accatttacg
      721 aacgatagcc atggcactca tctttggcac agtcaacgct aacatcctga aggaagtgtt
      781 cggtggagct cgtatggctt gcgttaccag cgcacatatg gctggagcga atggaagcat
      841 tttgaagaag gcagaagaga cctctcgtgc aatcatgcac aaaccagtga tcttcggaga
      901 agactacatt accgaggcag acttgcctta cacaccactc catttagagg tcgatgctga
      961 aatggagcgg atgtattatc ttggtcgtcg cgcgctcacc catggcaaga gacgcaaagt
     1021 ttctgtgaat aacaagagga acaggagaag gaaagtggcc aaaacgtacg tggggcgtga
     1081 ttccattgtt gagaagattg tagtgcccca caccgagaga aaggttgata ccacagcagc
     1141 agtggaagac atttgcaatg aagctaccac tcaacttgtg cataatagta tgccaaagcg
     1201 taagaagcag aaaaacttct tgcccgccac ttcactaagt aacgtgtatg cccaaacttg
     1261 gagcatagtg cgcaaacgcc atatgcaggt ggagatcatt agcaagaaga gcgtccgagc
     1321 gagggtcaag agatttgagg gctcggtgca attgttcgca agtgtgcgtc acatgtatgg
     1381 cgagaggaaa agggtggact tacgtattga caactggcag caagagacac ttctagacct
     1441 tgctaaaaga tttaagaatg agagagtgga tcaatcgaag ctcacttttg gttcaagtgg
     1501 cctagttttg aggcaaggct cgtacggacc tgcgcattgg tatcgacatg gtatgttcat
     1561 tgtacgcggt cggtcggatg ggatgttggt ggatgctcgt gcgaaggtaa cgttcgctgt
     1621 ttgtcactca atgacacatt atagcgacaa atcaatctct gaggcattct tcataccata
     1681 ctctaagaaa ttcttggagt tgagaccaga tggaatctcc catgagtgta caagaggagt
     1741 atcagttgag cggtgcggtg aggtggctgc aatcctgaca caagcacttt caccgtgtgg
     1801 taagatcaca tgcaaacgtt gcatggttga aacacctgac attgttgagg gtgagtcggg
     1861 agacagtgtc accaaccaag gtaagctcct agcaatgctg aaagaacagt atccagattt
     1921 cccaatggcc gagaaactac tcacaaggtt tttgcaacag aaatcactag taaatacaaa
     1981 tttgacagcc tgcgtgagcg tcaaacaact cattggtgac cgcaaacaag ctccattcac
     2041 acacgtactg gctgtcagcg aaattctgtt taaaggcaat aaactaacag gggctgatct
     2101 cgaagaggca agcacacata tgcttgaaat agcaaggttc ttgaacaatc gcactgaaaa
     2161 tatgcgcatt ggccaccttg gttctttcag aaataaaatc tcatcgaagg cccatgtgaa
     2221 taacgcactc atgtgtgata atcaacttga tcagaatggg aattttattt ggggactaag
     2281 gggtgcacac gcaaagaggt ttcttaaagg atttttcact gagattgacc caaatgaagg
     2341 atacgataag tatgttatca ggaaacatat caggggtagc agaaagctag caattggcaa
     2401 tttgataatg tcaactgact tccagacgct caggcaacaa attcaaggcg aaactattga
     2461 gcgtaaagaa attgggaatc actgcatttc aatgcggaat ggtaattacg tgtacccatg
     2521 ttgttgtgtt actcttgaag atggtaaggc tcaatattcg gatctaaagc acccaacgaa
     2581 gagacatctg gtcattggca actctggcga ttcaaagtac ctagaccttc cagttctcaa
     2641 tgaagagaaa atgtatatag ctaatgaagg ttattgctac atgaacattt tctttgctct
     2701 actagtgaat gtcaaggaag aggatgcaaa ggacttcacc aagtttataa gggacacaat
     2761 tgttccaaag cttggagcgt ggccaacaat gcaagatgtt gcaactgcat gctacttact
     2821 ttccattctt tacccagatg tcctgagtgc tgaactaccc agaattttgg ttgatcatga
     2881 caacaaaaca atgcatgttt tggattcgta tgggtctaga acgacaggat accacatgtt
     2941 gaaaatgaac acaacatccc agctaattga attcgttcat tcaggtttgg aatccgaaat
     3001 gaaaacttac aatgttggag ggatgaaccg agatatggtc acacaaggtg caattgagat
     3061 gttgatcaag tctatataca aaccacatct catgaagcag ttacttgagg aggagccata
     3121 cataattgtc ctggcaatag tctccccttc aattttaatt gccatgtaca actctggaac
     3181 ttttgagcag gcgttacaaa tgtggttgcc aaatacaatg aggttagcta acctcgctgc
     3241 catcttgtca gccttagcgc aaaagttggg gatccaccgg atctagataa ctgatcgtcg
     3301 acagagctcg ttcaaacatt tggcaataaa gtttcttaag attgaatcct gttgccggtc
     3361 ttgcgatgat tatcatataa tttctgttga attacgttaa gcatgtaata attaacatgt
     3421 aatgcatgac gttatttatg agatgggttt ttatgattag agtcccgcaa ttatacattt
     3481 aatacgcgat agaaaacaaa atatagcgcg caaactagga taaattatcg cgcgcggtgt
     3541 catctatgtt actagatcgg gaattaaact atcagtgttt gacaggatat attggcgggt
     3601 aaacctaaga gaaaagagcg tttattagaa taatcggata tttaaaaggg cgtgaaaagg
     3661 tttatccgtt cgtccatttg tatgtgcatg ccaaccacag ggttcccctc gggatcaaag
     3721 tactttgatc caacccctcc gctgctatag tgcagtcggc ttctgacgtt cagtgcagcc
     3781 gtcttctgaa aacgacatgt cgcacaagtc ctaagttacg cgacaggctg ccgccctgcc
     3841 cttttcctgg cgttttcttg tcgcgtgttt tagtcgcata aagtagaata cttgcgacta
     3901 gaaccggaga cattacgcca tgaacaagag cgccgccgct ggcctgctgg gctatgcccg
     3961 cgtcagcacc gacgaccagg acttgaccaa ccaacgggcc gaactgcacg cggccggctg
     4021 caccaagctg ttttccgaga agatcaccgg caccaggcgc gaccgcccgg agctggccag
     4081 gatgcttgac cacctacgcc ctggcgacgt tgtgacagtg accaggctag accgcctggc
     4141 ccgcagcacc cgcgacctac tggacattgc cgagcgcatc caggaggccg gcgcgggcct
     4201 gcgtagcctg gcagagccgt gggccgacac caccacgccg gccggccgca tggtgttgac
     4261 cgtgttcgcc ggcattgccg agttcgagcg ttccctaatc atcgaccgca cccggagcgg
     4321 gcgcgaggcc gccaaggccc gaggcgtgaa gtttggcccc cgccctaccc tcaccccggc
     4381 acagatcgcg cacgcccgcg agctgatcga ccaggaaggc cgcaccgtga aagaggcggc
     4441 tgcactgctt ggcgtgcatc gctcgaccct gtaccgcgca cttgagcgca gcgaggaagt
     4501 gacgcccacc gaggccaggc ggcgcggtgc cttccgtgag gacgcattga ccgaggccga
     4561 cgccctggcg gccgccgaga atgaacgcca agaggaacaa gcatgaaacc gcaccaggac
     4621 ggccaggacg aaccgttttt cattaccgaa gagatcgagg cggagatgat cgcggccggg
     4681 tacgtgttcg agccgcccgc gcacgtctca accgtgcggc tgcatgaaat cctggccggt
     4741 ttgtctgatg ccaagctggc ggcctggccg gccagcttgg ccgctgaaga aaccgagcgc
     4801 cgccgtctaa aaaggtgatg tgtatttgag taaaacagct tgcgtcatgc ggtcgctgcg
     4861 tatatgatgc gatgagtaaa taaacaaata cgcaagggga acgcatgaag gttatcgctg
     4921 tacttaacca gaaaggcggg tcaggcaaga cgaccatcgc aacccatcta gcccgcgccc
     4981 tgcaactcgc cggggccgat gttctgttag tcgattccga tccccagggc agtgcccgcg
     5041 attgggcggc cgtgcgggaa gatcaaccgc taaccgttgt cggcatcgac cgcccgacga
     5101 ttgaccgcga cgtgaaggcc atcggccggc gcgacttcgt agtgatcgac ggagcgcccc
     5161 aggcggcgga cttggctgtg tccgcgatca aggcagccga cttcgtgctg attccggtgc
     5221 agccaagccc ttacgacata tgggccaccg ccgacctggt ggagctggtt aagcagcgca
     5281 ttgaggtcac ggatggaagg ctacaagcgg cctttgtcgt gtcgcgggcg atcaaaggca
     5341 cgcgcatcgg cggtgaggtt gccgaggcgc tggccgggta cgagctgccc attcttgagt
     5401 cccgtatcac gcagcgcgtg agctacccag gcactgccgc cgccggcaca accgttcttg
     5461 aatcagaacc cgagggcgac gctgcccgcg aggtccaggc gctggccgct gaaattaaat
     5521 caaaactcat ttgagttaat gaggtaaaga gaaaatgagc aaaagcacaa acacgctaag
     5581 tgccggccgt ccgagcgcac gcagcagcaa ggctgcaacg ttggccagcc tggcagacac
     5641 gccagccatg aagcgggtca actttcagtt gccggcggag gatcacacca agctgaagat
     5701 gtacgcggta cgccaaggca agaccattac cgagctgcta tctgaataca tcgcgcagct
     5761 accagagtaa atgagcaaat gaataaatga gtagatgaat tttagcggct aaaggaggcg
     5821 gcatggaaaa tcaagaacaa ccaggcaccg acgccgtgga atgccccatg tgtggaggaa
     5881 cgggcggttg gccaggcgta agcggctggg ttgcctgccg gccctgcaat ggcactggaa
     5941 cccccaagcc cgaggaatcg gcgtgagcgg tcgcaaacca tccggcccgg tacaaatcgg
     6001 cgcggcgctg ggtgatgacc tggtggagaa gttgaaggcc gcgcaggccg cccagcggca
     6061 acgcatcgag gcagaagcac gccccggtga atcgtggcaa gcggccgctg atcgaatccg
     6121 caaagaatcc cggcaaccgc cggcagccgg tgcgccgtcg attaggaagc cgcccaaggg
     6181 cgacgagcaa ccagattttt tcgttccgat gctctatgac gtgggcaccc gcgatagtcg
     6241 cagcatcatg gacgtggccg ttttccgtct gtcgaagcgt gaccgacgag ctggcgaggt
     6301 gatccgctac gagcttccag acgggcacgt agaggtttcc gcagggccgg ccggcatggc
     6361 cagtgtgtgg gattacgacc tggtactgat ggcggtttcc catctaaccg aatccatgaa
     6421 ccgataccgg gaagggaagg gagacaagcc cggccgcgtg ttccgtccac acgttgcgga
     6481 cgtactcaag ttctgccggc gagccgatgg cggaaagcag aaagacgacc tggtagaaac
     6541 ctgcattcgg ttaaacacca cgcacgttgc catgcagcgt acgaagaagg ccaagaacgg
     6601 ccgcctggtg acggtatccg agggtgaagc cttgattagc cgctacaaga tcgtaaagag
     6661 cgaaaccggg cggccggagt acatcgagat cgagctagct gattggatgt accgcgagat
     6721 cacagaaggc aagaacccgg acgtgctgac ggttcacccc gattactttt tgatcgatcc
     6781 cggcatcggc cgttttctct accgcctggc acgccgcgcc gcaggcaagg cagaagccag
     6841 atggttgttc aagacgatct acgaacgcag tggcagcgcc ggagagttca agaagttctg
     6901 tttcaccgtg cgcaagctga tcgggtcaaa tgacctgccg gagtacgatt tgaaggagga
     6961 ggcggggcag gctggcccga tcctagtcat gcgctaccgc aacctgatcg agggcgaagc
     7021 atccgccggt tcctaatgta cggagcagat gctagggcaa attgccctag caggggaaaa
     7081 aggtcgaaaa ggtctctttc ctgtggatag cacgtacatt gggaacccaa agccgtacat
     7141 tgggaaccgg aacccgtaca ttgggaaccc aaagccgtac attgggaacc ggtcacacat
     7201 gtaagtgact gatataaaag agaaaaaagg cgatttttcc gcctaaaact ctttaaaact
     7261 tattaaaact cttaaaaccc gcctggcctg tgcataactg tctggccagc gcacagccga
     7321 agagctgcaa aaagcgccta cccttcggtc gctgcgctcc ctacgccccg ccgcttcgcg
     7381 tcggcctatc gcggccgctg gccgctcaaa aatggctggc ctacggccag gcaatctacc
     7441 agggcgcgga caagccgcgc cgtcgccact cgaccgccgg cgcccacatc aaggcaccct
     7501 gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     7561 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     7621 gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata
     7681 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     7741 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct
     7801 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     7861 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     7921 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     7981 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     8041 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     8101 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     8161 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     8221 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     8281 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     8341 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     8401 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     8461 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     8521 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     8581 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatgc attctaggta
     8641 ctaaaacaat tcatccagta aaatataata ttttattttc tcccaatcag gcttgatccc
     8701 cagtaagtca aaaaatagct cgacatactg ttcttccccg atatcctccc tgatcgaccg
     8761 gacgcagaag gcaatgtcat accacttgtc cgccctgccg cttctcccaa gatcaataaa
     8821 gccacttact ttgccatctt tcacaaagat gttgctgtct cccaggtcgc cgtgggaaaa
     8881 gacaagttcc tcttcgggct tttccgtctt taaaaaatca tacagctcgc gcggatcttt
     8941 aaatggagtg tcttcttccc agttttcgca atccacatcg gccagatcgt tattcagtaa
     9001 gtaatccaat tcggctaagc ggctgtctaa gctattcgta tagggacaat ccgatatgtc
     9061 gatggagtga aagagcctga tgcactccgc atacagctcg ataatctttt cagggctttg
     9121 ttcatcttca tactcttccg agcaaaggac gccatcggcc tcactcatga gcagattgct
     9181 ccagccatca tgccgttcaa agtgcaggac ctttggaaca ggcagctttc cttccagcca
     9241 tagcatcatg tccttttccc gttccacatc ataggtggtc cctttatacc ggctgtccgt
     9301 catttttaaa tataggtttt cattttctcc caccagctta tataccttag caggagacat
     9361 tccttccgta tcttttacgc agcggtattt ttcgatcagt tttttcaatt ccggtgatat
     9421 tctcatttta gccatttatt atttccttcc tcttttctac agtatttaaa gataccccaa
     9481 gaagctaatt ataacaagac gaactccaat tcactgttcc ttgcattcta aaaccttaaa
     9541 taccagaaaa cagctttttc aaagttgttt tcaaagttgg cgtataacat agtatcgacg
     9601 gagccgattt tgaaaccgcg gtgatcacag gcagcaacgc tctgtcatcg ttacaatcaa
     9661 catgctaccc tccgcgagat catccgtgtt tcaaacccgg cagcttagtt gccgttcttc
     9721 cgaatagcat cggtaacatg agcaaagtct gccgccttac aacggctctc ccgctgacgc
     9781 cgtcccggac tgatgggctg cctgtatcga gtggtgattt tgtgccgagc tgccggtcgg
     9841 ggagctgttg gctgg
//
