LOCUS       sequence-390033-        7707 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  35S promoter-driven expression of the barley stripe mosaic virus 
            gamma-b RNA silencing suppressor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7707)
  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 7707)
  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-gamma-b
FEATURES             Location/Qualifiers
     source          1..7707
                     /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      1158..1410
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    1432..1456
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             2757..3386
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3815..4888
                     /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      4954..5148
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(5384..5406)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5492..5632
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5547..5566
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5647..5664)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5818..6406)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5898..5917)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6493..7287)
                     /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 aagatctcga gatgatggct actttctctt gtgtgtgttg tggtacctta
      601 actacaagta cttactgtgg taagagatgt gagcgaaagc atgtatattc tgaaacaaga
      661 aataagagat tggaacttta caagaagtat ctattggaac cgcaaaaatg cgccctgaat
      721 ggaatcgttg gacacagttg tggaatgcca tgctccattg cggaagaggc ttgtgatcaa
      781 ctgccaatcg tgagtaggtt ctgtggccaa aagcatgcgg atctgtatga ttcacttctg
      841 aaacgttctg aacaggagtt acttcttgaa tttctccaga agaagatgca ggagctgaaa
      901 ctttctcata tcgtaaaaat ggctaagctt gaaagtgagg ttaacgcaat acgtaagtcc
      961 gtagcttctt cttttgaaga ttctgttgga tgtgatgatt cttcttccgt ttctaagtaa
     1021 aaaaaaaaaa aaaatgtttg atcagatcat tcaaatctga tggtgcccat caaccatatg
     1081 atgggagtgt ttgcactgca gtcgacggta ccgcgggccc gggatccacc ggatctagat
     1141 aactgatcgt cgacagagct cgttcaaaca tttggcaata aagtttctta agattgaatc
     1201 ctgttgccgg tcttgcgatg attatcatat aatttctgtt gaattacgtt aagcatgtaa
     1261 taattaacat gtaatgcatg acgttattta tgagatgggt ttttatgatt agagtcccgc
     1321 aattatacat ttaatacgcg atagaaaaca aaatatagcg cgcaaactag gataaattat
     1381 cgcgcgcggt gtcatctatg ttactagatc gggaattaaa ctatcagtgt ttgacaggat
     1441 atattggcgg gtaaacctaa gagaaaagag cgtttattag aataatcgga tatttaaaag
     1501 ggcgtgaaaa ggtttatccg ttcgtccatt tgtatgtgca tgccaaccac agggttcccc
     1561 tcgggatcaa agtactttga tccaacccct ccgctgctat agtgcagtcg gcttctgacg
     1621 ttcagtgcag ccgtcttctg aaaacgacat gtcgcacaag tcctaagtta cgcgacaggc
     1681 tgccgccctg cccttttcct ggcgttttct tgtcgcgtgt tttagtcgca taaagtagaa
     1741 tacttgcgac tagaaccgga gacattacgc catgaacaag agcgccgccg ctggcctgct
     1801 gggctatgcc cgcgtcagca ccgacgacca ggacttgacc aaccaacggg ccgaactgca
     1861 cgcggccggc tgcaccaagc tgttttccga gaagatcacc ggcaccaggc gcgaccgccc
     1921 ggagctggcc aggatgcttg accacctacg ccctggcgac gttgtgacag tgaccaggct
     1981 agaccgcctg gcccgcagca cccgcgacct actggacatt gccgagcgca tccaggaggc
     2041 cggcgcgggc ctgcgtagcc tggcagagcc gtgggccgac accaccacgc cggccggccg
     2101 catggtgttg accgtgttcg ccggcattgc cgagttcgag cgttccctaa tcatcgaccg
     2161 cacccggagc gggcgcgagg ccgccaaggc ccgaggcgtg aagtttggcc cccgccctac
     2221 cctcaccccg gcacagatcg cgcacgcccg cgagctgatc gaccaggaag gccgcaccgt
     2281 gaaagaggcg gctgcactgc ttggcgtgca tcgctcgacc ctgtaccgcg cacttgagcg
     2341 cagcgaggaa gtgacgccca ccgaggccag gcggcgcggt gccttccgtg aggacgcatt
     2401 gaccgaggcc gacgccctgg cggccgccga gaatgaacgc caagaggaac aagcatgaaa
     2461 ccgcaccagg acggccagga cgaaccgttt ttcattaccg aagagatcga ggcggagatg
     2521 atcgcggccg ggtacgtgtt cgagccgccc gcgcacgtct caaccgtgcg gctgcatgaa
     2581 atcctggccg gtttgtctga tgccaagctg gcggcctggc cggccagctt ggccgctgaa
     2641 gaaaccgagc gccgccgtct aaaaaggtga tgtgtatttg agtaaaacag cttgcgtcat
     2701 gcggtcgctg cgtatatgat gcgatgagta aataaacaaa tacgcaaggg gaacgcatga
     2761 aggttatcgc tgtacttaac cagaaaggcg ggtcaggcaa gacgaccatc gcaacccatc
     2821 tagcccgcgc cctgcaactc gccggggccg atgttctgtt agtcgattcc gatccccagg
     2881 gcagtgcccg cgattgggcg gccgtgcggg aagatcaacc gctaaccgtt gtcggcatcg
     2941 accgcccgac gattgaccgc gacgtgaagg ccatcggccg gcgcgacttc gtagtgatcg
     3001 acggagcgcc ccaggcggcg gacttggctg tgtccgcgat caaggcagcc gacttcgtgc
     3061 tgattccggt gcagccaagc ccttacgaca tatgggccac cgccgacctg gtggagctgg
     3121 ttaagcagcg cattgaggtc acggatggaa ggctacaagc ggcctttgtc gtgtcgcggg
     3181 cgatcaaagg cacgcgcatc ggcggtgagg ttgccgaggc gctggccggg tacgagctgc
     3241 ccattcttga gtcccgtatc acgcagcgcg tgagctaccc aggcactgcc gccgccggca
     3301 caaccgttct tgaatcagaa cccgagggcg acgctgcccg cgaggtccag gcgctggccg
     3361 ctgaaattaa atcaaaactc atttgagtta atgaggtaaa gagaaaatga gcaaaagcac
     3421 aaacacgcta agtgccggcc gtccgagcgc acgcagcagc aaggctgcaa cgttggccag
     3481 cctggcagac acgccagcca tgaagcgggt caactttcag ttgccggcgg aggatcacac
     3541 caagctgaag atgtacgcgg tacgccaagg caagaccatt accgagctgc tatctgaata
     3601 catcgcgcag ctaccagagt aaatgagcaa atgaataaat gagtagatga attttagcgg
     3661 ctaaaggagg cggcatggaa aatcaagaac aaccaggcac cgacgccgtg gaatgcccca
     3721 tgtgtggagg aacgggcggt tggccaggcg taagcggctg ggttgcctgc cggccctgca
     3781 atggcactgg aacccccaag cccgaggaat cggcgtgagc ggtcgcaaac catccggccc
     3841 ggtacaaatc ggcgcggcgc tgggtgatga cctggtggag aagttgaagg ccgcgcaggc
     3901 cgcccagcgg caacgcatcg aggcagaagc acgccccggt gaatcgtggc aagcggccgc
     3961 tgatcgaatc cgcaaagaat cccggcaacc gccggcagcc ggtgcgccgt cgattaggaa
     4021 gccgcccaag ggcgacgagc aaccagattt tttcgttccg atgctctatg acgtgggcac
     4081 ccgcgatagt cgcagcatca tggacgtggc cgttttccgt ctgtcgaagc gtgaccgacg
     4141 agctggcgag gtgatccgct acgagcttcc agacgggcac gtagaggttt ccgcagggcc
     4201 ggccggcatg gccagtgtgt gggattacga cctggtactg atggcggttt cccatctaac
     4261 cgaatccatg aaccgatacc gggaagggaa gggagacaag cccggccgcg tgttccgtcc
     4321 acacgttgcg gacgtactca agttctgccg gcgagccgat ggcggaaagc agaaagacga
     4381 cctggtagaa acctgcattc ggttaaacac cacgcacgtt gccatgcagc gtacgaagaa
     4441 ggccaagaac ggccgcctgg tgacggtatc cgagggtgaa gccttgatta gccgctacaa
     4501 gatcgtaaag agcgaaaccg ggcggccgga gtacatcgag atcgagctag ctgattggat
     4561 gtaccgcgag atcacagaag gcaagaaccc ggacgtgctg acggttcacc ccgattactt
     4621 tttgatcgat cccggcatcg gccgttttct ctaccgcctg gcacgccgcg ccgcaggcaa
     4681 ggcagaagcc agatggttgt tcaagacgat ctacgaacgc agtggcagcg ccggagagtt
     4741 caagaagttc tgtttcaccg tgcgcaagct gatcgggtca aatgacctgc cggagtacga
     4801 tttgaaggag gaggcggggc aggctggccc gatcctagtc atgcgctacc gcaacctgat
     4861 cgagggcgaa gcatccgccg gttcctaatg tacggagcag atgctagggc aaattgccct
     4921 agcaggggaa aaaggtcgaa aaggtctctt tcctgtggat agcacgtaca ttgggaaccc
     4981 aaagccgtac attgggaacc ggaacccgta cattgggaac ccaaagccgt acattgggaa
     5041 ccggtcacac atgtaagtga ctgatataaa agagaaaaaa ggcgattttt ccgcctaaaa
     5101 ctctttaaaa cttattaaaa ctcttaaaac ccgcctggcc tgtgcataac tgtctggcca
     5161 gcgcacagcc gaagagctgc aaaaagcgcc tacccttcgg tcgctgcgct ccctacgccc
     5221 cgccgcttcg cgtcggccta tcgcggccgc tggccgctca aaaatggctg gcctacggcc
     5281 aggcaatcta ccagggcgcg gacaagccgc gccgtcgcca ctcgaccgcc ggcgcccaca
     5341 tcaaggcacc ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     5401 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     5461 gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata
     5521 gcggagtgta tactggctta actatgcggc atcagagcag attgtactga gagtgcacca
     5581 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc
     5641 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5701 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5761 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5821 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5881 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5941 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6001 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6061 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6121 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6181 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6241 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6301 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6361 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6421 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6481 gcattctagg tactaaaaca attcatccag taaaatataa tattttattt tctcccaatc
     6541 aggcttgatc cccagtaagt caaaaaatag ctcgacatac tgttcttccc cgatatcctc
     6601 cctgatcgac cggacgcaga aggcaatgtc ataccacttg tccgccctgc cgcttctccc
     6661 aagatcaata aagccactta ctttgccatc tttcacaaag atgttgctgt ctcccaggtc
     6721 gccgtgggaa aagacaagtt cctcttcggg cttttccgtc tttaaaaaat catacagctc
     6781 gcgcggatct ttaaatggag tgtcttcttc ccagttttcg caatccacat cggccagatc
     6841 gttattcagt aagtaatcca attcggctaa gcggctgtct aagctattcg tatagggaca
     6901 atccgatatg tcgatggagt gaaagagcct gatgcactcc gcatacagct cgataatctt
     6961 ttcagggctt tgttcatctt catactcttc cgagcaaagg acgccatcgg cctcactcat
     7021 gagcagattg ctccagccat catgccgttc aaagtgcagg acctttggaa caggcagctt
     7081 tccttccagc catagcatca tgtccttttc ccgttccaca tcataggtgg tccctttata
     7141 ccggctgtcc gtcattttta aatataggtt ttcattttct cccaccagct tatatacctt
     7201 agcaggagac attccttccg tatcttttac gcagcggtat ttttcgatca gttttttcaa
     7261 ttccggtgat attctcattt tagccattta ttatttcctt cctcttttct acagtattta
     7321 aagatacccc aagaagctaa ttataacaag acgaactcca attcactgtt ccttgcattc
     7381 taaaacctta aataccagaa aacagctttt tcaaagttgt tttcaaagtt ggcgtataac
     7441 atagtatcga cggagccgat tttgaaaccg cggtgatcac aggcagcaac gctctgtcat
     7501 cgttacaatc aacatgctac cctccgcgag atcatccgtg tttcaaaccc ggcagcttag
     7561 ttgccgttct tccgaatagc atcggtaaca tgagcaaagt ctgccgcctt acaacggctc
     7621 tcccgctgac gccgtcccgg actgatgggc tgcctgtatc gagtggtgat tttgtgccga
     7681 gctgccggtc ggggagctgt tggctgg
//
