LOCUS       sequence-176326-        5664 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Rotavirus SA11 VP3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5664)
  AUTHORS   Kanai Y, Komoto S, Kawagishi T, Nouda R, Nagasawa N, Onishi M, 
            Matsuura Y, Taniguchi K, Kobayashi T
  TITLE     Entirely plasmid-based reverse genetics system for rotaviruses.
  JOURNAL   Proc Natl Acad Sci U S A. 2017 Jan 30. pii: 201618424. doi: 
            10.1073/pnas.1618424114.
  PUBMED    28137864
REFERENCE   2  (bases 1 to 5664)
  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: "Proc Natl 
            Acad Sci U S A. 2017 Jan 30. pii: 201618424. doi: 
            10.1073/pnas.1618424114."
COMMENT     Sequence Label: pT7-VP3SA11
FEATURES             Location/Qualifiers
     source          1..5664
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(167..755)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(247..266)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(926..1717,1718..1786))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1549..1568
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1787..1891)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1959..1977
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2015..2037)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2137..2156
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2366..2385
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2675..2693
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2675..2692
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      complement(2761..2808)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2804..2822
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     promoter        complement(5558..5576)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(join(5660..5664,1..13))
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
       61 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
      121 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
      181 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
      241 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
      301 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
      361 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
      421 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
      481 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
      541 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
      601 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
      661 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
      721 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
      781 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
      841 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
      901 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
      961 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     1021 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     1081 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     1141 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     1201 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     1261 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     1321 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     1381 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     1441 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     1501 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     1561 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     1621 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     1681 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     1741 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     1801 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     1861 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     1921 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     1981 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     2041 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     2101 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     2161 gagtgcacca ttcgacgctc tcccttatgc gactcctgca ttaagaagca gcccagtagt
     2221 aggttgaggc cgttgagcac cgccgccgca aggaatggtg caaggagatg gcgcccaaca
     2281 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
     2341 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
     2401 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatctgg ctagcgatga
     2461 ccctgctgat tggttcgctg accatttccg ggtgcgggac ggcgttacca gaaactcaga
     2521 aggttcgtcc aaccaaaccg actctgacgg cagtttacga gagagatgat agggtctgct
     2581 tcagtaagcc agatgctaca caattaggct tgtacatatt gtcgttagaa cgcggctaca
     2641 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaatacaag
     2701 cttgcatgcc tgcaggtcga ctctagagga tctcgatccg gatatagttc ctcctttcag
     2761 caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctagtt attgctcagc
     2821 ggtggcagca gccaactcag cttcctttcg ggctttgtta gcagccggat cctttttttt
     2881 gagctctccc ttagccatcc gagtggacga cgtcctcctt cggatgccca ggtcggaccg
     2941 cgaggaggtg gagatgccat gccgacccgg tcacatcatg actagtgtgt taagttttta
     3001 gctcactcaa ccatatcaaa cactgtatca cccttaatct ttcttaaata aatggattta
     3061 atttgtaatt tagtagtctc tatccttact gttctcatgt tattgaagct taatgttatg
     3121 tgcggaataa acattctgga ttggccaaaa gttatttgat ttttataaaa tacgttatag
     3181 attttaaagt atgtagataa tactccacta acatttgcta cgtagattcc atctgataac
     3241 atatatgtat aactggtcgt aatatccatt ttaaataatt cattcttcat tatctttagt
     3301 atgtcaatca cgtctggcac catgttatca gaaatatgta atactccgaa ggttaaatgc
     3361 ggtattttaa catcatattt atatggtata tttctaaatt gtattccgat aaagtaattc
     3421 ggatcatcag cgtatgttat actatatact gtattaatat aattctctat ttcgtttgaa
     3481 tatctcaata cagttaagtc atcctgcaat gtagcaaatt cttttgctga tctacatgca
     3541 taaattaatt ctattggtgc gtgttcataa tactttcctc cttctatatc aactttatct
     3601 atagaatgta aataaatcca gcgtttaaga tcaaatgaat aattatatct ataataaatt
     3661 aaagcattat atacatgacc tgatactgaa ttctctatat tagtaccaat tagattaaaa
     3721 ttattattgg acaaatctct aaatatataa cctgaatatg aagtagcaga ttccgaaaac
     3781 ctaatttggt gtgatcttct cgatatactg gtgtgattag caaattgatc caatttataa
     3841 tacttatctg taccacttaa aatgaataaa tgattattcc ctgttggttt tgatgctaac
     3901 gatatagaca aaccaaagag tcctaaacaa ccgtcgtatg aagtaattat agcttctttg
     3961 gtatcaaagt cggttggaag aaaggtccaa tcatagatat ttttgaaccc aacttttggt
     4021 tcatccttaa acgtattatt tattctaaca gttattagag attgtttctg attattaatt
     4081 aatttaatta cttctgatct attattaaaa tcatttgata atgcatataa cgctacaata
     4141 taactatcat tcagtacttt tactttaaat ggttgttgaa taaacacatt ttcagttatt
     4201 atattgttta taaatgaata taacacgcct ttaggaatga acctcctaat gttagttaaa
     4261 tcccaagtat ctagtaataa ataaaattct gatcttattt ccgtagttgg gtggtgcagt
     4321 aatttagctg aaattggcag ttccaaatcc atagctgtca tcttaacaca acacaccttc
     4381 gctttacccg ttcgtaaata ttcataagct atgtccaaat tattaatagt ttgttcttct
     4441 actgtctttc tccatttttt ccaatcagca ttgcccctat cggttcttat atcaatatac
     4501 aatattgagt tatccttcag tttcttcaca tcatttataa aaaataattc tttgtgatgg
     4561 attgatgaat atggtgtgtc caatggatcc caagtttcaa agaaaaaatc ggaataatgc
     4621 ctaacgtcat atatccaata agatggagca gatccgatca tatatataat tggtttactt
     4681 ttatataaat tatagtaata ataaattgct gtaagaacat tcagcatatt tttatactga
     4741 ccaatatcaa actgagatat ccttttacct aaccatcgtt cttgttttaa ttttatagta
     4801 tcttttacta aatacgcacc atttgaaaat tgttttatag ttggataatt taaaaatgta
     4861 aacatagatt cattacgcgg accaacgttc actacattct tatgttcttt caataatgat
     4921 gggaataact tatccctaaa tactctgtta gataatctag ctataggcaa cgatgtagtc
     4981 attctttcct ttatcttttt taacataaaa ttttggaagt ccaacgcttc tgaataaatg
     5041 tattcaatta ttagatcatc atctgttgca gcgttttgac atttaaatga aaagttataa
     5101 tgtcctctag tttcacttcc attataacca acatatgtta atttccatcc aggtagtaaa
     5161 tattcttcaa tatttgtttt tgttatccaa tgttggtctg ttctaatttc attattggta
     5221 taatatgaat aatcatgtaa atatattata tcaagttcat aatcataagt gaaattacac
     5281 cttattaatg tgaataattc ttcaagtgat tgtaaagcaa ttgcagctat tcctgactta
     5341 tttaatattt ctaatgtttt aatgctataa tttaagtata aaatattatg ggtcgtaaga
     5401 ttagagatta aaaaagcgtt ttcataacta tcttttgtat catcatgaac gtagacttga
     5461 gtgtctgcat acacttgagc cacactgtgt cttaaagcta gtactttcat gtttacacca
     5521 tcagaggtaa aacacactac tggtactgct ttaatagcct atagtgagtc gtattaaccg
     5581 ggatcgatat ccccctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5641 ttgggcgctc ttccgcttcc tcgc
//
