LOCUS       sequence-176101-        5436 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Rotavirus SA11 VP4 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5436)
  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 5436)
  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-VP4SA11
FEATURES             Location/Qualifiers
     source          1..5436
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(48..636)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(128..147)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(807..1598,1599..1667))
                     /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     1430..1449
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1668..1772)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1840..1858
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1896..1918)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2018..2037
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2247..2266
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2556..2574
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2556..2573
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      complement(2642..2689)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2685..2703
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     promoter        complement(5210..5228)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5313..5330)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
       61 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
      121 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
      181 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
      241 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
      301 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
      361 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
      421 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
      481 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
      541 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
      601 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
      661 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
      721 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
      781 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
      841 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
      901 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
      961 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     1021 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     1081 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     1141 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     1201 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     1261 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     1321 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     1381 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     1441 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     1501 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     1561 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     1621 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     1681 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     1741 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     1801 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     1861 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     1921 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     1981 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     2041 agagtgcacc attcgacgct ctcccttatg cgactcctgc attaagaagc agcccagtag
     2101 taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcaaggagat ggcgcccaac
     2161 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
     2221 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
     2281 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatctg gctagcgatg
     2341 accctgctga ttggttcgct gaccatttcc gggtgcggga cggcgttacc agaaactcag
     2401 aaggttcgtc caaccaaacc gactctgacg gcagtttacg agagagatga tagggtctgc
     2461 ttcagtaagc cagatgctac acaattaggc ttgtacatat tgtcgttaga acgcggctac
     2521 aattaataca taaccttatg tatcatacac atacgattta ggtgacacta tagaatacaa
     2581 gcttgcatgc ctgcaggtcg actctagagg atctcgatcc ggatatagtt cctcctttca
     2641 gcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctagt tattgctcag
     2701 cggtggcagc agccaactca gcttcctttc gggctttgtt agcagccgga tccttttttt
     2761 tgagctctcc cttagccatc cgagtggacg acgtcctcct tcggatgccc aggtcggacc
     2821 gcgaggaggt ggagatgcca tgccgacccg gtcacatcct ctagaaatta ctcacaacct
     2881 gcattgcata atcagttgtt caattctatt tctaattata ggattgtcct gattaatgaa
     2941 ttcacgtaaa actcttgggt cagatcttaa aagattaaat gcttgttgct tagtaatgcc
     3001 gtaattatcg ttcaaatttt taagtgtttt aaaatcaatt atagcggata ttactggaga
     3061 atctgtaact aagtcagcga atttttgtac atcaaatggt atttcctcaa atgtatccac
     3121 tttataagca aaaaatttcc catcaattcc agcttcaaac acatcatcgt tgtttataac
     3181 gcggtaagcc ctatttggta tgaatttttc cgatgcttca gtaacaatgt caggtattgt
     3241 gtttggtgat atttgagtcg atttatctat cttagtcttc aaaacagccg ctgatatatc
     3301 atcaaaattc ataccctcag tttgtgttgc catttccttt agtctcaatc ttctactgat
     3361 agttgacgtt tgtgtcgaaa ctgaacttac tgatgacgat atatcagtta tttgtgttga
     3421 tacatccgtc catgctgatg ctgaagatcc aatcgaacgt atagatgaac ctcttgatat
     3481 tgatgatgct gcgtctgata aagaatctgt cagtgttgaa actgaattcg ctaatcctga
     3541 ctttttgaat tttttcataa cattagtagc cattgatttt gcagcatcaa tagtactttt
     3601 aatgccagaa aacattgaga acatatctaa tggtagaagc gctaaatcga ttaactgcga
     3661 cattgcaatt tcttgagaca aagcgttaaa ctcttctctt agttctccta actgcctttc
     3721 taaatcttgt cgtacagtaa ctgagtttgc tataggagtc tgatagtcat catttgatgg
     3781 cactaatgat ataagtgaaa atcgtccagc aatttcataa tattctttac cattatttgg
     3841 atctgcagca ggcagaccat acaatctatc taatctagtt ctagtcagtt taaagtgtgg
     3901 ttcttcgaca gctagtctaa atctaaatct taatgaattt aatgatacga aatctgtaaa
     3961 ttgagtagat agtgttacac cagctgaatg taaagaaact gctcccccag ttaaaacagg
     4021 ccattgtcca actggtagac taaaattata gctgcctcca gtacacataa ctgaattcaa
     4081 gtttgctgct aacgatcgga catacaccac gttacgaaat gcttgtgaat catcccagta
     4141 atcgatatag acgtatgaat tctctttaat tacttcaaat ttagatacaa caaaatcagt
     4201 tggtaaatat cctccattaa aactgaagtc attaacgcca ttcactgaac aagtagtatg
     4261 tgcggtaact tcttcaccat cacgagtata tgtgtattga taattcgctg gcttaaatga
     4321 tatttctgac cacttatatc ccagccctcc tgattttata attgtatttg caaatttaaa
     4381 tctaatagtt atatctctat tatattgcat ttctttccat aatgatgtct tggatatcac
     4441 aatatcttca ttagcttgag ctctatagtg tattacatct ctagcagtca acgataatgg
     4501 tacaacattt ctagtatttt gtattggtgg taatccatta ttaatgtatt ccgtacattt
     4561 agactcttca gatctaggaa ttatataaaa gtcacaaaaa gcagtcatgt taacagaatc
     4621 ataattcgtt gttgaataat gtgctgtcct agcgttaggt gtctgtcctt catatgtata
     4681 taatttttca ttatgcttca taactgcata taatttcgga ctggatagta atggtccata
     4741 ttgtcctata cttccatttg cagttgtttt tacgacatca ataaatttcc actggtcttg
     4801 tgaagtattg gataccgtta attgttcttg aataccaaat atagtgtaag ttctattttc
     4861 agactgaaca tttggctcga ttaaaattgt ggctaaccat ctatctgtat tatttgtacc
     4921 ttcaacaatt acgccaggtg tcgttggagc cagtaacatc caataatcga ctggtggatt
     4981 gaatgtcgtt ggttgataag gcccatccag caatggtcca actgtcgtag aatcattaat
     5041 ttctccaggt ccccagttaa ctggagcata acctgtttgc gcaaatggtc caggattaat
     5101 tgtgacattt tgtgatttag ttgatccaat ctcttgtatc tcatcggata aatctactgt
     5161 ataagaattc gtgagcaatt gtctataaat gagcgaagcc attttatagc ctatagtgag
     5221 tcgtattaac ccgggatcga tatccccctg cattaatgaa tcggccaacg cgcggggaga
     5281 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     5341 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5401 tcaggggata acgcaggaaa gaacatgtga gcaaaa
//
