LOCUS       sequence-176191-        3740 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Rotavirus SA11 NSP5 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3740)
  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 3740)
  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-NSP5SA11
FEATURES             Location/Qualifiers
     source          1..3740
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             33..44
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        complement(90..108)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(192..209)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(363..951)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(443..462)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1122..1913,1914..1982))
                     /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     1745..1764
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1983..2087)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2155..2173
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2211..2233)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2333..2352
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2562..2581
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2871..2889
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2871..2888
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      complement(2957..3004)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3000..3018
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 aagacgattc attcttatat atagttgaag gaatagaagg aagactcgtc acgtcaatac
       61 tgagagacat cactgtagcg ctttaaaagc ctatagtgag tcgtattaac cgggatcgat
      121 atccccctgc attaatgaat cggccaatgc gcggggagag gcggtttgcg tattgggcgc
      181 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
      241 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
      301 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
      361 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
      421 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
      481 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
      541 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
      601 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
      661 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
      721 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
      781 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
      841 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
      901 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
      961 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     1021 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     1081 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     1141 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     1201 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     1261 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     1321 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     1381 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     1441 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     1501 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     1561 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     1621 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     1681 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     1741 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     1801 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     1861 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     1921 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     1981 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     2041 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     2101 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     2161 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     2221 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     2281 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     2341 gagcagattg tactgagagt gcaccattcg acgctctccc ttatgcgact cctgcattaa
     2401 gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcaag
     2461 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
     2521 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag
     2581 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg
     2641 atctggctag cgatgaccct gctgattggt tcgctgacca tttccgggtg cgggacggcg
     2701 ttaccagaaa ctcagaaggt tcgtccaacc aaaccgactc tgacggcagt ttacgagaga
     2761 gatgataggg tctgcttcag taagccagat gctacacaat taggcttgta catattgtcg
     2821 ttagaacgcg gctacaatta atacataacc ttatgtatca tacacatacg atttaggtga
     2881 cactatagaa tacaagcttg catgcctgca ggtcgactct agaggatctc gatccggata
     2941 tagttcctcc tttcagcaaa aaacccctca agacccgttt agaggcccca aggggttatg
     3001 ctagttattg ctcagcggtg gcagcagcca actcagcttc ctttcgggct ttgttagcag
     3061 ccggatcctt ttttttgagc tctcccttag ccatccgagt ggacgacgtc ctccttcgga
     3121 tgcccaggtc ggaccgcgag gaggtggaga tgccatgccg acccggtcac aaaacgggag
     3181 tggggagctc cctagtgtgt tcccaggtca gacttacaaa tcttcaatca attgcattgc
     3241 gacttgtttc attctcattc ttaatgcgaa gtatttcttc ttatatttac agttcttaca
     3301 tttaccatca tcactatctg aatcatccag tacataatca tctgaatcag actctgcttc
     3361 aattcttggg tagtgcttcc tacttttatg gttttgtttt gatttctcct ttttagtatc
     3421 cgtagatatt gatattgatg agtccaaatt agcttttact tttaagcctt tatttaagga
     3481 gaaatccact tgatcgcatc cgacattact tgaaggtcgt gattgtgctg atgaatccat
     3541 agacacgcca gcgtctgcgt ttgtcttaac tgcattcgat ctaatcgaaa aactggtgag
     3601 tggatcgttt gaagcagaat cagatggtcc aatatcctct ggagacttcg acagcatgta
     3661 tttattgaat gcttctgcat ctggtgaaat gtactgttca ctcctaccaa tagattttcc
     3721 agaaagagtt gacgttgttg
//
