LOCUS       sequence-175304-        6376 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Rotavirus SA11 VP1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6376)
  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 6376)
  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-VP1SA11
FEATURES             Location/Qualifiers
     source          1..6376
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(22..44)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     144..163
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     373..392
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        682..700
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     682..699
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      complement(768..815)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     811..829
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     promoter        complement(4276..4294)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4379..4396)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4550..5138)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4630..4649)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5309..6100,6101..6169))
                     /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     5932..5951
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6170..6274)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6342..6360
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
       61 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
      121 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccattc
      181 gacgctctcc cttatgcgac tcctgcatta agaagcagcc cagtagtagg ttgaggccgt
      241 tgagcaccgc cgccgcaagg aatggtgcaa ggagatggcg cccaacagtc ccccggccac
      301 ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg
      361 atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt
      421 gatgccggcc acgatgcgtc cggcgtagag gatctggcta gcgatgaccc tgctgattgg
      481 ttcgctgacc atttccgggt gcgggacggc gttaccagaa actcagaagg ttcgtccaac
      541 caaaccgact ctgacggcag tttacgagag agatgatagg gtctgcttca gtaagccaga
      601 tgctacacaa ttaggcttgt acatattgtc gttagaacgc ggctacaatt aatacataac
      661 cttatgtatc atacacatac gatttaggtg acactataga atacaagctt gcatgcctgc
      721 aggtcgactc tagaggatct cgatccggat atagttcctc ctttcagcaa aaaacccctc
      781 aagacccgtt tagaggcccc aaggggttat gctagttatt gctcagcggt ggcagcagcc
      841 aactcagctt cctttcgggc tttgttagca gccggatcct tttttttgag ctctccctta
      901 gccatccgag tggacgacgt cctccttcgg atgcccaggt cggaccgcga ggaggtggag
      961 atgccatgcc gacccggtca catctaagcg ttctaatctt gaaagaagtt cgcgttagtg
     1021 tacggcgaac gtaatgacgt gatgttccac atcttcttcc ataattttat catttctgat
     1081 ttagggtaat tgcaaaatag gcttatccat gaaccatttt ttatagcgta gtttataact
     1141 tctagctttg catataagtg taatatgaat gtaacagctg gtatttttcc cttaaatggt
     1201 attcttatca gcttctccaa gtccggtgaa ttgaaatcaa ataattgata gcatccataa
     1261 ttaatggaca ataaattgta cacgtatgat tctagtattc tatacttatc ttgagaataa
     1321 atctttgatc caacatacgt gtcagcgtct attttcggta ttcctaatga aatcagatat
     1381 aattgtattt cgttttcatg caatgaaatc actttataca attcttcaat tgatggctta
     1441 taaactgaat actttgatat tagtctagat atggctgact tcgaaccgtc atcttcaatt
     1501 tgataagttc tggatcctat acattgaatt atatactgta cgttatctgg caaagttggc
     1561 ataaattttt ggtattgtat aggtaagtgt gcatcactta ctgtaaaaaa tggttttata
     1621 tctctgagta tgtcatttat tgttatttta cttgatttga aggtgaccgg tttctgcaac
     1681 atagtcaata aagctgatat ctgtgcacgt cttttctcaa gcgatattgg agcgtatgaa
     1741 ttcaatttcg ctttttcagt caaagctatt cctctggaca ctatattatt ctttgaaaat
     1801 aataactgtt cagatagtct cgtgacgtaa tttttaaatg ttgatgatgc cgctatctca
     1861 tcggctattc ctgatttctg acttgataaa gacatgaacg ctctttgtat atatacttcg
     1921 tcatcagtcg ttccgtactc tataataaaa tcctccgagt caaatacttt aaatgttgaa
     1981 ttcgtagtta atacgcgttc agaaggaaat agtcttaatg atccagtaat tgcgactgac
     2041 gtcatctgca ttatcttagt taaaataaac tccctatcag tttcaaatcc tctaagtcta
     2101 tttacaatat aatttgaata taaaattgct gcttgatccc actgcgtact ctgccctctc
     2161 ttttcattat taagtagatt tattccagct cgaaaaaata tttttccacc tgcaatgtac
     2221 cttttagcaa tttctattcc tactgtggat accagagctt taactttagc attcatgcgt
     2281 gcataagttt ctcttacatc gttcgatacg tcttggatca tctgcttagt cacctctgta
     2341 ttaaattgta gcaccgcata gttatcatct ccatcaactc ttattatttt tgttgcgaat
     2401 gaatgtttgt tagaaatacg tgacaaaacc gttttaatca gtgccaaatt tgctattgaa
     2461 tttgctgctt tcgtttgttt ctctcctgat gctactgccc cgtattgtat tttcttaatg
     2521 acgttgccat ctggtatttg aacgtatgaa tccatgagat tgatttgtgt ttgtttgtat
     2581 aagtttaatg tctgaagaac tttagcatca ttagtcatgt tagctaatat gtccagtccc
     2641 attattattc ctttcctaaa tggctgtgta ttatgctgag acgaatccca ctgagataca
     2701 tccgtataca agaccattgt gttattagac aaaaaacgcg ttacatcgcc gtatgacaat
     2761 aattggtttg attgtgaata aaattcagcg tattctctcg tatgttttgc gtaaatcaac
     2821 attttttcaa ctacagcgtg ctgtgctatg aaatattcgt atggcagaat gaatattatt
     2881 ctagtccttc ttcctggaac atctcttctt cctaatggta ttggtttatc aacattcact
     2941 ggtggtatta taccaggcgt gtatctttcg ttagccatat catccatgac atgcatattc
     3001 tttttagttg aaaaaattgt tttcctacca aactttagct gccttgactc accattcgat
     3061 gctgacgaca ttgatagtaa accagctaac tcagaatctc gtagcaagtg atcatcatgt
     3121 ttaactggtt cttcaagcat tttaactact tcatctctta ttaacattgt atattcccga
     3181 tacatttcat cgtccacatt ttctgtatac tcagttttca attgatccag cgcagcatct
     3241 agcatttttt gctttctaaa tccaacatga aacgaccatg aatatatctt agccattaat
     3301 ggaaattttt cgatagaacg atcaactaac cagtcctgta tcatcttagg tatgtcaact
     3361 aatccagctt tcctcatatt gtcaagcatt tgatttaatt cgtcaaatgt ttgatccgga
     3421 actattgctc gtacatattt atttgaaaat tccaattcta attcttcatt tgtaataaat
     3481 gttccattaa tatccaccaa ggctaccaaa atcgacattg gtgatgacac taactttgct
     3541 agagtagatc tattagaata tgataacact attaactctt ttgctatcaa atagtcgtca
     3601 tgagagaaaa ctgacattaa catttcaatt gaagaatttg cccatgtcac taagtagtac
     3661 ggtttatctt tcattacgtc gtattcatat ctgtactttg cgttatgcct tggtacgcca
     3721 tatttattta tcgttgaggc tactatggtg aataggtcta atctcctttt ataaacttta
     3781 agtttttcag cgacatcatt ttcatgtttt tccaaccaga acatgactgc atttaaattt
     3841 gatgatagcg aagttaaaat tgctggatcc attagtgagt ccgtatattc ctccgctgtt
     3901 ggaaataatt cagatgttat tttattgttc tcataatcca attcattcac tgttaagtct
     3961 gcctccaatg gtttgccttt cgcatacttc accaattttc tttcaacagc gttatacttg
     4021 tcgtaagaat atgatagtat tgacagtaat gtggcatttt ctatgacatc attatattca
     4081 acaaacaact ttcttaacga taacccattc tttgagttct ctaaacactt ggaatgaaat
     4141 tcaatacatc tattttctaa ttcactgttg gaagagtaat atattggaat ttgaactgca
     4201 gattgtgaat tatatataaa tgatagatat tctgacaaga ttagattgta cttccccatt
     4261 gtacagcttt aatagcctat agtgagtcgt attaacccgg gatcgatatc cccctgcatt
     4321 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4381 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4441 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4501 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4561 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4621 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4681 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4741 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4801 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4861 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4921 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4981 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5041 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5101 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5161 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5221 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5281 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5341 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5401 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5461 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5521 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5581 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5641 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5701 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5761 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5821 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5881 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5941 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6001 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6061 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6121 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6181 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6241 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     6301 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     6361 cctttcgtct cgcgcg
//
