LOCUS       sequence-176100-        5766 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Rotavirus SA11 VP2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5766)
  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 5766)
  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-VP2SA11
FEATURES             Location/Qualifiers
     source          1..5766
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(19..36)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(190..778)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(270..289)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(949..1740,1741..1809))
                     /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     1572..1591
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1810..1914)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1982..2000
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2038..2060)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2160..2179
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2389..2408
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2698..2716
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2698..2715
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      complement(2784..2831)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2827..2845
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     promoter        complement(5683..5701)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
       61 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
      121 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
      181 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
      241 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
      301 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
      361 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
      421 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
      481 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
      541 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
      601 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
      661 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
      721 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
      781 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
      841 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
      901 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
      961 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     1021 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     1081 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     1141 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     1201 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     1261 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     1321 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     1381 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     1441 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     1501 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     1561 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     1621 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     1681 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     1741 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     1801 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     1861 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     1921 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     1981 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     2041 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     2101 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     2161 ggcatcagag cagattgtac tgagagtgca ccattcgacg ctctccctta tgcgactcct
     2221 gcattaagaa gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg
     2281 gtgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc
     2341 gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
     2401 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
     2461 gtagaggatc tggctagcga tgaccctgct gattggttcg ctgaccattt ccgggtgcgg
     2521 gacggcgtta ccagaaactc agaaggttcg tccaaccaaa ccgactctga cggcagttta
     2581 cgagagagat gatagggtct gcttcagtaa gccagatgct acacaattag gcttgtacat
     2641 attgtcgtta gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt
     2701 taggtgacac tatagaatac aagcttgcat gcctgcaggt cgactctaga ggatctcgat
     2761 ccggatatag ttcctccttt cagcaaaaaa cccctcaaga cccgtttaga ggccccaagg
     2821 ggttatgcta gttattgctc agcggtggca gcagccaact cagcttcctt tcgggctttg
     2881 ttagcagccg gatccttttt tttgagctct cccttagcca tccgagtgga cgacgtcctc
     2941 cttcggatgc ccaggtcgga ccgcgaggag gtggagatgc catgccgacc cggtcatatc
     3001 tccacaatgg ggttggcgtt tacagttcgt tcatgatgcg catattatca aaagcaacag
     3061 cattaattgg ttcaacagta tcagctgaaa cgaacgcaag caaatctgaa tatacggtaa
     3121 atgttaggtt agacgttaac atatgcattg acgctctaaa atcaaattgt tgtggaactt
     3181 gtttatacac ttttgtagta gatgtaggaa tccaatcata attagccacc aaataaaagt
     3241 cattagaatc tgaatttatc ttatatagga tgggttttaa cgtgtcgacc tttctaagtt
     3301 tgacaatctg tgcaaaaacg gttgcatcta gtttagctat taacgaaatc actgatgaat
     3361 ccgttataaa tggtagcgct ccaactaaag ctactggttg attattaagt agcatgttag
     3421 taatttgagc ataatctcct gatctcatca attcttcaag atttatttgt tgaaatccgt
     3481 ccaagtttct ggcaatattg acgtaaccat acatctcatc tcgttctaac tgcatatctc
     3541 ggtatgctat tataactcct tgtgcaattt tatctgatgc acgttcaatc tgttccatat
     3601 tcattgctat caaattaaaa atatctaatc ttcttatttc aactggtaat agtcttaatc
     3661 tatctctcaa tctatacatt tggtcatctg gtactctcgc aacatcaaat atctgtaatc
     3721 ttttcagaaa gtcctcaact attgatttca ttttcttttg atataaattt aatctatttg
     3781 ccgcagttat aattgcaact gcgtcattaa ttctttcatt ataatttgaa tgaaaattga
     3841 cattcacatt atagtaatgg aacaatgttt gcggactcgg tataaccgta gcatttccaa
     3901 ttagcataca taatgatgtt actgatgtta attgcaattt ttcagttgtt aatgtttgaa
     3961 catgctgcat attcattgtt atgcatgcca ttaatgtctc ataattgtat gataacaatc
     4021 ttgttaaatc gacaagctga ccaagtctgt tagaaagcag caaaattcct ctctgtatag
     4081 atcttttata atcaactggc attgtgggaa actgttgtct agataattgc atcagagctt
     4141 ccattaattg gttgactaca tgaccatttc ttatattatc attcaagact tgatttaaca
     4201 ctccatcaat cactacttgt ctaaactgat tataattaac aaaatgtaac caattagcta
     4261 cctgaaaatt ttgaatctgt tgctctgcaa tttgaaaggg agtctgtgga tcaccattcc
     4321 tataatgcat tctttgcatt ccgaatgccg gataaacaat ggtatttatt atggctagtt
     4381 gacatgctac taatgattct cttataaaca tatcatttgg aatcacagtg agtaaccaca
     4441 tgcctgaaat aagtgacatg taatttgtcg ttacgaaatc taatgacatg gttctctgac
     4501 tcaacatagc agcaatcaaa gttttaaaac aatcattagc ggcttgtgag tttatacctg
     4561 ttaaaaactg agtctctgat tgtattgtca aagcttccaa ttgcaaatct tgtgacattt
     4621 tctggatttg tgcctcagta gatactaatt cttttaggtc tggcaccaca gatcttgcta
     4681 aaatataatt agatgtagtt atagtatccc agagtgactc aaaattatca tgtaaattta
     4741 acctatcttg tagcaagttt ggtctgatat atctagcagt agacggtaaa ttcctgtcca
     4801 tatttaatat atagttgaca tcatttctta ttctctctgg tatgtaattg aaaatgatat
     4861 cattatttaa tggttctacc aactgatgtt gtaagaagta ttcgttaaac gcatggtcaa
     4921 ttggatgcaa tatagatgga taattgacta catttcgatc agcttgaact cgttgtctca
     4981 tctcagctat gaatcttctt actgcacctt cggtttcttc atcttgaaaa atagcatcgc
     5041 atattgcggc tgtttcagaa tcaactactt tgccagcatc ccttgaattt ttattctcta
     5101 cagccatatc tttaagtaat agataatccg gcatttccat taatacttga tcatataaat
     5161 ttaaaaaata ctctctaaca tcataatctc catcaggaag agtatctcgt ttcaatttcc
     5221 aataccatct attacgaagc tctctttctc cattagctct ataaataggc aattgtttcg
     5281 gttcaaatat tcgaaacagt ttagtttgtt tctttgcttg ttctggtttt atgtcttcta
     5341 attttttgag tattgactct tttggttcaa atgtagggat agttttttgt aatatttcat
     5401 actgaacttc tttttgatgt tcctcttttg ttttaagtac ttctaacaac tgtttcgatt
     5461 cttctttatt agattttttt acctcatcag atatcttaac ttcttcttga ttatctgtaa
     5521 ttatctcttc tttcttagat aatacttttt ctgataattg tgatttagga ctgtcattat
     5581 taatgttctt gctatcttct ttttcttgca ttcgttcatc ttgttttaga ttcgtctcac
     5641 gacgcgctcc acgttttcga tacgccattg agcctttaat agcctatagt gagtcgtatt
     5701 aaccgggatc gatatccccc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     5761 gcgtat
//
