LOCUS       sequence-443929-        5406 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express separately the Rotavirus SA11 NSP3 protein and the Rotavirus
            SA11 NSP5 fused with SNAP-tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5406)
  TITLE     Rotavirus Nonstructural Protein (NSP) plasmids
REFERENCE   2  (bases 1 to 5406)
  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"
COMMENT     Sequence Label: pT7-NSP3SA11-P2A-NSP5(wt)-SNAP
FEATURES             Location/Qualifiers
     source          1..5406
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(1..48)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     44..62
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(344..889)
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     CDS             complement(1514..1570)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     promoter        complement(2539..2557)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2642..2659)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2813..3401)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2893..2912)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3572..4363,4364..4432))
                     /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     4195..4214
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4433..4537)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4605..4623
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4661..4683)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4783..4802
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5012..5031
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5321..5339
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5321..5338
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctagtt attgctcagc
       61 ggtggcagca gccaactcag cttcctttcg ggctttgtta gcagccggat cctttttttt
      121 gagctctccc ttagccatcc gagtggacga cgtcctcctt cggatgccca ggtcggaccg
      181 cgaggaggtg gagatgccat gccgacccgg ccacataacg cccctatagc catttaggtt
      241 tttgacagtg ttagctttta actattgtgc tcatagaggg ttatgcatag atgacaacat
      301 actaacactg tatttgattg ttcaacatgt atcaaaatgg ttaacccagc ccaggcttgc
      361 ccagtctgtg gccctcgtgg gccagcagcc actctttcac ggcgagcccg ccctcgtagc
      421 cccccacgtc caggtcgccc tgcaccaccc ggtggcaggg gatcagaatg ggcacgggat
      481 ttccgctcag ggcggttttc acggcggcgg tggcggcggg attgccggcc agggcggcca
      541 ggtggctgta gctgatgacc tctccgaact tcaccacttt cagcagtttc cacagcacct
      601 ggcgggtaaa gctctcctgc tggaacactg ggtggtgcag ggctggcaca gggaactcct
      661 cgatggcctc aggctggtga aagtaggcgt tgagccaggc ggtggcctgc atcagtggct
      721 ctggtccgcc cagcacggcg gctggggcag gcacttccac ggcgtcggcg gcagatgttc
      781 ctttgcccag gaagatgata cggtgcaggc cctgttcgca cccagacagt tccagcttgc
      841 ccagagggct atccagggtg gtgcgcttca tttcgcagtc tttgtccatg gatccacttc
      901 cagaaccgga tccagaacct aaatcttcta taagttgcat ggcaacttgt ttcattctca
      961 tccttagtgc gaaatatttc ttcttatatt tacagttttt acattttcca tcatctgaat
     1021 cgctatcgtc taatacataa tcgtcagaat cactttctgc ttctattctt ggataatgtt
     1081 tcctactttt atggttttgt tttgatttct ctttttttgt atctgtgctt atagatatac
     1141 tactatctaa attggctttt acttttaagc ctttatttaa tgaaaaatct acttgatcgc
     1201 agcctacgtt agatgaaggt cttgattgtg cagaactgtc catgcttaca cctgcatcgg
     1261 catttgtttt cactgcgtta cttcggattg aaaaacttgt taaaggatca tttgatgcgg
     1321 agtctgaggg tccgatatct tcgggggatt tggataacat gtatttattg aacgcttcgg
     1381 cgtctgggct tatatattgt tcacttcttc ctatactttt tccacttagg gtacttgttg
     1441 tactactact ttcattctta tatattgtac taggtatgga aggtaaggaa gtaacgtcta
     1501 tactaagtga cataggaccg gggttttctt ccacgtctcc tgcttgcttt aacagagaga
     1561 agttcgttgc ttcgcaacca aatgaatatt gataattaca tctctgtatt aatcctttga
     1621 acataaggaa taatttgtca taatcagata tcaaattacg tattactgat tctaagtcat
     1681 caaaagcgtg caatggattt atcgcatcaa ttgagttaat ttgctgttca atgtctgcct
     1741 ttatttcagg gtctaattcc attgatctta aataccattc aatcgaagaa gtaagggatc
     1801 caattttatt ttgcaaatca cgttctagtt tattattgta cacttgaagc tcagctatat
     1861 gtgcttgctg ttgagagatg acattttgaa gagaatgcat attttcattc atttttcttg
     1921 ctttcaacac ccaattatta tatttttcat ttaccctgga tttcaatgat tcaaaccttt
     1981 gctccaattg ctcatagcgc gatttccagt caatggtatc cacttccatt ttttcatcca
     2041 caaatgaatc atccacttca acttcaccac gttccaattt atcacgcatc aattttgtgc
     2101 atttaataat agatgatgat tttccaggta ttctttttac actgaagcat gcgtttaaaa
     2161 ctctcatttt ttgatcaatt ccttttgatg ataacatcat tcttagtttg tttacatcct
     2221 catctaattt agctgctcta gaagtatcag caagccagtt tctatttctt atagcagatc
     2281 caaatttatt attcaaagct tgatcaatag ttattgcttt accaatcaga ttatttttaa
     2341 caccagaatc gtccatcaca aaatcaaatt tattctttac tctagaatat atatcctgat
     2401 aatcatattc tattcccata ttctcaagag ctgaggttgc agctacaact gcagcttcaa
     2461 aagaagagtt aataattgag acggccatct gttgcgtaga ctccatcttg agcatcaacc
     2521 actgaaaagc attaaatgcc tatagtgagt cgtattaacc cgggatcgat atccccctgc
     2581 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     2641 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     2701 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2761 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2821 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2881 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2941 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3001 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3061 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3121 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3181 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3241 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3301 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3361 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3421 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3481 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3541 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3601 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3661 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     3721 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3781 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     3841 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     3901 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3961 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     4021 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4081 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     4141 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     4201 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     4261 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4321 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4381 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     4441 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4501 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     4561 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     4621 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     4681 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     4741 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     4801 tactgagagt gcaccattcg acgctctccc ttatgcgact cctgcattaa gaagcagccc
     4861 agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcaag gagatggcgc
     4921 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
     4981 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     5041 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atctggctag
     5101 cgatgaccct gctgattggt tcgctgacca tttccgggtg cgggacggcg ttaccagaaa
     5161 ctcagaaggt tcgtccaacc aaaccgactc tgacggcagt ttacgagaga gatgataggg
     5221 tctgcttcag taagccagat gctacacaat taggcttgta catattgtcg ttagaacgcg
     5281 gctacaatta atacataacc ttatgtatca tacacatacg atttaggtga cactatagaa
     5341 tacaagcttg catgcctgca ggtcgactct agaggatctc gatccggata tagttcctcc
     5401 tttcag
//
