LOCUS       sequence-176192-        7290 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Express vaccinia virus D1R in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7290)
  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 7290)
  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: pCAG-D1R
FEATURES             Location/Qualifiers
     source          1..7290
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     212..234
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     296..315
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(3009..3028)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3074..3129
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3128..3147)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3481..3501)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3486..3815
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3666..3801
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3908..4507
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3908..3927)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4404..4424
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4737..4871
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4774..4793)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4828..4847
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4939..4956)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5110..5698)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5190..5209)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5869..6660,6661..6729))
                     /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     6492..6511
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6730..6834)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6865..7244
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac cccctctagc
       61 gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt
      121 gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc gcggggggac
      181 ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc
      241 tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac
      301 gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat taattaagtt taaacggcgc
      361 gccgatatct taagatctat ggatgccaac gtagtatcat cttctactat tgcaacgtat
      421 atagacgctt tagcgaagaa tgcttcagaa ttagaacaga ggtctaccgc atacgaaata
      481 aataatgaat tggaactagt atttattaag ccgccattaa ttactttgac aaatgtagtg
      541 aatatctcta cgattcagga atcgtttatt cgatttaccg ttactaataa ggaaggtgtt
      601 aaaattagaa ctaagattcc attatctaag gtacatggtc tagatgtaaa aaatgtacag
      661 ttagtagatg ctatagataa catagtttgg gaaaagaaat cattagtgac ggaaaatcgt
      721 cttcacaaag aatgcttgtt gagactatcg acagaggaac gttatatatt tttggattac
      781 aagaaatatg gatcctctat ccgactagaa ttagtcaatc ttattcaagc aaaaacaaaa
      841 aactttacga tagactttaa gctaaaatat tttctaggat ccggtgccca gtctaaaagt
      901 tctttattac acgctattaa tcatccaaag tcaaggccta atacatctct ggaaatagaa
      961 ttcacaccta gagacaatga aaaagttcca tatgatgaac taataaagga attgacaact
     1021 ctatcacgtc atatatttat ggcttctcca gagaatgtaa ttctttctcc gcctattaac
     1081 gcgcctataa aaacctttat gttgcctaaa caagatatag taggtttgga tctggaaaat
     1141 ctatatgccg taactaagac tgacggcatt cctataacta tcagagttac atcaaacggg
     1201 ttgtattgtt attttacaca tcttggttat attattagat atcctgttaa gagaataata
     1261 gattccgaag tagtagtctt tggtgaggca gttaaggata agaactggac cgtatatctc
     1321 attaagctaa tagagcctgt gaatgcaatc aatgatagac tagaagaaag taagtatgtt
     1381 gaatctaaac tagtggatat ttgtgatcgg atagtattca agtcaaagaa atatgaaggt
     1441 ccgtttacta caactagtga agtcgtcgat atgttatcta catatttacc aaagcaacca
     1501 gaaggtgtta ttctgttcta ttcaaaggga cctaaatcta acattgattt taaaattaaa
     1561 aaggaaaata ctatagacca aactgcaaat gtagtattta ggtacatgtc cagtgaacca
     1621 attatctttg gagaatcgtc tatctttgta gagtataaga aatttagcaa cgataaaggc
     1681 tttcctaaag aatatggttc tggtaagatt gtgttatata acggcgttaa ttatctaaat
     1741 aatatctatt gtttggaata tattaataca cataatgaag tgggtattaa gtccgtggtt
     1801 gtacctatta agtttatagc agaattctta gttaatggag aaatacttaa acctagaatt
     1861 gataaaacca tgaaatatat taactcagaa gattattatg gaaatcaaca taatatcata
     1921 gtcgaacatt taagagatca aagcatcaaa ataggagata tctttaacga ggataaacta
     1981 tcggatgtgg gacatcaata cgccaataat gataaattta gattaaatcc agaagttagt
     2041 tattttacga ataaacgaac tagaggaccg ttgggaattt tatcaaacta cgtcaagact
     2101 cttcttattt ctatgtattg ttccaaaaca tttttagacg attccaacaa acgaaaggta
     2161 ttggcgattg attttggaaa cggtgctgac ctggaaaaat acttttatgg agagattgcg
     2221 ttattggtag cgacggatcc ggatgctgat gctatagcta gaggaaatga aagatacaac
     2281 aaattaaact ctggaattaa aaccaagtac tacaaatttg actacattca ggaaactatt
     2341 cgatccgata catttgtctc tagtgtcaga gaagtattct attttggaaa gtttaatatc
     2401 atcgactggc agtttgctat ccattattct tttcatccga gacattatgc taccgtcatg
     2461 aataacttat ccgaactaac tgcttctgga ggcaaggtat taatcactac catggacgga
     2521 gacaaattat caaaattaac agataaaaag acttttataa ttcataagaa tttacctagt
     2581 agcgaaaact atatgtctgt agaaaaaata gctgatgata gaatagtggt atataatcca
     2641 tcaacaatgt ctactccaat gactgaatac attatcaaaa agaacgatat agtcagagtg
     2701 tttaacgaat acggatttgt tcttgtagat aacgttgatt tcgctacaat tatagaacga
     2761 agtaaaaagt ttattaatgg cgcatctaca atggaagata gaccgtctac aaaaaacttt
     2821 ttcgaactaa atagaggagc cattaaatgt gaaggtttag atgtcgaaga cttacttagt
     2881 tactatgttg tttatgtctt ttctaagcgg taaagatctt cgaattcact cctcaggtgc
     2941 aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact
     3001 gagatcgatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat
     3061 ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg
     3121 tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat
     3181 ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta
     3241 taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa
     3301 gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca
     3361 tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc
     3421 ccagtcatag ctgtccctct tctcttatgg agatccctcg acctgcagcc caagctctgt
     3481 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3541 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3601 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3661 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3721 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     3781 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct tgcatgcctg caggtcggcc
     3841 gccacgaccg gtgccgccac catcccctga cccacgcccc tgacccctca caaggagacg
     3901 accttccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtcccccg
     3961 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga
     4021 cccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg
     4081 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac
     4141 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt
     4201 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc
     4261 caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg
     4321 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc
     4381 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt
     4441 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg
     4501 tgcctgacgc ccgccccacg acccgcagcg cccgaccgaa aggagcgcac gaccccatgg
     4561 ctccgaccga agccacccgg ggcggccccg ccgaccccgc acccgccccc gaggcccacc
     4621 gactctagag gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     4681 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     4741 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4801 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     4861 atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4921 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4981 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5041 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5101 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5161 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5221 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5281 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5341 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5401 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5461 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5521 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     5581 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5641 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5701 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5761 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5821 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     5881 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5941 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6001 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6061 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6121 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6181 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6241 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6301 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6361 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6421 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6481 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6541 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6601 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6661 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6721 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6781 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6841 tccccgaaaa gtgccacctg ggtcgacatt gattattgac tagttattaa tagtaatcaa
     6901 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     6961 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     7021 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     7081 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     7141 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     7201 ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca
     7261 cgttctgctt cactctcccc atctcccccc
//
