LOCUS       sequence-172788-        8248 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express vaccinia virus D1R in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8248)
  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 8248)
  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..8248
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..657
                     /label=chicken beta-actin promoter
     intron          658..1672
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1596..1618
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1680..1699
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(4393..4412)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4458..4513
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4512..4531)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4865..4885)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4870..5199
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5050..5185
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5292..5891
                     /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(5292..5311)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5788..5808
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    6121..6255
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6158..6177)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6212..6231
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6323..6340)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6494..7082)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6574..6593)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7253..8044,8045..8113))
                     /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     7876..7895
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8114..8218)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      481 tgggggcggg gggggggggg gggggcgcca ggcggggcgg ggcggggcga ggggcggggc
      541 ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct
      601 tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga
      661 gtcgctgcgt tgccttcgcc ccgtgccccg ctccgcgccg cctcgcgccg cccgccccgg
      721 ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc
      781 tgtaattagc gcttggttta atgacggctc gtttcttttc tgtggctgcg tgaaagcctt
      841 aaagggctcc gggagggccc tttgtgcggg ggggagcggc tcggggggtg cgtgcgtgtg
      901 tgtgtgcgtg gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg
      961 gcgcggcgcg gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg
     1021 tgccccgcgg tgcggggggg gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt
     1081 gggggggtga gcagggggtg tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc
     1141 tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc
     1201 ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc
     1261 gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccccggagcg ccggcggctg
     1321 tcgaggcgcg gcgagccgca accattgcct tttatggtaa tcgtgcgaga gggcgcaagg
     1381 acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct
     1501 tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct gtccgcgggg
     1561 ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg
     1621 cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg
     1681 caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattaatta agtttaaacg
     1741 gcgcgccgat atcttaagat ctatggatgc caacgtagta tcatcttcta ctattgcaac
     1801 gtatatagac gctttagcga agaatgcttc agaattagaa cagaggtcta ccgcatacga
     1861 aataaataat gaattggaac tagtatttat taagccgcca ttaattactt tgacaaatgt
     1921 agtgaatatc tctacgattc aggaatcgtt tattcgattt accgttacta ataaggaagg
     1981 tgttaaaatt agaactaaga ttccattatc taaggtacat ggtctagatg taaaaaatgt
     2041 acagttagta gatgctatag ataacatagt ttgggaaaag aaatcattag tgacggaaaa
     2101 tcgtcttcac aaagaatgct tgttgagact atcgacagag gaacgttata tatttttgga
     2161 ttacaagaaa tatggatcct ctatccgact agaattagtc aatcttattc aagcaaaaac
     2221 aaaaaacttt acgatagact ttaagctaaa atattttcta ggatccggtg cccagtctaa
     2281 aagttcttta ttacacgcta ttaatcatcc aaagtcaagg cctaatacat ctctggaaat
     2341 agaattcaca cctagagaca atgaaaaagt tccatatgat gaactaataa aggaattgac
     2401 aactctatca cgtcatatat ttatggcttc tccagagaat gtaattcttt ctccgcctat
     2461 taacgcgcct ataaaaacct ttatgttgcc taaacaagat atagtaggtt tggatctgga
     2521 aaatctatat gccgtaacta agactgacgg cattcctata actatcagag ttacatcaaa
     2581 cgggttgtat tgttatttta cacatcttgg ttatattatt agatatcctg ttaagagaat
     2641 aatagattcc gaagtagtag tctttggtga ggcagttaag gataagaact ggaccgtata
     2701 tctcattaag ctaatagagc ctgtgaatgc aatcaatgat agactagaag aaagtaagta
     2761 tgttgaatct aaactagtgg atatttgtga tcggatagta ttcaagtcaa agaaatatga
     2821 aggtccgttt actacaacta gtgaagtcgt cgatatgtta tctacatatt taccaaagca
     2881 accagaaggt gttattctgt tctattcaaa gggacctaaa tctaacattg attttaaaat
     2941 taaaaaggaa aatactatag accaaactgc aaatgtagta tttaggtaca tgtccagtga
     3001 accaattatc tttggagaat cgtctatctt tgtagagtat aagaaattta gcaacgataa
     3061 aggctttcct aaagaatatg gttctggtaa gattgtgtta tataacggcg ttaattatct
     3121 aaataatatc tattgtttgg aatatattaa tacacataat gaagtgggta ttaagtccgt
     3181 ggttgtacct attaagttta tagcagaatt cttagttaat ggagaaatac ttaaacctag
     3241 aattgataaa accatgaaat atattaactc agaagattat tatggaaatc aacataatat
     3301 catagtcgaa catttaagag atcaaagcat caaaatagga gatatcttta acgaggataa
     3361 actatcggat gtgggacatc aatacgccaa taatgataaa tttagattaa atccagaagt
     3421 tagttatttt acgaataaac gaactagagg accgttggga attttatcaa actacgtcaa
     3481 gactcttctt atttctatgt attgttccaa aacattttta gacgattcca acaaacgaaa
     3541 ggtattggcg attgattttg gaaacggtgc tgacctggaa aaatactttt atggagagat
     3601 tgcgttattg gtagcgacgg atccggatgc tgatgctata gctagaggaa atgaaagata
     3661 caacaaatta aactctggaa ttaaaaccaa gtactacaaa tttgactaca ttcaggaaac
     3721 tattcgatcc gatacatttg tctctagtgt cagagaagta ttctattttg gaaagtttaa
     3781 tatcatcgac tggcagtttg ctatccatta ttcttttcat ccgagacatt atgctaccgt
     3841 catgaataac ttatccgaac taactgcttc tggaggcaag gtattaatca ctaccatgga
     3901 cggagacaaa ttatcaaaat taacagataa aaagactttt ataattcata agaatttacc
     3961 tagtagcgaa aactatatgt ctgtagaaaa aatagctgat gatagaatag tggtatataa
     4021 tccatcaaca atgtctactc caatgactga atacattatc aaaaagaacg atatagtcag
     4081 agtgtttaac gaatacggat ttgttcttgt agataacgtt gatttcgcta caattataga
     4141 acgaagtaaa aagtttatta atggcgcatc tacaatggaa gatagaccgt ctacaaaaaa
     4201 ctttttcgaa ctaaatagag gagccattaa atgtgaaggt ttagatgtcg aagacttact
     4261 tagttactat gttgtttatg tcttttctaa gcggtaaaga tcttcgaatt cactcctcag
     4321 gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac
     4381 cactgagatc gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga
     4441 gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt
     4501 tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga
     4561 gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga acaaaggttg
     4621 gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag
     4681 aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt
     4741 aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact
     4801 actcccagtc atagctgtcc ctcttctctt atggagatcc ctcgacctgc agcccaagct
     4861 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4921 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4981 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     5041 actccgccca tcccgcccct aactccgccc agtttccgcc cttctccgcc ccatggctga
     5101 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     5161 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttgcatg cctgcaggtc
     5221 ggccgccacg accggtgccg ccaccatccc ctgacccacg cccctgaccc ctcacaagga
     5281 gacgaccttc catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc
     5341 cccgggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg
     5401 tcgacccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg
     5461 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga
     5521 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg
     5581 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc
     5641 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca
     5701 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc
     5761 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca
     5821 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc
     5881 ccggtgcctg acgcccgccc cacgacccgc agcgcccgac cgaaaggagc gcacgacccc
     5941 atggctccga ccgaagccac ccggggcggc cccgccgacc ccgcacccgc ccccgaggcc
     6001 caccgactct agaggatcat aatcagccat accacatttg tagaggtttt acttgcttta
     6061 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     6121 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     6181 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     6241 tatcatgtct ggatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6301 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6361 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6421 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6481 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6541 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6601 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6661 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     6721 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6781 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6841 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6901 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     6961 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7021 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7081 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7141 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7201 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7261 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7321 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7381 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7441 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7501 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7561 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7621 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7681 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7741 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7801 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7861 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7921 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7981 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8041 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     8101 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8161 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8221 catttccccg aaaagtgcca cctgggtc
//
