LOCUS       sequence-176099-        5111 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Express p10-FAST in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5111)
  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 5111)
  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-FAST-p10
FEATURES             Location/Qualifiers
     source          1..5111
                     /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(762..781)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    827..882
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(881..900)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1234..1254)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1239..1568
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1419..1554
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1661..2260
                     /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(1661..1680)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2157..2177
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    2490..2624
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2527..2546)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2581..2600
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2692..2709)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2863..3451)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2943..2962)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3622..4413,4414..4482))
                     /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     4245..4264
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4483..4587)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        4618..4997
                     /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 gagcggtgat tgcgctggtc tggtttcagt gtttgggagt
      421 gttcactgtc agtcgtctaa gaataaagcc ggtggggatc tgcaagcgac ttctgttcta
      481 accacatatt ggccgcatct tgcgattggt ggcagtatca tcttgatcat cctcctgctg
      541 ggtctatttt actgttgtta tctcaagtgg aagacgtcac atattcgtcg tacttatcac
      601 aaggagttgg tggcgctaac tcgcggctat gtcaggccga tcccggcgga cgttaccagt
      661 gtctgaagat cttcgaattc actcctcagg tgcaggctgc ctatcagaag gtggtggctg
      721 gtgtggccaa tgccctggct cacaaatacc actgagatcg atctttttcc ctctgccaaa
      781 aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt
      841 attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg
      901 agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc
      961 catatgctgg ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag
     1021 ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta gatttttttt
     1081 atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta
     1141 ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta
     1201 tggagatccc tcgacctgca gcccaagctc tgtggaatgt gtgtcagtta gggtgtggaa
     1261 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     1321 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     1381 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     1441 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     1501 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     1561 tttgcaaaaa gcttgcatgc ctgcaggtcg gccgccacga ccggtgccgc caccatcccc
     1621 tgacccacgc ccctgacccc tcacaaggag acgaccttcc atgaccgagt acaagcccac
     1681 ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt
     1741 cgccgactac cccgccacgc gccacaccgt cgacccggac cgccacatcg agcgggtcac
     1801 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     1861 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     1921 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     1981 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     2041 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     2101 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     2161 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     2221 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga cgcccgcccc acgacccgca
     2281 gcgcccgacc gaaaggagcg cacgacccca tggctccgac cgaagccacc cggggcggcc
     2341 ccgccgaccc cgcacccgcc cccgaggccc accgactcta gaggatcata atcagccata
     2401 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     2461 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     2521 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     2581 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatccgctgc attaatgaat
     2641 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     2701 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     2761 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2821 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2881 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2941 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3001 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3061 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3121 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3181 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3241 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3301 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3361 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3421 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3481 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3541 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3601 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     3661 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     3721 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     3781 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     3841 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     3901 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     3961 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4021 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4081 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4141 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4201 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     4261 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     4321 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     4381 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     4441 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     4501 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     4561 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgggtcgac
     4621 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
     4681 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     4741 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     4801 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     4861 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     4921 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     4981 tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc
     5041 ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg
     5101 gggcgggggg g
//
