LOCUS       sequence-483556-        3923 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses B18R protein from Western vaccinia virus that binds to and
            neutralizes type I IFNs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3923)
  AUTHORS   Yoshioka N, Gros E, Li HR, Kumar S, Deacon DC, Maron C, Muotri AR, 
            Chi NC, Fu XD, Yu BD, Dowdy SF
  TITLE     Efficient generation of human iPSCs by a synthetic self-replicative 
            RNA.
  JOURNAL   Cell Stem Cell. 2013 Aug 1;13(2):246-54. doi: 
            10.1016/j.stem.2013.06.001.
  PUBMED    23910086
REFERENCE   2  (bases 1 to 3923)
  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"; doi: 
            "10.1016/j.stem.2013.06"; journalName: "Cell Stem Cell"; date: 
            "2013-08-1- 1"; volume: "13"; issue: "2"; pages: "246-54"
COMMENT     Sequence Label: pTNT-B18R
FEATURES             Location/Qualifiers
     source          1..3923
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        16..34
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        40..58
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     40..57
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      94..103
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     terminator      1197..1244
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1339..1794
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1426..1445)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1636..1657
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1861..1880)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1980..2002
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2040..2058)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2126..2230
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2231..2299,2300..3091)
                     /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     complement(2449..2468)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3262..3850
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3751..3770
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taaggctaga gtacttaata cgactcacta taggctagca tttaggtgac actatagaat
       61 acaagctact tgttcttttt gcactcgaga attgccacca tgggcacgat gaaaatgatg
      121 gtacatatat atttcgtatc attattgtta ttgctattcc acagttacgc catagacatc
      181 gaaaatgaaa tcacagaatt cttcaataaa atgagagata ctctaccagc taaagactct
      241 aaatggttga atccagcatg tatgttcgga ggcacaatga atgatatagc cgctctagga
      301 gagccattca gcgcaaagtg tcctcctatt gaagacagtc ttttatcgca cagatataaa
      361 gactatgtgg ttaaatggga aaggctagaa aaaaatagac ggcgacaggt ttctaataaa
      421 cgtgttaaac atggtgattt atggatagcc aactatacat ctaaattcag taaccgtagg
      481 tatttgtgca ccgtaactac aaagaatggt gactgtgttc agggtatagt tagatctcat
      541 attagaaaac ctccttcatg cattccaaaa acatatgaac taggtactca tgataagtat
      601 ggcatagact tatactgtgg aattctttac gcaaaacatt ataataatat aacttggtat
      661 aaagataata aggaaattaa tatcgacgac attaagtatt cacaaacggg aaaggaatta
      721 attattcata atccagagtt agaagatagc ggaagatacg actgttacgt tcattacgac
      781 gacgttagaa tcaagaatga tatcgtagta tcaagatgta aaatacttac ggttataccg
      841 tcacaagacc acaggtttaa actaatacta gatccaaaaa tcaacgtaac gataggagaa
      901 cctgccaata taacatgcac tgctgtgtca acgtcattat tgattgacga tgtactgatt
      961 gaatgggaaa atccatccgg atggcttata ggattcgatt ttgatgtata ctctgtttta
     1021 actagtagag gcggtattac cgaggcgacc ttgtactttg aaaatgttac tgaagaatat
     1081 ataggtaata catataaatg tcgtggacac aactattatt ttgaaaaaac ccttacaact
     1141 acagtagtat tggagtaagc ggccgcaaaa aaaaaaaaaa aaaaaaaaaa aaaaaactag
     1201 cataacccct tggggcctct aaacgggtct tgaggggttt tttggatccc ggcgcgccgg
     1261 gatccgggct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     1321 agcctgaatg gcgaatggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     1381 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     1441 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     1501 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     1561 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     1621 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     1681 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     1741 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttcctgat
     1801 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag
     1861 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     1921 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     1981 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     2041 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     2101 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     2161 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     2221 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     2281 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     2341 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     2401 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     2461 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     2521 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     2581 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     2641 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     2701 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     2761 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     2821 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     2881 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     2941 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     3001 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     3061 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     3121 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     3181 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3241 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3301 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3361 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     3421 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3481 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3541 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3601 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3661 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3721 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3781 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3841 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3901 tgctcacatg gctcgacaga tct
//
