LOCUS       sequence-91422-e        3948 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses His-tagged 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 3948)
  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 3948)
  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-6His
FEATURES             Location/Qualifiers
     source          1..3948
                     /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"
     CDS             1177..1194
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1238..1285
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1364..1819
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1451..1470)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1661..1682
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1886..1905)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2005..2027
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2065..2083)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2151..2255
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2256..2324,2325..3116)
                     /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(2474..2493)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3287..3875
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3776..3795
                     /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 tggagaagct tgcggccggt ggtggccatc atcatcatca tcattgagag
     1201 cggccgcaaa aaaaaaaaaa aaaaaaaaaa aaaaaaacta gcataacccc ttggggcctc
     1261 taaacgggtc ttgaggggtt ttttggatcc gggctggcgt aatagcgaag aggcccgcac
     1321 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc ctgtagcggc
     1381 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     1441 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     1501 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     1561 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     1621 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     1681 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     1741 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     1801 atattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     1861 cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     1921 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     1981 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     2041 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     2101 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     2161 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     2221 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     2281 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     2341 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     2401 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     2461 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     2521 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     2581 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     2641 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     2701 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     2761 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     2821 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     2881 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     2941 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     3001 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     3061 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     3121 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     3181 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     3241 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     3301 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3361 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3421 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3481 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3541 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3601 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3661 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     3721 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     3781 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     3841 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     3901 ggttcctggc cttttgctgg ccttttgctc acatggctcg acagatct
//
