LOCUS       sequence-148820-         902 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Construct for expressing His10 - SNAPf -9E in E. Coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 902)
  AUTHORS   Su X, Ditlev JA, Hui E, Xing W, Banjade S, Okrut J, King DS, Taunton
            J, Rosen MK, Vale RD
  TITLE     Phase separation of signaling molecules promotes T cell receptor 
            signal transduction.
  JOURNAL   Science. 2016 Apr 29;352(6285):595-9. doi: 10.1126/science.aad9964. 
            Epub 2016 Apr 7.
  PUBMED    27056844
REFERENCE   2  (bases 1 to 902)
  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: 
            "10.1126/science.aad9964"; journalName: "Science"; date: 
            "2016-04-29- 29"; volume: "352"; issue: "6285"; pages: "595-9"
COMMENT     Sequence Label: His10-PS-SNAPf-9E (XSB526)
FEATURES             Location/Qualifiers
     source          1..902
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     32..50
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(103..120)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(176..721)
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     CDS             complement(731..754)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(761..790)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     RBS             807..829
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(860..879)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(861..879)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 caagacccgt ttagaggccc caaggggtta tgctagttat tgctcagcgg tggcagcagc
       61 caactcagct tcctttcggg ctttgttagc agccggatct cagtggtggt ggtggtggtg
      121 ctcgagtgcg gccgctcgag tcgacttact cttcttcctc ttcttcctct tcttcaccca
      181 gcccaggctt gcccagtctg tggccctcgt gggccagcag ccactctttc acggcgagcc
      241 cgccctcgta gccccccacg tccaggtcgc cctgcaccac ccggtggcag gggatcagaa
      301 tgggcacggg atttccgctc agggcggttt tcacggcggc ggtggcggcg ggattgccgg
      361 ccagggcggc caggtggctg tagctgatga cctctccgaa cttcaccact ttcagcagtt
      421 tccacagcac ctggcgggta aagctctcct gctggaacac tgggtggtgc agggctggca
      481 cagggaactc ctcgatggcc tcaggctggt gaaagtaggc gttgagccag gcggtggcct
      541 gcatcagtgg ctctggtccg cccagcacgg cggctggggc aggcacttcc acggcgtcgg
      601 cggcagatgt tcctttgccc aggaagatga tacggtgcag gccctgttcg cacccagaca
      661 gttccagctt gcccagaggg ctatccaggg tggtgcgctt catttcgcag tctttgtcca
      721 tggatcccag gggcccctgg aacagaactt ccagagaacc gtgatggtga tggtgatggt
      781 gatggtgatg ttcttccata tgtatatctc cttcttaaag ttaaacaaaa ttatttctag
      841 agggaaaccg ttgtggtctc cctatagtga gtcgtattaa tttcgcggga tcgagatctc
      901 gg
//
