LOCUS       sequence-151212-        1002 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Construct for expressing His10 - SNAPf in E. Coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1002)
  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 1002)
  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 (XSB522)
FEATURES             Location/Qualifiers
     source          1..1002
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             19..41
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             58..87
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             94..117
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             127..672
                     /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             701..718
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(771..789)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      785..832
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(956..975)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tttccctcta gaaataattt tgtttaactt taagaaggag atatacatat ggaagaacat
       61 caccatcacc atcaccatca ccatcacggt tctctggaag ttctgttcca ggggcccctg
      121 ggatccatgg acaaagactg cgaaatgaag cgcaccaccc tggatagccc tctgggcaag
      181 ctggaactgt ctgggtgcga acagggcctg caccgtatca tcttcctggg caaaggaaca
      241 tctgccgccg acgccgtgga agtgcctgcc ccagccgccg tgctgggcgg accagagcca
      301 ctgatgcagg ccaccgcctg gctcaacgcc tactttcacc agcctgaggc catcgaggag
      361 ttccctgtgc cagccctgca ccacccagtg ttccagcagg agagctttac ccgccaggtg
      421 ctgtggaaac tgctgaaagt ggtgaagttc ggagaggtca tcagctacag ccacctggcc
      481 gccctggccg gcaatcccgc cgccaccgcc gccgtgaaaa ccgccctgag cggaaatccc
      541 gtgcccattc tgatcccctg ccaccgggtg gtgcagggcg acctggacgt ggggggctac
      601 gagggcgggc tcgccgtgaa agagtggctg ctggcccacg agggccacag actgggcaag
      661 cctgggctgg gttaagtcga ctcgagcggc cgcactcgag caccaccacc accaccactg
      721 agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
      781 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
      841 aggaactata tccggattgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
      901 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
      961 ttcgctttct tcccttcctt tctcgccacg ttcgccggct tt
//
