LOCUS       sequence-201980-         886 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  To express 6His-fSNAP protein in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 886)
  AUTHORS   Haraszti RA, Braun JE
  TITLE     Preparation of SNAP(f)-Beads for Colocalization Single-Molecule 
            Spectroscopy (CoSMoS) of RNA-Protein Complexes.
  JOURNAL   Methods Mol Biol. 2020;2113:17-22. doi: 10.1007/978-1-0716-0278-2_2.
  PUBMED    32006304
REFERENCE   2  (bases 1 to 886)
  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: "Methods Mol
            Biol."; date: "2020"; volume: "2113:17-22. doi"; pages: " 
            10.1007/978-1-0716-0278-2_2"
COMMENT     Sequence Label: pET24b-6His-fSNAP
FEATURES             Location/Qualifiers
     source          1..886
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(31..48)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(98..640)
                     /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="DKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVEV
                     PAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLKV
                     VKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLAV
                     KEWLLAHEGHRLGKPGLG"
     CDS             complement(641..658)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             669..691
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    706..730
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     complement(730..749)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(731..749)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(833..852)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 tttcggggct ttgttagcag ccggatctca gtggtggtgg tggtggtgct cgagtgcgnc
       61 cncaagcttg tcgacggagc tcgaattcgg atcctcaacc cagcccaggc ttgcccagtc
      121 tgtggccctc gtgggccagc agccactctt tcacggcgag cccgccctcg tagcccccca
      181 cgtccaggtc gccctgcacc acccggtggc aggggatcag aatgggcacg ggatttccgc
      241 tcagggcggt tttcacggcg gcggtggcgg cgggattgcc ggccagggcg gccaggtggc
      301 tgtagctgat gacctctccg aacttcacca ctttcagcag tttccacagc acctggcggg
      361 taaagctctc ctgctggaac actgggtggt gcagggctgg cacagggaac tcctcgatgg
      421 cctcaggctg gtgaaagtag gcgttgagcc aggcggtggc ctgcatcagt ggctctggtc
      481 cgcccagcac ggcggctggg gcaggcactt ccacggcgtc ggcggcagat gttcctttgc
      541 ccaggaagat gatacggtgc aggccctgtt cgcacccaga cagttccagc ttgcccagag
      601 ggctatccag ggtggtgcgc ttcatttcgc agtctttgtc gtgatggtga tggtgatgca
      661 tatgtatatc tccttcttaa agttaaacaa aattatttct agaggggaat tgttatccgc
      721 tcacaattcc cctatagtga gtcgtattaa tttcgcggga tcgagatctc gatcctctac
      781 gccggacgca tcgtggccgg catcaccggc gccacaggtg cggttgctgg cgcctatatc
      841 gccgacatca ccgatgggga agatcgggct cgccacttcg ggctca
//
