LOCUS       sequence-201979-         973 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 973)
  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 973)
  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..973
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             1..23
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             52..594
                     /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             644..661
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      728..775
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(900..919)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tttgtttaac tttaagaagg agatatacat atgcatcacc atcnccnnnn cgacaaagac
       61 tgcgaaatga agcgcaccac cctggatagc cctctgggca agctggaact gtctgggtgc
      121 gaacagggcc tgcaccgtat catcttcctg ggcaaaggaa catctgccgc cgacgccgtg
      181 gaagtgcctg ccccagccgc cgtgctgggc ggaccagagc cactgatgca ggccaccgcc
      241 tggctcaacg cctactttca ccagcctgag gccatcgagg agttccctgt gccagccctg
      301 caccacccag tgttccagca ggagagcttt acccgccagg tgctgtggaa actgctgaaa
      361 gtggtgaagt tcggagaggt catcagctac agccacctgg ccgccctggc cggcaatccc
      421 gccgccaccg ccgccgtgaa aaccgccctg agcggaaatc ccgtgcccat tctgatcccc
      481 tgccaccggg tggtgcaggg cgacctggac gtggggggct acgagggcgg gctcgccgtg
      541 aaagagtggc tgctggccca cgagggccac agactgggca agcctgggct gggttgagga
      601 tccgaattcg agctccgtcg acaagcttgc ggccgcactc gagcaccacc accaccacca
      661 ctgagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
      721 ncaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
      781 aggaggaact atatccggat tggcgaatgg ggacgcgccc tgtagcggcg cattaagcgc
      841 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
      901 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
      961 aaatcggggg ctc
//
