LOCUS       sequence-79516-e         869 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  expression plasmid for N-terminal his tagged S.pyogenes SrtA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 869)
  AUTHORS   Guimaraes CP, Witte MD, Theile CS, Bozkurt G, Kundrat L, Blom AE, 
            Ploegh HL
  TITLE     Site-specific C-terminal and internal loop labeling of proteins 
            using sortase-mediated reactions.
  JOURNAL   Nat Protoc. 2013 Sep;8(9):1787-99. doi: 10.1038/nprot.2013.101. Epub
            2013 Aug 29.
  PUBMED    23989673
REFERENCE   2  (bases 1 to 869)
  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: 
            "10.1038/nprot.2013.101"; journalName: "Nat Protoc"; date: 
            "2013-09"; volume: "8"; issue: "9"; pages: "1787-99"
COMMENT     Sequence Label: pET28a-SpySrtA
FEATURES             Location/Qualifiers
     source          1..869
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(24..41)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(555..572)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(582..599)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             618..640
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    655..679
                     /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(679..698)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(680..698)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(782..801)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gctttgttag cagccggatc tcagtggtgg tggtggtggt gctcgagcta ggtagatact
       61 tggttataag aatgattaaa agctttcaat acatcggcgg gcgctttatc aaagtcgtat
      121 tctgttttta gttctccttt gacaataata cgttctgttg cttcgatatc tgtacaagtc
      181 actaaagtca cttctttgag accagctgta tcatcgataa catcaacgcg ttcaggagct
      241 accgtgaaaa catcttttat gatgtattcg taaatttttt ctttatctgt taaatagatg
      301 gacatcccat tttgtgctct ttcaagcggc gaaaagagca tttgagatga acctgtaatt
      361 ccaaaaatat gatgactggc aagagaataa ttattttctc ctcccataac ttgttcttct
      421 ttcatcgttc ctgcgccata aataagctca gtatttccta aacctttaaa aattggtaaa
      481 ttaatgccaa gctctggtat ggcaatgccc cctataacag gaagttgctg agccgccatt
      541 tgtgcttgca agacggatcc gcgcggcacc aggccgctgc tgtgatgatg atgatgatgg
      601 ctgctgcgca tggtatatct ccttcttaaa gttaaacaaa attatttcta gaggggaatt
      661 gttatccgct cacaattccc ctatagtgag tcgtattaat ttcgcgggat cgagatctcg
      721 atcctctacg ccggacgcat cgtggccggc atcaccggcg ccacaggtgc ggttgctggc
      781 gcctatatcg ccgacatcac cgatggggaa gatcgggctc gccacttcgg gctcatgagc
      841 gcttgtttcg gcgtgggtat ggtggcagg
//
