LOCUS       sequence-150520-         992 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses SpyTagged GlpG protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 992)
  AUTHORS   Min D, Arbing MA, Jefferson RE, Bowie JU
  TITLE     A simple DNA handle attachment method for single molecule mechanical
            manipulation experiments.
  JOURNAL   Protein Sci. 2016 Aug;25(8):1535-44. doi: 10.1002/pro.2952. Epub 
            2016 Jun 6.
  PUBMED    27222403
REFERENCE   2  (bases 1 to 992)
  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: "10.1002/pro.2952"; 
            journalName: "Protein Sci"; date: "2016-08"; volume: "25"; issue: 
            "8"; pages: "1535-44"
COMMENT     Sequence Label: pSpyTag-GlpG
FEATURES             Location/Qualifiers
     source          1..992
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(10..48)
                     /codon_start=1
                     /product="peptide derived from the CnaB2 adhesin domain of 
                     Streptococcus pyogenes FbaB"
                     /label=SpyTag
                     /note="forms a covalent isopeptide bond with the SpyCatcher
                     protein (Zakeri et al., 2012)"
                     /translation="AHIVMVDAYKPTK"
     CDS             complement(661..699)
                     /codon_start=1
                     /product="peptide derived from the CnaB2 adhesin domain of 
                     Streptococcus pyogenes FbaB"
                     /label=SpyTag
                     /note="forms a covalent isopeptide bond with the SpyCatcher
                     protein (Zakeri et al., 2012)"
                     /translation="AHIVMVDAYKPTK"
     CDS             complement(706..723)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(733..759)
                     /codon_start=1
                     /product="synthetic cistron containing a ribosome binding 
                     site (Shine-Dalgarno sequence), for enhancing the bacterial
                     expression of a downstream cistron (Schoner, 1997)"
                     /label=minicistron
                     /note="This first cistron should terminate 3 bp upstream of
                     the ATG for the second cistron."
                     /translation="MYRLNKEE"
     misc_feature    810..879
                     /label=rrnG antiterminator
                     /note="antiterminator from the E. coli rrnG leader region 
                     (Berg et al., 1989)"
     primer_bind     complement(885..907)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    896..912
                     /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)."
     promoter        complement(join(920..926,927..943,944..949))
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
ORIGIN
        1 agcttactcc ttcgtcggct tgtaggcgtc caccatcacg atgtgggcac cactttcacc
       61 actaccacat tttcgttttc gcgcattgag cgaatcaaca aaagccatcg ctaaacccac
      121 ggctaacccg gcgatgtgtg ctccgttcgc catcgacatc ccaaacaaat caaaccatcc
      181 ggcgacaatc cagatcagcg caaagataat taacccacgt tgcaggtaaa tgccactttg
      241 cggatcgcgt tcgccacgta gccagacgta gcccatcagc gcatacacca cgccagaaag
      301 cccgccaaac cacggcccgc tgaatttttg ctgcacatag ccgcttaaca gggcgctgat
      361 aagcgtaatg acaattagct taccgctacc gaggcgtttt tccaccgcac cgccgagata
      421 ccaccaccag agcaggttaa agaggatatg catcagcgag aagtgcatta acgcgtgggt
      481 gaagtaacgc cagaactcaa atttcagtgt tggatcgaat ggccaggcca gccataacat
      541 cacttcctga tcgccgagaa tttgcatggc aataaacacc accacggcgg cgatcatcat
      601 cacccaggtt accggacctg cgcgttcacg caaacaggcg gcgcctgact ctcctgaacc
      661 tttcgtcggt ttatacgcat cgaccattac aatatgagcg agctcatgat gatgatgatg
      721 atgccccatg gtttattcct ccttatttaa tcgatacatt aatatatacc tctttaattt
      781 ttaataataa agttaatcga taattccggt cgagtgccca cacagattgt ctgataaatt
      841 gttaaagagc agtgccgctt cgctttttct cagcggcgct gtttcctgtg tgaaattgtt
      901 atccgctcac aattccacac attatacgag ccggatgatt aattgtcaac agctcatttc
      961 agaatatttg ccagaaccgt tatgatgtcg gc
//
