LOCUS       sequence-384909-         998 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  evolved sortase A (eSrtA) pentamutant with improved kinetics and 
            activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 998)
  AUTHORS   Chen I, Dorr BM, Liu DR
  TITLE     A general strategy for the evolution of bond-forming enzymes using 
            yeast display.
  JOURNAL   Proc Natl Acad Sci U S A. 2011 Jul 12;108(28):11399-404. doi: 
            10.1073/pnas.1101046108. Epub 2011 Jun 22.
  PUBMED    21697512
REFERENCE   2  (bases 1 to 998)
  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: 
            "10.1073/pnas.1101046108"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2011-07-12- 12"; volume: "108"; issue: "28"; pages: 
            "11399-404"
COMMENT     Sequence Label: sortase A pentamutant (eSrtA) in pET29
FEATURES             Location/Qualifiers
     source          1..998
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(24..41)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             499..521
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    536..560
                     /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(560..579)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(561..579)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(667..686)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gctttgttag cagccggatc tcagtggtgg tggtggtggt gctcgagttt gacttctgta
       61 gctacaaaga ttttacgtgt ttcccaaacg cctgtctctt cattgtaatc atcacaagta
      121 attaatgtta attgtttatc tttacctttt tgttcatcta gaacttctac agctgttggc
      181 ttaacgtttc ttatacttgt cattttatac ttacgtgttt cattaccaac tttaaagtac
      241 accatactac cttttttggc tgctttaaga tttgtaaatt gatagttcgg acggtcaatg
      301 aaagtgtgtc ctgcaattga aatattttga tcatctagtg attcattttc ttctgcaaag
      361 cttacacctc tatttaattg ttcgcgtgtt gctggtcctg gatatactgg ttctttaata
      421 tcagcatctg gaatttcaat atagcctgcc acttttgatt tatctttcgg aatttgaggt
      481 ttagcttgca tatgtatatc tccttcttaa agttaaacaa aattatttct agaggggaat
      541 tgttatccgc tcacaattcc cctatagtga gtcgtattaa tttcgcggga tcgagatcga
      601 tctcgatcct ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg
      661 ctggcgccta tatcgccgac atcaccgatg gggaagatcg ggctcgccac ttcgggctca
      721 tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt ggccggggga ctgttgggcg
      781 ccatctcctt gcatgcacca ttccttgcgg cggcggtgct caacggcctc aacctactac
      841 tgggctgctt cctaatgcan gagtcgcata agggagagcg tcgagatccc ggacaccatc
      901 gaatggcgca aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcngn
      961 tggtgaatgt gaaaccagta acgttatacg atgtcgca
//
